Background:This study evaluated whether high-density lipoprotein (HDL) treatment, shown to preserve endothelial cell integrity, could protect the left ventricular ejection fraction (LVEF) in rats from ischemia-reperfusion (IR) injury. Methods:Human umbilical vein endothelial cells (HUVECs) were assigned to six groups: G1 (HUVECs alone); G2 (+healthy-derived oxidized LDL [ox-LDLH]); G3 (+ox-LDLH + healthy-derived HDL [HDLH]); G4 (+acute-coronary-syndrome patient-derived ox-LDL [ox-LDLP]); G5 (+ox-LDLP+ HDLH); and G6 (+ox-LDLP + patient-derived HDL [HDLP]). Male Sprague-Dawley rats (n = 54) were divided equally into Group 1 (subcutaneous control), Group 2 (IR only), and Groups 3-6 (IR plus ox-LDLH, HDLH, ox-LDLP, or HDLP, respectively, implanted into the left ventricular myocardium 3 h after IR). Results:In vitro, protein and cellular markers of apoptosis, oxidative stress, mitochondrial damage, and MAPK-family signaling were lowest in G1, highest in G4, and significantly lower in G3 than in G2, G5, and G6 (all p < 0.0001); antioxidants and angiogenesis showed the opposite pattern. By day 28, LVEF and LV fractional shortening were highest in Group 1, lowest in Group 5, higher in Group 4 than in Groups 2, 3, and 6, and higher in Group 6 than in Groups 2 and 3 (all p < 0.0001). Inflammation, MAPK cascades, oxidative stress, mitochondrial damage, and LV infarct/fibrotic areas followed the inverse pattern, while antioxidants paralleled LVEF. Conclusions:HDLH protected heart function against IR-induced myocardial damage by inhibiting inflammation and oxidative stress and modulating MAPK-family signaling.

INTRODUCTION:We tested whether early administration of multiple doses of adipose-derived mesenchymal stem cells (ADMSCs) overexpressing cellular prion protein (PrPC-OVE) could preserve lung parenchyma and function in a rodent model of bleomycin-induced pulmonary fibrosis (PF). METHODS:Cell culture, immunohistochemistry, immunofluorescence, and western blot analyses were conducted. Animals were categorized into sham-control (Group 1), PF (Group 2), PF + ADMSCs (one dose; Group 3), PF + ADMSCs (three doses; Group 4), PF + PrPC-OVE in ADMSCs (one dose; Group 5), and PF + PrPC-OVE in ADMSCs (three doses; Group 6). RESULTS:In vitro, bleomycin suppressed L2 cell proliferation and increased apoptosis and epithelial-mesenchymal transition (EMT) markers, which were significantly reversed by ADMSCs and further enhanced by PrPC-OVE in ADMSCs. TGF-β/Smads signaling-mediated EMT was identified as a crucial mechanism in cellular fibrosis and PF, which was significantly suppressed by silencing TGF-β in L2 cells and PrPC-OVE in ADMSCs co-cultures and lung tissue. By days 28 and 42 after PF induction, O₂ saturation (%) was highest in Group 1, lowest in Group 2, and notably higher in Group 6 than in Groups 3-5. Right-ventricular (RV) systolic blood pressure showed an inverse trend. RV and left lung weights-to-tibial length ratios were significantly greater in Group 2 at both time points. By day 42, Group 2 exhibited the highest lung injury, fibrosis, and protein levels of fibrotic, EMT, inflammatory, and oxidative stress markers, whereas these parameters were lowest in Group 1 and significantly higher in Group 4 than in Group 6. DISCUSSION:This study, which investigated the therapeutic role of PrPC-OVE in ADMSCs in protecting lung parenchyma against bleomycin-induced damage, provides several important preclinical insights. First, the in vitro study demonstrated that bleomycin activated TGF-β/Smads signaling, inducing EMT upregulation in lung epithelial cells (L2 cell line), which plays a crucial role in the initiation and propagation of PF. Second, both in vitro and in vivo studies showed that PrPC-OVE played a fundamental role in attenuating bleomycin-induced PF, mainly through regulation of TGF-β/Smads signaling. Third, ADMSCs were less effective than PrPC-OVE in ADMSCs, and repeated doses of ADMSCs were less effective than repeated doses of PrPC-OVE in ADMSCs in protecting lung function and parenchyma against bleomycin-induced damage. CONCLUSION:Repeated PrPC-OVE-ADMSCs effectively preserved lung function and parenchyma by suppressing TGF-β/Smads signaling in bleomycin-induced PF.

Background: This study investigated whether the double overexpression of decorin and IL-10 in human bone marrow-derived mesenchymal stem cells [HBMDMSCs DO (DEC/IL-10) ] can protect against bleomycin (BLM)-induced pulmonary fibrosis (PF) in mice. Methods: L2 and NR8383 cells were treated with BLM (0–2.5 μg/mL). In vivo , animals were assigned to four groups: Group 1 – sham control (SC); Group 2 – PF; Group 3 – PF + HBMDMSCs; Group 4 – PF + HBMDMSCs DO (DEC/IL-10) . Results: BLM, at its IC 50 , significantly reduced the viability of L2 and NR8383 cells while increasing early and late apoptosis and TGF-β expression ( P < 0.001). BLM treatment in L2 cells induced oxidative stress (mitochondrial ROS, NOXs/p22 phox ), cell-stress signaling (MAPKs), and epithelial–mesenchymal transition (EMT) biomarkers, which were reversed by decorin, HBMDMSCs, decorin-overexpressing HBMDMSCs, or p22 phox silencing ( P < 0.001). Decorin silencing in L2 and CG1629 cells upregulated EMT and cell-stress markers, effects reversed by decorin treatment, decorin overexpression, or MSC DO (DEC/IL-10) ( P < 0.001). In BLM-induced PF mice, SaO 2 was highest in Group 1, lowest in Group 2, and higher in Group 4 than Group 3, while lung injury, fibrosis, right-ventricular pressure, and inflammatory cells showed the opposite trend ( P < 0.0001). Lung expression of EMT/proliferation (c-Myc, Cyclin D/E, Wnt/β-catenin), oxidative stress (p22 phox , OxyBlot), and fibrosis (TGF-β, p-Smad3) proteins mirrored lung injury scores ( P < 0.0001). Conclusion: HBMDMSCs DO (DEC/IL-10) treatment effectively protected the lungs from BLM-induced PF in rodents.

This study tested the hypothesis that mitofusin-2 overexpression (Mfn2Ove) in adipose-derived mesenchymal stem cells (ADMSCOve−Mfn2) protects the testis against testicular torsion-induced ischemia‒reperfusion (TTIR) injury via AMP-activated protein kinase (AMPK)-mediated mitochondrial dynamic homeostasis. The in vitro results revealed that Mfn2Ove significantly increased cell viability/mitochondrial content/fusion protein/ATP levels (all P < 0.001). Mfn2Ove in ADMSCs and GC-1 cells upregulated the expression of mitochondrial fusion proteins/content/OXPHOS complexes/p-AMPK and suppressed fission proteins/autophagy/oxidative stress, which were reversed by siRNA-Mfn2 (all P < 0.001). Compared with the control, Mfn2Ove significantly increased mitochondrial length in ADMSCs and GC-1 cells (P < 0.0001). IR activated p-AMPK and upregulated fission proteins, whereas the AMPK inhibitor did not reverse the ability of Mfn2Ove to upregulate p-AMPK/Mfn2/OPA1/OXPHOS/mitochondrial length in ADMSCs and GC-1 cells (P < 0.0001). By day 28 after TTIR induction, testicular protein levels of oxidative stress/fibrosis/apoptosis/autophagy/DNA-damage/fission were significantly increased in group 2 (TTIR) compared with those of group 1 (sham-control), whereas the protein levels of mitochondrial fusion/OXPHOS exhibited an opposite pattern of oxidative stress between groups 1 and 2 that were significantly reversed by AMDCs (group 3) and further reversed by ADMSCOVe−Mfn2 (group 4), which was reversed by siRNA-Mfn2 in ADMSCs (group 5) (all P < 0.00001). The p-AMPK protein level significantly increased from groups 1 to 4 but was reversed in group 5 (all P < 0.00001). The cellular levels of androgen receptor/Sertoli cells/alpha-microtubule/vimentin/testis injury score showed identical patterns to those of OXPHOS complexes among the groups (all P < 0.00001). The activation of the AMPK–Mfn2 axis protected the testis against TTIR injury.

Background: We tested the hypothesis that febuxostat (Feb) therapy effectively protected cardiorenal syndrome (CRS) rats via repressing the xanthine-oxidase (XO)-caused oxidative stress. Methods and Results: Cellular levels of apoptosis/oxidative stress/mitochondrial-membrane potential were higher in p-Cresol treated-NRK-52E cells than in control group that were reversed by Feb treatment or silencing XO gene (all P<0.001). Pilot study demonstrated that: XO activity was significantly increased in CRS than in SC group; a significant negative correlation between XO activity and left ventricular ejection fraction (LVEF) (%); a significant positive correlation between XO activity and BNP/BUN/creatinine/proteinuria levels (all P<0.01). Male-adult SD-rats were classified into groups 1(sham-control)/2 (CRS)/3 [CRS+Feb (10mg/kg/day)]/4 [CRS+Feb (30mg/kg/day)]. By day-63, the survival rate was significantly lower in group 2 than in other groups (P=0.029), and circulatory levels of FGF23/BNP/XO-activity BUN/creatinine/proteinuria and renal-artery resistance were highest in group 2/lowest in group 1/significantly lower in group 4 than in group 3, whereas the LVEF exhibited an opposite pattern of XO among the groups (all P<0.0001). Cellular levels of fibrosis/XO/H2DCFDA/CD68/CHAC1, and protein expressions of oxidative-stress (NOX-2/NOX-4/XO)/inflammatory (NF-κB/IL-1β)/fibrotic (Smad3/TFG-β)/apoptotic (CHAC1/2)/mitochondrial-damaged (p-DRP1) biomarkers in kidney/heart tissues displayed a similar pattern of XO (all P<0.0001). Conclusion: Feb therapy improved cardiorenal function and prognostic outcome in CRS rats.

BACKGROUND:This study assessed whether alirocumab-facilitated 5-FU therapy effectively inhibited colorectal cancer (CRC) cell proliferation/growth in the livers of nude mice. METHODS AND RESULTS:In vitro results revealed that (a) cell viability at 6, 24, 48 and 72 h, (b) wound healing ability, (c) cell migratory ability, (d) protein levels of cell proliferation/antiapoptotic signalings, and (e) protein expressions of cell cyclins were significantly and progressively reduced from groups A 1 (DLD-1 cells) and A 2 (HCT116 cells), groups A 3 (siRNA-PCSK9 in DLD-1) and A 4 (siRNA-PCSK9 in HCT116), groups to A 5 (DLD-1 cells + alirocumab 10 µM) and A 6 (HCT116 cells + alirocumab 10 µM) (n = 3-6) ( all P < 0.001 ), whereas the protein expression of apoptosis exhibited contrasting effects (n = 3) ( all P < 0.001 ). These parameters, including: (a, b, c, d, and e) and colony formation units (CFUs) were significantly and progressively reduced from groups B 1 (DLD-1), B 2 (DLD-1 + alirocumab 5 µM) to B 3 (DLD-1 + alirocumab 20 µM) (n = 3-6) ( all P < 0.001 ), and also significantly and progressively from groups C 1 [PCSK9 gene overexpression in DLD-1 (PCSK9 OE in DLD-1)], C 2 (PCSK9 OE in DLD-1 + alirocumab 5 µM) to C 3 (PCSK9 OE in DLD-1 + alirocumab 20 µM) (n = 3-6) ( all P < 0.001 ). Protein expression of PCSK9 was significantly and progressively increased from groups D 1 (CCD-18Co), D 2 (DLD-1) to D 3 (HCT116), and also significantly and progressively increased from groups from D 4 (THLE-2), D 5 (Huh7) to D 6 (Hep3B) (n = 3) ( all P < 0.001 ). For the animal study (n =10 for each group), two CRC cell lines were implanted into livers of nude mice and were categorized into groups G A1 (DLD1), G A2 (HCT116), G B1 (DLD1 + alirocumab), G B2 (HCT116 + alirocumab), G C1 (DLD1 + 5FU), G C2 (HCT116 + 5FU), G D1 (DLD1 + alirocumab + 5FU), and G D2 (HCT116 + alirocumab + 5FU). By day 35, the ratio of liver weight to body weight, mean bioluminescence intensity at days 21, 28, and 35, protein expressions of cell proliferation, cell cyclins, cellular levels of PCSK9, and extracellular matrices were significantly and progressively reduced from G1, G2, and G3 to G4 (n = 6-10) ( all P < 0.0001 ). CONCLUSION:Alirocumab-facilitated 5-FU therapy effectively suppresses PCSK9-promoted CRC proliferation/growth/invasion, highlighting that alirocumab therapy may be an alternative choice of adjuvant therapy in combination with surgery or chemotherapy.

We examined whether combined exogenous mitochondria (ExMito) and cellular prion protein overexpression (Ove-PrPC) in adipose-derived mesenchymal stem cell (Ove-PrPC in ADMSCs) therapy is superior to a single therapy for protecting the brain against intracranial hemorrhage (ICH) in rats. In vitro, compared with the control group, ExMito transfusion into recipient cells (i.e., N2a cells) significantly increased under hypoxic conditions (P < 0.001) and augmented ρ0 cell proliferation and cell-cycle activation (P < 0.001). PrPC−OE in ADMSCs exhibited higher resistance to H2O2-induced cell senescence and mitochondrial and DNA damage compared to ADMSCs (P < 0.001). Rats were categorized into group 1 (sham-control), 2 (ICH), 3 [ICH + ExMito (350 μg) by intracranial injection at 3 h after ICH], 4 [ICH + PrPC−OE in ADMSCs (6.0 × 105 cells) and intracranial injection and 1.2 × 106 cells by intravenous injection)], and 5 (ICH + combined ExMito + PrPC−OE in ADMSCs). By day 28, the brain infarct volume, brain infarct area, inflammatory cell infiltration, and biomarkers for DNA and mitochondrial damage were highest in group 2, lowest in group 1, and significantly lower in group 5 than in groups 3 and 4. NeuN cells exhibited the opposite pattern for brain infarct volume, and neurological function (corner test) significantly improved in groups 3 and 4, with further improvement in group 5 compared with that in group 2 (P < 0.0001). Combined ExMito + PrPC−OE ADMSCs therapy was superior to either therapy alone in mitigating the ICH-induced brain damage.

Background and Aims: Ischemic stroke (IS) remains the third leading cause of death, and the treatment of acute ischemic stroke (AIS) is still a formidable challenge to clinicians. This study tested the hypothesis that combined silencing Mnf2 gene in adipose-derived mesenchymal stem cells (ADMSCssil-Mnf2) and melatonin (Mel) therapy was superior to monotherapy on attenuating the brain infarct volume (BIV) and improving neurological function in AIS rats. Results: In vitro and in vivo studies were conducted. In vitro results showed that as compared with the controls (i.e., ADMSCs/N2a cells), the cellular/protein levels of oxidative stress/reactive oxygen species (ROS)/mitochondrial and DNA damaged/apoptotic/cell stress signaling (tumor necrosis factor [TNF] receptor associated factor 6/ apoptosis signal regulating kinase/MKK4/7/JUN/ERK1/2/c-Jun) biomarkers were significantly increased in these cells treated by H2O2 that were significantly reversed by ADMSCssil-Mnf2 or Mel and further significantly reversed by combined therapy (all p < 0.0001). Animals were categorized into groups 1 (sham-operated control)/2 (AIS)/3 (AIS + Mel)/4 (AIS + ADMSCssil-Mnf2)/5 (AIS + Mel-ADMSCssil-Mnf2) and euthanized by day 28 after AIS. By day 28, the BIV and the brain infarct area (BIA) were lowest in group 1/highest in group 2/significantly lower in group 5 than in groups 3 and 4/significantly increased in group 4 than in group 3, whereas the neurological function displayed an opposite manner of BIV (all p < 0.0001). The protein expressions of oxidative stress/mitochondrial damaged/apoptotic/inflammatory/cell stress signaling biomarkers displayed an identical pattern, whereas the protein expressions of mitochondrial biogenesis/antioxidants and cellular level of neuronal cells exhibited an opposite manner of BIV among the groups (all p < 0.0001). Innovation and Conclusion: ADMSCssil-Mnf2 and Mel combined therapy offered synergic effects on attenuating the BIV/BIA and preserving neurological function in rodents after AIS mainly through suppressing oxidative stress/ROS/inflammatory signalings and upregulating antioxidants. Combined ADMSCssil-Mnf2 and Mel therapy offered additional benefits on protecting the brain against AIS in rodents. Antioxid. Redox Signal. 43, 427-447.

Cardiovascular diseases (CVDs), the leading global cause of mortality, underscore an urgent need for innovative diagnostic and therapeutic strategies. Extracellular vesicles (EVs)—lipid-bilayer nanoparticles transporting bioactive cargo (microRNAs, proteins, lipids)—are critical mediators of intercellular communication in CVD pathogenesis. They exhibit functional duality: propagating pathology (inflammation, fibrosis, thrombosis) while facilitating tissue repair. This review synthesizes EV biogenesis mechanisms, isolation methodologies, source-specific functions, and multifaceted roles in atherosclerosis, myocardial infarction, heart failure, and stroke. We further evaluate EV-based diagnostic biomarkers, engineered therapeutic applications, clinical translation challenges, and future directions.

This study presents an examination of whether the double overexpression of ZNF746 and cellular prion protein (PrPC) genes in rat adipose-derived mesenchymal stromal cells (ADMSCs) (ie, MSCDGe-OVE) offered enhanced protection to the livers of rats against ischemia‒reperfusion (IR) injury. The in vitro results revealed that compared with those of rat ADMSCs, cell activities (viability/proliferation/growth/cell cycle process) were significantly upregulated by the overexpression of either gene in rat ADMSCs and were further significantly increased by MSCDGe-OVE, whereas the expression of biomarkers of oxidative stress/ROS/apoptosis/fibrosis/autophagy decreased with increasing cell viability among the groups (all P < 0.001). Male adult SD rats (n = 50) were equally categorized into groups 1 (sham-operated-control), 2 (IR), 3 (IR-MSCOVE-PrPC), 4 (IR-MSCOVE- ZNF746), and 5 (IR-MSCDGe-OVE), and livers were harvested by day 3. By day 3, the number of circulatory inflammatory/immune cells, protein expression of oxidative stress/apoptotic/fibrotic/mitochondrial damage/autophagic biomarkers, and cellular levels of DNA damage/fibrosis/inflammation in the liver parenchyma were lowest in group 1, highest in group 2 and significantly lower in groups 3/4 than in group 5 (all P < 0.0001). Liver fibrosis detected by ultrasound and the liver injury score displayed identical patterns of circulatory levels of immune cells among the groups (all P < 0.0001). Upstream and downstream inflammatory and cell-stress signaling pathways were identified as playing crucial roles in acute liver IR injury. In conclusion, MSCDGe-OVE enhanced cell proliferation and growth and ameliorated IR-induced liver damage.

BACKGROUND:This study investigated whether the combined treatment of melatonin and tofacitinib offers enhanced protection against dextran sulfate sodium-induced acute colitis (AC) in rats. Using CCD-18Co fibroblasts and a rat colitis model, we assessed the anti-inflammatory, anti-apoptotic, and immunomodulatory effects of the combination therapy. METHODS:CCD-18Co cells were grouped as A1 (CCD-18Co), A2 (CCD-18Co + lipopolysaccharide (LPS)), A3 (CCD-18Co + LPS + Melatonin), A4 (CCD-18Co + LPS + Tofacitinib), or A5 (CCD-18Co + LPS + melatonin + tofacitinib). Sprague-Dawley rats were categorized into groups 1 (normal control), 2 (AC), 3 (AC + melatonin), 4 (AC + tofacitinib), and 5 (AC + melatonin + tofacitinib), and the colons were harvested 14 days after AC induction. KEY FINDINGS:Cell viability at time points of 24, 48, and 72 h was the highest in A1, lowest in A2, and progressively increased from A3 to A5 (all P < 0.0001). The protein expression levels of inflammatory, DNA-damaged, and autophagic (ratio of LC3-BII to LC3-BI) biomarkers displayed identical patterns of apoptosis among the groups (all P < 0.0001). Additionally, the blood stool, colon leakage, and colon injury scores were the lowest in group 1, highest in group 2, and significantly progressively reduced from groups 3 to 5 (all P < 0.0001). The protein expression of the Janus kinase family-signal transducer and activator of transcriptions/cell-stress signaling, inflammation, oxidative stress, DNA-damaged, apoptotic biomarkers, and cellular expression of immune and inflammatory factors exhibited an identical pattern of colon injury scores among the groups. CONCLUSIONS:Combined melatonin-tofacitinib treatment effectively protected the colon against dextran sulfate sodium-induced damage, mainly through the suppression of Janus kinase family-signal transducer and activator of transcriptions/cell-stress signaling, inflammation, and oxidative stress.

We tested how the coordination between cyclin D1/cyclin-dependent kinase (CDK) and the cellular prion protein (PrPC) activates mitogenic/cell proliferation signaling to improve neurological outcomes in acute ischemic stroke (AIS) rats. Compared with those in adipose-derived mesenchymal stem cells (ADMSCs) and the N2a cell line, the cell viability, cell proliferation, cell-stress signaling, and wound healing rates were significantly increased upon overexpression of PrPC (PrPC-OE) in ADMSCs (all P<0.001). The cell viability, proliferation. mitochondrial mass, and protein expression of mitogenic signaling markers (cyclin D1, cyclin E1, CDK2, and CDK4) were significantly increased upon PrPC-OE in ADMSCs compared to ADMSCs that were subjected to a significant reversal of PrPC-OE by treatment with promazine (a PrPC formation inhibitor) (all P<0.001). After 3 h of serum-free/hypoxic conditions, the protein expression levels of cyclin D1/CDK, p-Akt and mitogenic signaling markers were significantly increased upon PrPC-OE in ADMSCs compared with ADMSCs that were treated with palbociclib (a cyclin D1/CDK inhibitor). Adult male Sprague-Dawley rats (n=40) were grouped into Groups 1 (AC), 2 (AIS), 3 (AIS + ADMSCs), and 4 (AIS + ADMSCs with PrPC-OE). By Day 28 after AIS induction, the neurological function and numbers of NeuN+ cells and myelin basic protein (MBP)+ cells were lowest in Group 2, highest in Group 1 and significantly increased in Group 4 compared with Group 3, whereas the cellular levels of fibrosis and inflammation markers and protein levels of markers of apoptosis, mitochondrial and DNA damage and autophagy exhibited the opposite pattern to neurological function, and protein expression levels of cell-stress signaling proteins (PI3K, Akt, and m-TOR) and PrPC progressively increased from Groups 1 to 4 (all P<0.0001). In conclusion, activated cyclin D1/CDK coordinated with PrPC to improve neurological function in the AIS setting.

Background: This study investigated whether Nanog-overexpressing adipose-derived mesenchymal stem cells (NanogOE-ADMSCs) are superior to unmodified ADMSCs in improving the left ventricular ejection fraction (LEVF) in acute myocardial infarction (AMI) patients. Methods: We utilized silencing and overexpression of Nanog gene in ADMSCs and performed a wound healing assay/transwell migration assay/MTT cell viability assay/left coronary artery ligation for AMI induction. Additionally, we categorized the cells into three classes [i.e., (ADMSCs and NanogOE-ADMSCs); A1 (ADMSCs)/A2 (ADMSCs + CoCl2)/A3 (NanogOE-ADMSCs + CoCl2)/A4 (siRNA-Nanog-ADMSCs) + CoCl2); B1 (ADMSCs)/B2 (ADMSCs + H2O2)/B3 (NanogOE-ADMSCs + H2O2)/B4 (siRNA-Nanog gene in ADMSCs + H2O2)], and the rats (n=50) were evenly divided into Groups 1 (sham-operated control)/2 (AMI)/3 (AMI+ADMSCs)/4 (AMI+NanogOE-ADMSCs)/5 (AMI+siRNA-Nanog-ADMSCs). The hearts were harvested on Day 35. Results: In vitro experiments revealed significantly higher ATP, relative mitochondrial DNA/Nonog gene expression, mitochondrial cytochrome C+ cell, angiogenesis and exosome-specific marker (Alix/CD81/CD63/CD9) levels in NanogOE-ADMSCs than in ADMSCs. The cell viability, wound healing, and migration were highest in A1, lowest in A4, and significantly greater in A3 than in A2, whereas early/late apoptosis and intracellular and mitochondrial ROS displayed the opposite pattern of cell viability among the groups (all P<0.001). Additionally, the proteins expressions of phosphorylation (p) of the PI3K/Akt/mTOR, p-JAK2/p-STAT3, and Ras/Raf/MEK1/2/ERK1/2 signaling pathways were highest in A3, lowest in A4 and significantly greater in A1 than in A2 (all P<0.001). The levels of cell cycle proteins and mitochondrial electron transport train (ETC) complex I/II/III/IV components exhibited identical patterns as PI3K/Akt/mTOR among the groups B1 to B4 (all P<0.001). On Day 35, the LVEF was highest in Group 1, lowest in Group 2, significantly greater in Group 4 than in Groups 3 and 5, and significantly greater in Group 3 than in Group 5, with the opposite pattern for the LV remodeling index, infarct and fibrosis areas, and LV chamber size (all P < 0.0001). The p-AK/p-STAT3, p-PI3K/p-Akt/p-mTOR, and Ras/Raf/MEK1/2/ERK1/2 protein levels displayed the same pattern as the LVEF among the groups (all P < 0.001). Conclusion: NanogOE-ADMSCs rescued LVEF by upregulating JAK/STAT3-mediated cell proliferation/cell stress pathways and accelerating the cell cycle.

Peritoneal fibrosis (PF) is a major complication of long-term peritoneal dialysis, leading to ultrafiltration failure and technique dropout, highlighting the urgent need for therapies that can preserve peritoneal membrane function and longevity. The present study evaluated the effectiveness of dulaglutide in preserving the functional integrity and durability of the peritoneum while inhibiting PF. In vitro Met-5A cells showed significant upregulation of inflammatory, oxidative stress, intracellular and mitochondrial reactive oxygen species (ROS), fibrotic, intracellular cytoskeletal, apoptotic and epithelial-mesenchymal transition (EMT) biomarkers, and dipeptidyl peptidase 4 (DPP4), following stimulation with a uremic toxin (p-Cresol), PF inducer [chlorhexidine gluconate (CG)] or endotoxin [lipopolysaccharide (LPS)]. Notably, these effects were significantly suppressed by dulaglutide or TGF-β/DPP4 double silencing. Furthermore, cell viability and glucagon-like peptide 1 (GLP-1) expression displayed an opposite pattern to ROS among the groups. Sprague-Dawley rats were divided into the following groups: i) Sham control (SC); ii) chronic kidney disease (CKD); iii) CKD + CG (mimicking renal failure and PF); and iv) CKD + CG + dulaglutide, and were euthanized by day 42. At this time point, the highest levels of peritoneal protein expression levels of oxidative stress (NOX-1, NOX-2 and DPP4), inflammation (NF-κB and TNF-α), angiogenesis (CD31 and von Willebrand factor) and EMT (TGF-β, Snail, β-catenin, vimentin, phosphorylated-Smad3, α-smooth muscle actin, collagen I, N-cadherin and fibronectin) factors; and cellular expression levels of fibrosis and inflammation markers, were observed in the CKD + CG group, the lowest were detected in the SC group, and the levels were significantly reduced in the CKD + CG + dulaglutide group compared with those in the CKD group. Furthermore, the expression levels of antioxidant proteins (nuclear factor erythroid 2-related factor 2, NAD(P)H quinone oxidoreductase 1 and GLP-1 receptor) exhibited an opposite trend to ROS-associated proteins among the groups. Additional Sprague-Dawley rats were categorized into the following groups: i) SC; ii) LPS-induced peritonitis; iii) LPS-induced peritonitis + dulaglutide, and were euthanized by day 5 after peritonitis induction. At this time point, flow cytometry revealed significantly increased levels of inflammatory cells (CD11b/c+, myeloperoxidase+ and Ly6G+ cells) in the circulation and abdominal fluid, and increased peritoneal permeability in the LPS-induced peritonitis group compared with those in the SC group; these levels were significantly reversed in the LPS-induced peritonitis + dulaglutide group. In conclusion, dulaglutide may effectively maintain peritoneal integrity primarily by suppressing inflammation, oxidative stress, EMT and fibrosis.

BACKGROUND:Atherogenic dyslipidemia remains an essential predictor of cardiovascular disease. This study tested whether combined dapagliflozin and alirocumab therapy could exert a synergistic effect on reducing dyslipidemia by inhibiting CD36 expression. MATERIALS AND RESULTS:In vitro experiments revealed that a stepwise increase in glyceryl trioleate (TG) or oxidized low-density lipoprotein (ox-LDL) concentrations led to increased lipid-droplet formation (LDF) in macrophages. Silencing receptor CD36, but not the LDL receptor, as well as dapagliflozin/alirocumab therapy, significantly attenuated LDF in macrophages. Lipid droplets were transferred from TG or ox-LDL-treated macrophages to untreated macrophages using tunneling nanotubes. Combined dapagliflozin/alirocumab therapy was superior to monotherapy in upregulating lipoprotein lipase activity (LPLa) and suppressing the expression levels of receptor CD36, angiopoietin-4 (ANGPT4), LDF, proinflammatory cytokines, oxidative-stress markers, and Peroxisome proliferator-activated receptor gamma (PPARγ) in macrophages and hepatocytes. This was achieved by downregulating Tyrosine-protein kinase Fyn (Fyn)/proto-oncogene tyrosine-protein kinase Src (Src)/Mitogen-Activated Protein Kinase (MAPK)-family signaling. In vivo, db/db mice were categorized into groups 1 [ db/db + high-fat diet (HFD)]/2 ( db/db + HFD + dapagliflozin)/3 ( db/db + HFD + alirocumab)/4 ( db/db + HFD + combined dapagliflozin/alirocumab). After 12 weeks of HFD feeding, circulatory levels of angiopoietin-like protein 4, TG, total cholesterol, and LDL, as well as the number of aortic atheromas, inflammatory cell count, aortic tension, and circulatory proinflammatory cytokine levels were all significantly higher in group 1, while these were significantly reversed in groups 2/3, and further significantly reversed in group 4. Meanwhile, body weight, blood sugar, HB1AC, and LPLa exhibited an opposite pattern among the groups (all P < 0.0001). CONCLUSION:Receptor CD36 plays a critical role in LDF/atherogenic dyslipidemia, which were synergistically suppressed by combined dapagliflozin-alirocumab therapy.

Background: This study tested the hypothesis that extracorporeal shockwave therapy (ECSWT) effectively rescues critical limb ischemia (CLI) in mice through the upregulation of GPR120, which protects against inflammation and angiogenesis to restore blood flow in the ischemic area. Methods and results: Compared with the control, ECSWT-induced GPR120-mediated anti-inflammatory effects significantly suppressed the expression of inflammatory signaling biomarkers (TAK1/MAPK family/NF-κB/IL-1β/IL-6/TNF-α/MCP-1) in HUVECs, and these effects were abolished by silencing GPR120 or by the GPR120 antagonist AH7614 (all P < 0.001). C57BL/6 mice (n = 40) were equally categorized into Groups 1 (sham-operated control), 2 (CLI), 3 (CLI + ECSWT), and 4 (CLI + ECSWT + AH7614). By Days 7, 14, and 28 just prior to harvesting the quadriceps muscle, the laser Doppler results showed that the ratio of ischemia to normal blood flow (INBF) in the CLI area was highest in Group 1, lowest in Group 2, and significantly greater in Group 3 than in Group 4 (all P < 0.0001). Endothelial cell markers (CD31/vWF) and GPR120 + cells exhibited identical patterns of INBF among the groups, whereas angiogenesis biomarkers (CXCR4/SDF-1/VEGF/VEGFR2) were significantly and progressively upregulated from Groups 1 to 4 (all P < 0.0001). The protein levels of inflammation (MMP-9, IL-6, and TNF-α) and oxidative stress (NOX-1 and NOX-2) and the cellular levels of inflammation (CD68+)/DNA damage (γ-H2AX+) displayed opposite patterns, whereas the small vessel density in the CLI area displayed an identical pattern of INBF among the groups (all P < 0.0001). Conclusions: ECSWT rescued CLI by increasing GPR120-mediated suppression of inflammation and enhancing angiogenesis via activation of VEGFR2.

This study revealed that adipose-derived mesenchymal stem cell-facilitated ciprofloxacin therapy effectively protected the kidney parenchyma and functional integrity against acute pyelonephritis damage in rodents. In vitro studies revealed that adipose-derived mesenchymal stem cell-derived media significantly suppressed the number of bacterial colony formation units (P < 0.0001). Additionally, the combination of adipose-derived mesenchymal stem cell-ciprofloxacin was superior to either treatment alone on suppressing lipopolysaccharide-induced inflammatory reactions in macrophages and peripheral blood-derived mononuclear cells and attenuated lipopolysaccharide-induced apoptosis/DNA damage in uroepithelial cells (Simian virus Hydrologic Unit Code 1) (all P < 0.0001). Sprague-Dawley rats were categorized into groups 1 (sham-control)/2 (acute pyelonephritis)/3 (acute pyelonephritis-ciprofloxacin)/4 (acute pyelonephritis- adipose-derived mesenchymal stem cell)/5 (acute pyelonephritis-adipose-derived mesenchymal stem cell-ciprofloxacin), and kidneys were harvested by day 5 after acute pyelonephritis induction. The in vivo results revealed that the day-5 mortality rate and creatinine levels at days 2 and 5 were significantly greater in group 2 than in groups 1 and 5 (P = 0.01), whereas the kidney injury score and inflammatory cell infiltration in the kidney were highest in group 2, lowest in group 1, and significantly greater in groups 3 and 4 than in group 5; however, there was no difference between groups 3 and 4 (all P < 0.0001). The upstream inflammatory signaling (toll-like receptor-4, myeloid differentiation primary response 88, tumor necrosis factor receptor associated factor 6 and nuclear factor-kappa B) and downstream inflammatory signaling (tumor necrosis factor-alpha, interleukin-1 beta and interleukin-6) biomarkers exhibited identical patterns of kidney injury scores among the groups (all P < 0.0001). The results of the present study showed that adipose-derived mesenchymal stem cell-facilitated ciprofloxacin reduced inflammatory signaling-induced kidney parenchymal damage and acute pyelonephritis-induced mortality and preserved kidney function.

Background: Diabetic kidney disease (DKD) is one of the most significant public health burdens worldwide. This study explored the renal protections of combined adipose-derived mesenchymal stem cells (ADMSCs) and empagliflozin (EMPA) in DKD rats. Methods: Adult-male-SD rats were equally allocated into group 1 (sham-operated-control), group 2 (DKD), group 3 (DKD + EMPA/20 mg/kg/day since day-14 after CKD-induction), group 4 [DKD + ADMSCs (6.0 x 10(5)/ intrarenal-arterial-injection/post-day-28, followed by 1.2 x 10(6)/intravenous injection post-days 35 and 42 after CKD-induction, i.e., defined as repeated administration)] and group 5 (DKD + ADMSCs + EMPA) and kidney was harvested post-day-60 CKD-induction. Results: The result showed that the blood sugar and circulatory levels of BUN/creatinine and the ratio of urine protein/creatinine at day 60 were greatly increased in group 2 as compared the SC (i.e., group 1), significantly increased in groups 3 and 4 than in groups 5, but these parameters showed the similar manner in groups 3 and 4, except for blood sugar that was significantly lower in group 3 than in group 4 (all p < 0.0001). The protein levels of inflammation (NF-kappa B/FNF-alpha/MMP-9)/oxidative-stress (NOX-1/NOX-2/oxidized protein/p22-phox)/apoptosis (cleaved-caspase-3/cleaved-PARP/mitochondrial-Bax)/fibrosis (TGF-beta/Smad 3)/mitochondrial/DNA-damaged (p-DRP1/gamma-H2AX) biomarkers revealed a similar manner of creatinine level among the groups (all p < 0.0001). Kidney injury score/fibrotic area/oxidative-stress score (8-OHdG) and cellular levels of kidneydamaged biomarkers (KIM-1/gamma-H2AX) showed a unanimous manner. In contrast, the cellular expressions of podocyte coponents (ZO-1/synaptopodin) revealed an antithetical manner of creatinine among the groups (all p < 0.0001). Conclusion: Combined ADMSCs-EMPA was superior to just one therapy for protecting kidney function and ultra structural integrity in DKD rodents.

Background: This study tested whether combined dapagliflozin (DAPA) and roxadustat (ROX) therapy was superior to a singular therapy in protecting heart and kidney functions in rats with cardiorenal syndrome (CRS). Methods and results: An in vitro study demonstrated that the cell survival (PI3K/Akt/mTOR)/cell stress (ERK1/2, JNK/p-38) signaling was significantly activated by combination therapy with ROX-DAPA (all p<0.001). Additionally, these two signaling pathways further significantly upregulated the hypoxia-induced factor (HIF)-1α which, in turn, significantly upregulated Nrf2/ARE (HO-1/NQO-1) and angiogenesis/cell-growth factors (EPO/SDF-1α/VEGF/FGF/IGF-2) and downregulated hypoxia-inducible factor prolyl-4-hydroxylase-1 (all p<0.001). Adult-male SD rats were categorized into Groups 1 (sham-operated control)/2 (CRS)/3 (CRS+ROX)/4 (CRS+DAPA)/5 (CRS+ROX+DAPA). By Day 60 after rodent CRS induction, the levels of BUN/creatinine and the ratio of urine protein to creatinine were lowest in Group 1, highest in Group 2, and significantly lower in Group 5 than in Groups 3 and 4; however, they were similar in the latter two groups, whereas the left-ventricular-ejection-fraction exhibited the opposite trend of creatinine among the groups (all p<0.0001). The protein expression levels of cell-survival (p-PI3K/p-Akt-p-mTOR)/cell-stress (p-JNK/p-p38/p-ERK1/2)/Nrf2-ARE (HO-1/NQO-1/SIRT1/SIRT3) signaling factors and angiogenesis factors (HIF-1α/VEGF/SDF-1α/FGF/IGF-2/EPO) significantly and progressively increased from Groups 1–5 (all p<0.0001). Conclusion: Combined DAPA-ROX therapy has a synergistic effect on protecting heart and kidney functions against CRS-induced damage in rodents.

Abstract Background This phase II randomized controlled trial tested whether the intracarotid arterial administration (ICAA) of autologous CD34 + cells to patients within 14 ± 7 days after acute ischemic stroke (IS) could be safe and further improve short- and long-term outcomes. Methods Between January 2018 and March 2022, 28 consecutive patients were equally randomly allocated to the cell-treated group (CD34 + cells/3.0 × 107/patient) or the control group (receiving optimal medical therapy). CD34 + cells were transfused into the ipsilateral brain infarct zone of cell-treated patients via the ICAA in the catheterization room. Results The results demonstrated 100% safety and success rates for the procedure, and no long-term tumorigenesis was observed in cell-treated patients. In cell-treated patients, the angiogenesis capacity of circulating endothelial progenitor cells (EPCs)/Matrigel was significantly greater after treatment than before treatment with granulocyte colony-stimulating factor (all p < 0.001). Blood samples from the right internal jugular vein of the cell-treated patients presented significantly greater levels of the stromal cell-derived factor 1α/EPC at 5, 10 and 30 min compared with 0 min (all p < 0.005). The National Institute of Health Stroke Scale scores were similar upon presentation, but a greater response was observed by Days 30 and 90 in the cell-treated group than in the control group. Tc-99 m brain perfusion was significantly greater at 180 days in the cell-treated group than in the control group (p = 0.046). The combined long-term end points (defined as death/recurrent stroke/or severe disability) were notably lower in the control group compared with the cell-treated group (14.3% vs. 50.0%, p = 0.103). Conclusion Intracarotid transfusion of autologous CD34 + cells is safe and might improve long-term outcomes in patients with acute IS. Trial registration ISRCTN, ISRCTN15677760. Registered 23 April 2018- Retrospectively registered, https://doi.org/10.1186/ISRCTN15677760