Cardiovascular diseases (CVDs) are a leading cause of death but are largely preventable. Consequently, devising effective predictive models to identify high-risk factors, and subsequently shape earlier interventional strategies, are of great importance. Machine-learning algorithms have been identified as a tool to devise such models, as they have greater flexibility and scalability versus traditional ones. This study aimed to identify predictive features for CVD risk, by applying 5 machine learning algorithms to the China Health and Retirement Longitudinal Study (CHARLS) dataset. 91,232 individuals from CHARLS, collected in 2011, 2012, 2015, and 2016, were randomly divided, in a 4:1 ratio, into training (72,985; 12,433 with CVD) and testing (18,247; 3,109 with CVD) datasets. This dataset was preprocessed by imputing missing values and standardizing features using StandardScaler, which was first fitted on the training dataset, then applied to training and testing sets, resulting in 61 features incorporated into 5 machine learning algorithms: logistic regression, random forest, eXtreme gradient boosting, light gradient-boosting machine (LightGBM), and deep neural network. The optimal hyperparameters and classification thresholds were identified for each algorithm, and their predictive capabilities assessed by receiver operating characteristic (ROC) curves. The top 15 predictive features from each algorithm were identified by Shapley Additive exPlanations (SHAP), followed by Venn diagrams and UpSet plots to determine which features were shared among all 5 algorithms. Out of the 61 predictors, 9 were continuous (ex. blood pressure, biomarker levels), 6 ordinal (ex. education, self‑rated health), and 46 binary (ex. smoking status, comorbidity flags). Furthermore, LightGBM had the highest area under the curve, 0.8002 (95

PURPOSE:CACNA1C variants have been identified in cardiac arrhythmias and developmental disorders (DD). Here, we aimed to explore the association between CACNA1C and epilepsy and the mechanism underlying phenotypic heterogeneity. METHODS:Trio-based whole-exome sequencing was performed in patients with focal epilepsy. Genes with recurrently identified variants were selected for further studies. RESULTS:Three de novo CACNA1C variants were identified in three patients with focal epilepsy. All variants presented no minor allele frequency in the Genome Aggregation Database, which was significantly lower than that of benign variants in the ClinVar database. The three de novo variants were evaluated as pathogenic or likely pathogenic variants according to the updated American College of Medical Genetics and Genomics (ACMG) classification. Two patients experienced infrequent seizures and became seizure free. The other patient had refractory epilepsy with DD and arrhythmia. Previously, 138 CACNA1C variants were reported to be associated with arrhythmias, of which 39 were classified as pathogenic or likely pathogenic; 30 variants were associated with DD, 19 of which were classified as pathogenic or likely pathogenic according to the updated ACMG classification. Further analysis indicated that null variants were more common in DD than in epilepsy (p=1.39×10-3) and arrhythmias (p=6.31×10-7). The epilepsy-associated variants were predominantly located in the voltage sensor region (p=9.03×10-3), whereas the arrhythmia-associated variants were mostly located in the linker region (p=2.13×10-4). There was a greater percentage of de novo variants in patients with epilepsy and DD than in those with arrhythmias (p=6.02×10-3; p=2.47×10-2). CONCLUSION:CACNA1C is potentially a candidate causative gene of focal epilepsy. The genotype-phenotype correlation of CACNA1C helps explain phenotypic heterogeneity.

BACKGROUND:The ATP2B1 gene encodes the plasma membrane calcium-transporting ATPase 1 (PMCA1), which is predominantly expressed in the brain and plays an essential role in intracellular calcium homeostasis, potentially affecting neuronal excitability and synaptic transmission. Previous studies have identified ATP2B1 variants in patients with neurodevelopmental disorders. However, the significance of ATP2B1 variants in epilepsy remains unknown. METHODS:Trio-based whole-exome sequencing (WES) was performed in the cohorts of patients with epilepsy without acquired etiologies. To define the gene-disease association, we reviewed previously reported variants of ATP2B1 and analyzed the damaging effects of the variants, genotype-phenotype correlation, and spatial-temporal expression pattern. RESULTS:ATP2B1 variants were identified in five individuals affected by generalized epilepsy or genetic epilepsy with febrile seizures plus from two unrelated families. The identified ATP2B1 variants included one de novo heterozygous missense variant (c.2920A>G/p.Ile974Val) and one co-segregated heterozygous missense variant (c.76G>A/p.Asp26Asn). The de novo variant was absent in any public population database and was evaluated as likely pathogenic according to the criteria of the American College of Medical Genetics and Genomics (ACMG). The co-segregated variant had an extremely low minor allele frequency (MAF = 7.955×10-6) in gnomAD. Computational modelling analysis suggested that the two variants led to impairment in protein stability. All of the patients had favourable outcomes without neurodevelopmental disorders. Further analysis revealed that neurodevelopmental disorders-associated variants were either biallelic variants or monoallelic truncation/missense variants, characterized by significant alterations in hydrogen bonding and/or hydrophobicity, along with extremely high AlphaMissense scores; while epilepsy-associated variants were missense variants with relatively little damage, resulting in no changes in hydrogen bonding or hydrophobicity. Notably, missense variants located in the functional domain with severe damaging effect were more likely to manifest as neurodevelopmental disorders comorbid with epilepsy. Additionally, ATP2B1 exhibits relatively low baseline expression at early development stages but significantly increases in later stages, with particularly high expression in the striatum, which is consistent with the generalized epilepsy phenotype observed in this study, and explains that severe damage variants were associated with early-onset neurodevelopmental disorders, whereas milder damage variants were associated with epilepsy. SIGNIFICANCE:ATP2B1 is a potential candidate gene for generalized epilepsy. Genetic dependent stage (GDS), gene tissue-specific expression and genotype-phenotype correlation contribute to the explanation of phenotypic heterogeneity.

[This corrects the article DOI: 10.3389/fnmol.2022.889534.].

The multiple PDZ domain crumbs cell polarity complex component gene(MPDZ;MIM:603785),is highly expressed in the brain across the whole lifespan.It encodes the multiple PDZ domain protein,which is a member of the NMDAR signaling complex that may play a role in the control of AMPAR potentiation and synaptic plasticity in excitatory synapses.1 Previously,MPDZ variants have been demon-strated to be associated with autosomal recessive congen-ital hydrocephalus-2(HYC2;MIM:615219)which is commonly complicated by brain abnormalities and devel-opmental delay.Seizures were reported in only one case.The association between MPDZ and epilepsy requires clarification.

PurposeTo identify novel genetic causes of febrile seizures (FS) and epilepsy with febrile seizures plus (EFS+).MethodsWe performed whole-exome sequencing in a cohort of 32 families, in which at least two individuals were affected by FS or EFS+. The probands, their parents, and available family members were recruited to ascertain whether the genetic variants were co-segregation. Genes with repetitively identified variants with segregations were selected for further studies to define the gene-disease association.ResultsWe identified two heterozygous ATP6V0C mutations (c.64G > A/p.Ala22Thr and c.361_373del/p.Thr121Profs*7) in two unrelated families with six individuals affected by FS or EFS+. The missense mutation was located in the proteolipid c-ring that cooperated with a-subunit forming the hemichannel for proton transferring. It also affected the hydrogen bonds with surround residues and the protein stability, implying a damaging effect. The frameshift mutation resulted in a loss of function by yielding a premature termination of 28 residues at the C-terminus of the protein. The frequencies of ATP6V0C mutations identified in this cohort were significantly higher than that in the control populations. All the six affected individuals suffered from their first FS at the age of 7–8 months. The two probands later manifested afebrile seizures including myoclonic seizures that responded well to lamotrigine. They all displayed favorable outcomes without intellectual or developmental abnormalities, although afebrile seizures or frequent seizures occurred.ConclusionThis study suggests that ATP6V0C is potentially a candidate pathogenic gene of FS and EFS+. Screening for ATP6V0C mutations would help differentiating patients with Dravet syndrome caused by SCN1A mutations, which presented similar clinical manifestation but different responses to antiepileptic treatment.

Purpose Inflammation associated endothelial cell (EC) dysfunction is key to atherosclerotic disease. Recent studies have demonstrated a protective role of amitriptyline in cardiomyocytes induced by hypoxia/reoxygenation. However, the mechanism by which amitriptyline regulates the inflammatory reaction in ECs remains unknown. Thus, the aim of this study was to investigate whether amitriptyline protects against inflammation in TNF-α-treated ECs. Methods HUVECs were incubated with amitriptyline (2.5 μM) or TNF-α (20 ng/ml) for 24 h. EdU, tube formation, transwell, DHE fluorescence staining, and monocyte adhesion assays were performed to investigate endothelial function. Thoracic aortas were isolated from mice, and vascular tone was measured with a wire myograph system. The levels of ICAM-1, VCAM-1, MCP-1, phosphorylated MAPK and NF-κB were detected using western blotting. Results Amitriptyline increased the phosphorylation of nitric oxide synthase (eNOS) and the release of NO. Amitriptyline significantly inhibited TNF-α-induced increases in ASMase activity and the release of ceramide and downregulated TNF-α-induced expression of proinflammatory proteins, including ICAM-1, VCAM-1, and MCP-1 in ECs, as well as the secretion of sICAM-1 and sVCAM-1. TNF-α treatment obviously increased monocyte adhesion and ROS production and impaired HUVEC proliferation, migration and tube formation, while amitriptyline rescued proliferation, migration, and tube formation and decreased monocyte adhesion and ROS production. Additionally, we demonstrated that amitriptyline suppressed TNF-α-induced MAPK phosphorylation as well as the activity of NF-κB in HUVECs. The results showed that the relaxation response of aortic rings to acetylcholine in the WT-TNF-α group was much lower than that in the WT group, and the sensitivity of aortic rings to acetylcholine in the WT-TNF-α group and WT-AMI-TNF-α group was significantly higher than that in the WT-TNF-α group. Conclusion These results suggest that amitriptyline reduces endothelial inflammation, consequently improving vascular endothelial function. Thus, the identification of amitriptyline as a potential strategy to improve endothelial function is important for preventing vascular diseases.

Purpose Previously, mutations in the voltage-gated calcium channel subunit alpha1 A (CACNA1A) gene have been reported to be associated with paroxysmal disorders, typically as episodic ataxia type 2. To determine the relationship between CACNA1A and epilepsies and the role of molecular sub-regional on the phenotypic heterogeneity. Methods Trio-based whole-exome sequencing was performed in 318 cases with partial epilepsy and 150 cases with generalized epilepsy. We then reviewed all previously reported CACNA1A mutations and analyzed the genotype-phenotype correlations with molecular sub-regional implications. Results We identified 12 CACNA1A mutations in ten unrelated cases of epilepsy, including four de novo null mutations (c.2963_2964insG/p.Gly989Argfs*78, c.3089 + 1G > A, c.4755 + 1G > T, and c.6340-1G > A), four de novo missense mutations (c.203G > T/p.Arg68Leu, c.3965G > A/p.Gly1322Glu, c.5032C > T/p.Arg1678Cys, and c.5393C > T/p.Ser1798Leu), and two pairs of compound heterozygous missense mutations (c.4891A > G/p.Ile1631Val& c.5978C > T/p.Pro1993Leu and c.3233C > T/p.Ser1078Leu&c.6061G > A/p.Glu2021Lys). The eight de novo mutations were evaluated as pathogenic or likely pathogenic mutations according to the criteria of American College of Medical Genetics and Genomics (ACMG). The frequencies of the compound heterozygous CACNA1A mutations identified in this cohort were significantly higher than that in the controls of East Asian and all populations (P = 7.30 × 10–4, P = 2.53 × 10–4). All of the ten cases were ultimately seizure-free after antiepileptic treatment, although frequent epileptic seizures were observed in four cases. Further analysis revealed that episodic ataxia type 2 (EA2) had a tendency of higher frequency of null mutations than epilepsies. The missense mutations in severe epileptic phenotypes were more frequently located in the pore region than those in milder epileptic phenotypes (P = 1.67 × 10–4); de novo mutations in the epilepsy with intellectual disability (ID) had a higher percentage than those in the epilepsy without ID (P = 1.92 × 10–3). Conclusion This study suggested that CACNA1A mutations were potentially associated with pure epilepsy and the spectrum of epileptic phenotypes potentially ranged from the mild form of epilepsies such as absence epilepsy or partial epilepsy, to the severe form of developmental epileptic encephalopathy. The clinical phenotypes variability is potentially associated with the molecular sub-regional of the mutations.

Wellens综合征最早于1982年报道.约10％ ～15％的急性冠脉综合征患者表现为Wellens综合征.Wellens综合征患者主要表现为:有心绞痛症状,心绞痛缓解时,心电图有特征性T波改变(V2-3导联T波对称性深倒或正负双向,可包括V1、V4-6),无明显ST段偏移,胸前导联R波正常,无病理性Q波,心肌坏死标志物可升高.Wellens综合征患者冠脉造影常表现为前降支近段重度狭窄,少数患者可表现为前降支近段完全闭塞,远段有较好的侧支循环形成.部分患者在胸痛发作时,可表现为一过性前壁导联ST段抬高.Wellens综合征患者有进展为急性前壁心肌梗死的风险,应尽早识别并积极救治.

Purpose Berberine was reported to exert beneficial effects on cardiac hypertrophy. However, its cellular and molecular mechanisms still remained unclear. Methods Cardiac hypertrophy was induced in male Sprague-Dawley (SD) rats by transverse aorta constriction (TAC), with or without 6-week treatment of berberine. Echocardiography was performed to evaluate cardiac function. Rats were then sacrificed for histological assay, with detection for proteins and mRNA. H9c2 cells were pretreated with berberine of different concentrations (0, 1 μM, and 10 μM), followed by treatment with 2 μM norepinephrine (NE). Cells of different groups were measured for cell surface area, with mRNA detected by qRT-PCR and proteins by western blot. Results Compared with the sham group, rats of the TAC group showed significantly increased cardiac hypertrophy and fibrosis, which could be ameliorated by treatment with berberine. Western blot showed that mammalian target of rapamycin (mTOR) signaling-related protein expressions, including phospho-mTOR, phospho-4EBP1, and phospho-p70 S6K (Thr389), but not phospho-p70 S6K (Ser371), were significantly increased in the TAC group, which were inhibited by berberine treatment. H9c2 cells were treated with NE to induce hypertrophy with increased cell surface area and mRNA expressions of anp and bnp . Berberine of 10 μM, but not 1 μM, significantly ameliorated NE-induced hypertrophy and inhibited protein expressions of mTOR signaling pathway similar to those in the rat model. Conclusions Berberine can exert cardioprotective effects on both pressure-overloaded cardiac hypertrophy and failure in vivo and NE-induced hypertrophy in vitro. Our results suggest berberine could be a potential treatment for patients with cardiac hypertrophy and failure.

Estrogen deficiency induces cardiac dysfunction and increases the risk of cardiovascular disease in postmenopausal women and in those who underwent bilateral oophorectomy. Previous evidence suggests that puerarin, a phytoestrogen, exerts beneficial effects on cardiac function in patients with cardiac hypertrophy. In this study, we investigated whether puerarin could prevent cardiac hypertrophy and remodeling in ovariectomized, aortic-banded rats. Female SD rats subjected to bilateral ovariectomy (OVX) plus abdominal aortic constriction (AAC). The rats were treated with puerarin (50 mg·kg−1 ·d−1, ip) for 8 weeks. Then echocardiography was assessed, and the rats were sacrificed, their heart tissues were extracted and allocated for further experiments. We showed that puerarin administration significantly attenuated cardiac hypertrophy and remodeling in AAC-treated OVX rats, which could be attributed to activation of PPARα/PPARγ coactivator-1 (PGC-1) pathway. Puerarin administration significantly increased the expression of estrogen-related receptor α, nuclear respiratory factor 1, and mitochondrial transcription factor A in hearts. Moreover, puerarin administration regulated the expression of metabolic genes in AAC-treated OVX rats. Hypertrophic changes could be induced in neonatal rat cardiomyocytes (NRCM) in vitro by treatment with angiotensin II (Ang II, 1 μM), which was attenuated by co-treatemnt with puerarin (100 μM). We further showed that puerarin decreased Ang II-induced accumulation of non-esterified fatty acids (NEFAs) and deletion of ATP, attenuated the Ang II-induced dissipation of the mitochondrial membrane potential, and improved the mitochondrial dysfunction in NRCM. Furthermore, addition of PPARα antagonist GW6471 (10 μM) partially abolished the anti-hypertrophic effects and metabolic effects of puerarin in NRCM. In conclusion, puerarin prevents cardiac hypertrophy in AAC-treated OVX rats through activation of PPARα/PGC-1 pathway and regulation of energy metabolism remodeling. This may provide a new approach to prevent the development of heart failure in postmenopausal women.

Phenotype transition of vascular smooth muscle cells (VSMCs) is implicated in vascular diseases. Angiotensin-converting enzyme 2 (ACE2) is a perspective cardiovascular target due to its ability of converting angiotensin (Ang II) to Ang (1–7). Our group recently showed that ACE2 can regulate the function of endothelial progenitor cell–derived exosomes (EPC-EXs). Here, we investigate whether ACE2 could affect the role of EPC-EXs on phenotype transition of VSMCs. After co-incubation with EXs released from EPC overexpressed ACE2 (EPC-EXsACE2), the ACE2 level and Ang II/Ang (1–7), proliferation/migration, phenotype gene, cytokine and NF-κB level on VSMCs were assessed. To determine the EX uptake route, VSMCs were pretreated with inhibitors. We found that (1) EPC-EXs and EPC-EXsACE2 were uptaken by VSMCs dominantly through caveolin-dependent endocytosis. (2) EPC-EXsACE2 remarkably increased the ACE2 level and decreased Ang II/Ang (1–7) in VSMCs activated by Ang II, whereas EPC-EXsACE2 pretreated by proteinase A blocked this effect. (3) EPC-EXsACE2 had better effects than EPC-EXs on reducing proliferation/migration activities and cytokine (MCP-1, TNF-α) secretion of Ang II–activated VSMCs. (4) EPC-EXs attenuated Ang II–induced VSMC synthetic phenotype change as evidenced by upregulated expressions of calponin and a-SMA and downregulated expressions of CRBP-1 and MYH10, associated with a decreased NF-κB level. EPC-EXsACE2 augmented these effects, which were attenuated by ACE2 inhibitor (DX600). In conclusion, EPC-EXsACE2 reduced Ang II–induced VSMC phenotype change by conveying functional ACE2 to downregulate the activated NF-κB pathway.

Endothelial progenitor cell (EPC) transplantation has shown advantages in the treatment of myocardial infarction (MI) in animal models and clinical trials through mechanisms of direct intercellular contacts, autocrine, and paracrine. However, the effects of EPC transplantation for MI treatment remain controversial and the underlying mechanisms have not been fully elucidated. Here, we explored the role of Rab27a in the therapeutic potential of EPC transplantation in MI. We found that Rab27a knockout impaired the viability, and reduced the proliferation and tube formation function of ECPs. The recovery of cardiac function and improvement of ventricular remodeling from EPCs transplantation were significantly damaged by Rab27a deletion in vivo. Rab27a deletion inhibited the protein expression of phosphoinositide 3-kinase (PI3K) and cyclin D1 and the phosphorylation levels of Akt and FoxO3a. Therefore, Rab27a knockout suppressed the PI3K-Akt-FoxO3a/cyclin D1 signaling pathway. Furthermore, Rab27a ablation dramatically reduced exosome release in EPCs. These results demonstrated that Rab27a plays an essential role in EPC functions. The elucidation of this mechanism provides novel insights into EPC transplantation as a promising treatment for post-MI injuries.

目的 探讨毛蕊异黄酮对大鼠心肌肥厚的保护作用及潜在机制.方法 使用腹主动脉缩窄(TAAC)制备大鼠心肌肥厚模型,将SD大鼠随机分为TAAC组、溶剂对照组(vehicle组)以及毛蕊异黄酮处理组(calycosin组),术后1周每天腹腔给药.8周后行超声心动图检查,检测左心室有创压以及心脏组织中JAK1、STAT3蛋白表达量.结果 8周后,与TAAC组、vehicle组相比较,calycosin组大鼠左心室质量指数(LVMI)、IVS、LVESD、LVESV均明显降低(P<0.05),LVEDD、LVEDV明显升高(P<0.05),LVEF无明显改变(P>0.05);calycosin组大鼠心室收缩末压及±dp/dt明显升高(P<0.05);calycosin组心肌组织JAK1、STAT3蛋白表达量明显下降,差异有统计学意义(P<0.05).而TAAC组与vehicle组比较差异无统计学意义(P>0.05).结论 毛蕊异黄酮在大鼠压力负荷心肌肥厚中起保护作用,其机制可能与抑制JAK/STAT信号通路有关.

Resveratrol has been reported to inhibit vascular smooth muscle cell proliferation and neointimal hyperplasia following arterial injury; however, the underlying mechanisms remain unclear. The present study was designed to investigate the effects of resveratrol on angiotensin II (AngII)-induced proliferation of A7r5 cells and explore the molecular mechanisms responsible for the observed effects. Resveratrol inhibited cell proliferation and migration, and decreased the AngII-induced protein expression of a-smooth muscle actin (alpha-SMA), proliferating cell nuclear antigen (PCNA) and cyclin-dependent kinase 4 (CDK4). Resveratrol inhibited Angll-induced activation of intracellular Ca2+/cal modu I i n-dependent protein kinase II (CaMKII) and histone deacetylases 4 (HDAC4), as well as blocking AngII-induced cell cycle progression from the G(0)/G(1) to S-phase. In vivo, 4-weeks of resveratrol treatment decreased the neointima area and the neointima/media area ratio in rats following carotid balloon injury. Resveratrol also inhibited the protein expression of total and phosphorylated CaMKII and HDAC4 in the injured arteries. In conclusion, the present study demonstrated that resveratrol attenuated AngThinduced cell proliferation and neointimal hyperplasia by inhibiting the CaMKII-HDAC4 signaling pathway. These findings suggest that resveratrol may potentially prevent arterial restenosis.

Objective To detect protein the expression of mammalian target of rapamycin (mTOR) signaling in pressure-overloaded cardiac hypertrophy and H9c2 cardiomyocyte hypertrophy,and investigate effect of mTOR signaling on cardiac hypertrophy.Methods Sixteen SD rats were randomly grouped as the abdominal aorta coarctation (AAC) group and the sham group,with 8 rats for each group.Two weeks after AAC,echocardiography and histological analysis were performed.Norepinephrine (NE) was used to induce H9c2 cardiomyocyte hypertrophy in vitro.Relative protein expressions were detected for mTOR signaling by western blot.Results Compared with the sham group,rats in the ACC group showed a significant increase in left ventricular posterior wall end-diastolic thickness (LVPWd),interventricular septal end-diastolic thickness (IVSd) and heart weight index (HWI) (all P ＜ 0.05).Western blot showed that there was a significant increased ratio of P-mTOR and mTOR protein expression in the AAC group compared with the sham group (P ＜0.05).NE treatment significantly increased the ratio of P-mTOR and mTOR protein expression in vitro,with a significant difference compared with the control group (P ＜ 0.05).Conclusion Protein expressions in mTOR signaling can be significantly increased in pressure-overloaded cardiac hypertrophy and H9c2 cardiomyocyte hypertrophy,mTOR signaling may play a key role in the pathogenesis of cardiac hypertrophy.

Previous evidence has suggested that puerarin may attenuate cardiac hypertrophy; however, the potential mechanisms have not been determined. Moreover, the use of puerarin is limited by severe adverse events, including intravascular hemolysis. This study used a rat model of abdominal aortic constriction (AAC)-induced cardiac hypertrophy to evaluate the potential mechanisms underlying the attenuating efficacy of puerarin on cardiac hypertrophy, as well as the metabolic mechanisms of puerarin involved. We confirmed that puerarin (50 mg/kg per day) significantly attenuated cardiac hypertrophy, upregulated Nrf2, and decreased Keap1 in the myocardium. Moreover, puerarin significantly promoted Nrf2 nuclear accumulation in parallel with the upregulated downstream proteins, including heme oxygenase 1, glutathione transferase P1, and NAD(P)H:quinone oxidoreductase 1. Similar results were obtained in neonatal rat cardiomyocytes (NRCMs) treated with angiotensin II (Ang II; 1 μM) and puerarin (100 μM), whereas the silencing of Nrf2 abolished the antihypertrophic effects of puerarin. The mRNA and protein levels of UGT1A1 and UGT1A9, enzymes for puerarin metabolism, were significantly increased in the liver and heart tissues of AAC rats and Ang II-treated NRCMs. Interestingly, the silencing of Nrf2 attenuated the puerarin-induced upregulation of UGT1A1 and UGT1A9. The results of chromatin immunoprecipitation-quantitative polymerase chain reaction indicated that the binding of Nrf2 to the promoter region of Ugt1a1 or Ugt1a9 was significantly enhanced in puerarin-treated cardiomyocytes. These results suggest that Nrf2 is the key regulator of antihypertrophic effects and upregulation of the metabolic enzymes UGT1A1 and UGT1A9 of puerarin. The autoregulatory circuits between puerarin and Nrf2-induced UGT1A1/1A9 are beneficial to attenuate adverse effects and maintain the pharmacologic effects of puerarin.

Stem cell therapy can be used to repair and regenerate damaged hearts tissue; nevertheless, the low survival rate of transplanted cells limits their therapeutic efficacy. Recently, it has been proposed that exosomes regulate multiple cellular processes by mediating cell survival and communication among cells. The following study investigates whether injured cardiomyocytes-derived exosomes (cardiac exosomes) affect the survival of transplanted bone marrow mesenchymal stem cells (BMSCs) in infarcted heart. To mimic the harsh microenvironment in infarcted heart that the cardiomyocytes or transplanted BMSCs encounter in vivo, cardiomyocytes conditioned medium and cardiac exosomes collected from H2O2-treated cardiomyocytes culture medium were cultured with BMSCs under oxidative stress in vitro. Cardiomyocytes conditioned medium and cardiac exosomes significantly accelerated the injury of BMSCs induced by H2O2; increased cleaved caspase-3/caspase-3 and apoptotic percentage, and decreased the ratio of Bcl-2/Bax and cell viability in those cells. Next, we explored the role of cardiac exosomes in the survival of transplanted BMSCs in vivo by constructing a Rab27a knockout (KO) mice model by a transcription activator-like effector nuclease (TALEN) genome-editing technique; Rab27a is a family of GTPases, which has critical role in secretion of exosomes. Male mouse GFP-modified BMSCs were implanted into the viable myocardium bordering the infarction in Rab27a KO and wild-type female mice. The obtained results showed that the transplanted BMSCs survival in infarcted heart was increased in Rab27a KO mice by the higher level of Y-chromosome Sry DNA, GFP mRNA, and the GFP fluorescence signal intensity. To sum up, these findings revealed that the injured cardiomyocytes-derived exosomes accelerate transplanted BMSCs injury in infarcted heart, thus highlighting a new mechanism underlying the survival of transplanted cells after myocardial infarction.

目的:探讨抑制骨髓间充质干细胞外泌体的释放对其生物学特性的影响及其机制.方法:利用TALEN技术靶向性敲除Rab27a基因构建外泌体释放缺失小鼠模型,随后分离、培养并鉴定骨髓间充质干细胞,用外泌体提取试剂盒提取培养基中的外泌体,并用纳米颗粒跟踪分析(NTA)计数外泌体的数量,透射电镜观察外泌体的大小及形态.利用EdU实验及检测增殖细胞核抗原(PCNA)的表达来评价间充质干细胞的增殖能力.通过TUNEL染色及MTS实验检测间充质干细胞对缺氧的耐受能力.结果:敲除Rab27a的间充质干细胞释放外泌体的数量明显减少,抑制间充质干细胞外泌体的释放能显著降低其增殖及对缺氧的耐受能力.结论:抑制间充质干细胞外泌体的释放可导致其增殖能力及对缺氧的耐受能力明显降低.