Babao Dan (BBD), a traditional Chinese herbal compound, has demonstrated significant antitumor effects and is clinically used as an adjunctive therapy for various gastrointestinal malignancies. However, its underlying mechanisms of action remain poorly understood. This study aimed to investigate the effects of BBD on 5-fluorouracil (5-FU)-induced myelosuppression and T-lymphocyte subpopulation alterations, and to explore its mechanism in ameliorating cellular immune dysfunction caused by myelosuppression via the MAPK signaling pathway. The major constituents of BBD are fatty acids, triterpenoids, flavonoids, and alkaloids, along with other compounds. BBD significantly improved survival, reduced body weight loss, and mitigated the decline in splenic index in mice following 5-FU-induced chemotherapy. Additionally, it alleviated the reduction of peripheral blood cells, including leukocytes, neutrophils, and reticulocytes, and attenuated 5-FU-induced bone marrow hematopoiesis suppression. Furthermore, BBD restored the imbalance in the proportions of Th1 and Th2 cells within the spleens of 5-FU-induced mice. It also reversed the inhibition of CD25+ and CD69+ expression, the reduction in lymphocyte division generation, and the decreased expression of Ki67 and PCNA, thereby enhancing T-cell activation and proliferation in splenic cells. Moreover, BBD alleviated the G1 phase blockage in the bone marrow cell cycle and upregulated the expression of key proteins, including CDK2, CDK4, CDK6, Cyclin D1, Cyclin E1, p-Rb, p-c-Fos, p-c-Jun, p-ERK, p-NF-κB p65, and p-p38, in bone marrow cells. BBD may promote the expression of G1/S checkpoint-related proteins in myeloid cells by regulating the MAPK signaling pathway, thereby upregulating the phosphorylation level of Rb and facilitating the transition from the G1 phase to the S phase. This orderly cell cycle progression enhances cell proliferation and improves the hematopoietic function of myeloid cells after 5-FU chemotherapy. Consequently, BBD restores the ratio of peripheral T-cell subsets, corrects the imbalance of Th-cell subsets, and enhances the activation and proliferation functions of CD4+T cells and CD8+T cells, ultimately rectifying the dysregulated immune homeostasis of the organism.

Following the publication of the above article, and a Corrigendum (doi: 10.3892/mmr.2025.13581) that was published to rectify errors associated with the published versions of Figs. 5, 8, 9 and 10, an interested reader contacted the Editorial Office to explain that they had identified additional issues in need of further investigation. Subsequently, the authors recognized that there were further problems associated with the manuscript requiring attention which arose while they were organizing materials related to this manuscript. To ensure the scientific integrity of the work, the authors have requested that this article now be retracted from the Journal. In view of these additional issues, the Editor of Molecular Medicine Reports has agreed to the authors' request to retract this paper. Note that all the authors agree to this retraction, and the Editor and the authors apologize to the readership of the Journal for any inconvenience caused. [Molecular Medicine Reports 29: 102, 2024; DOI: 10.3892/mmr.2024.13226].

This study investigates the mechanism by which Qiling Fuzheng Qingjie Granule (QFQ) regulates immune function following doxorubicin (DOX) chemotherapy. Network pharmacology analysis was employed to explore the relationship between QFQ and the immune system. A mouse model of DOX-induced immune dysfunction was established and treated with QFQ. Techniques including histological staining, peripheral complete blood count analysis, flow cytometry, and seahorse metabolic analysis were used to systematically evaluate the effects of QFQ on immune organs, bone marrow hematopoiesis, T-cell subset distribution and function, mitochondrial metabolism, and antitumor efficacy. The results demonstrated that QFQ significantly alleviated DOX-induced atrophy of the thymus and spleen, preserved bone marrow hematopoietic function, and reduced peripheral blood cell loss. Mechanistically, QFQ improved mitochondrial membrane potential and oxidative phosphorylation, thereby restoring T-cell metabolic capacity, enhancing T-cell activation and proliferation, and restoring cytotoxic T lymphocyte (CTL) function. Consequently, QFQ rebalanced the dynamics among Th1/Th2/Th17/Treg, Effector Memory T/Effector T cells (Tem/Teff), and Central Memory T cells (Tcm) subsets, ameliorated DOX-induced immune dysregulation. In addition, QFQ intervention not only enhanced antitumor efficacy in the A20 lymphoma mouse model but also significantly alleviated the exhaustion phenotype in CD8+T cells. In summary, this investigation establishes that QFQ enhances T-cell functional performance primarily through the regulation of mitochondrial energy metabolism. By restoring metabolic homeostasis in T lymphocytes, QFQ effectively counteracts DOX-induced immune dysfunction, highlighting its potential as a promising adjunctive therapy in chemotherapy-induced immunosuppression.

Ethnopharmacological Relevance Pien Tze Huang (PZH) is a traditional medicinal formula consisted of four traditional Chinese medicines (TCMs) including Panax notoginseng (Burk.) F. H. Chen, Snake Gall, Calculus Bovis and Moschus, with clinical efficacy against Colorectal Cancer (CRC). However, the molecular and functional mechanisms underlying this efficacy are not fully elucidated. Aims of the Study This study aimed to assess the impact of PZH on CRC cancer stem cells (CSCs), and evaluate the coordination effect of PZH on T cell-mediated anti-CRC with patient-derived autologous T cell co-culture. Materials and methods High-performance liquid chromatography (HPLC) was used to identify the main components of PZH. CCK8 and spheroid formation assays were conducted for assessing cell viability and stemness function. Western blot, immunofluorescence and immunohistochemistry were used to evaluate CSC markers and PD-L1 expression. T cell successful expansion was validated by flow cytometry. Co-culture assay was conducted to explore the activation effect of PZH on T cells. The potential mechanism of PZH in CRC was identified with transcriptomics sequencing and network pharmacology analysis. Results PZH reduced cell viability and spheroid formation ability in CRC, and suppressed the expression of CSC markers - LGR5, DCLK1, and CD133. Moreover, PZH enhanced T cell-mediated cytotoxicity against CRC cells by decreasing the expression of PD-L1. Furthermore, PZH with anti-PD-1 immunotherapy enhancing antitumor efficacy and increasing CD8+ T cell infiltration with decreasing expression of CSC markers and PD-L1. Notably, PZH inhibited CRC patient-derived organoids (PDOs) tumorigenesis and increased autologous T cell cytotoxicity against PDOs (n=5). Consistently, PZH decreased expression of CSC markers and PD-L1 in PDOs. RNA sequencing and network pharmacology also highlighted that PZH inhibited CRC stemness and PD-L1 to enhance T cell-mediated antitumor effects. Conclusions PZH enhances T cell-mediated killing by inhibiting the expression of CRC stem cell markers and PD-L1, which warrant further investigation and clinical applications.

ETHNOPHARMACOLOGICAL RELEVANCE:Qingjie Fuzheng Granules (QFG), a herbal formula, has been employed as an adjuvant therapy for colitis-associated colorectal cancer (CAC), yet the underlying mechanisms by which QFG operates remain unclear. AIMS OF THE STUDY:The aim of this study is to investigate whether the potential mechanism of QFG against CAC is associated with macrophage polarization. MATERIALS AND METHODS:Non-targeted metabolomics and molecular docking assessed potential compounds of QFG to interact with targets associated with macrophage polarization. A model of AOM/DSS-induced CAC mice was established to analyze the effects of QFG on macrophage polarization using flow cytometry and immunohistochemical staining. In vitro experiments involved models of Ana-1 macrophages, either induced by varying QFG concentrations or with MD2 knockdown, to analyze M1-like phenotype. Meanwhile, M2-like macrophages models induced by IL-4 or culture supernatant of CT26 cells were utilized to assess the effects of QFG on M2-like macrophages. Finally, the mRNA expression of M1-like phenotype related to TLR4 pathways and the protein expression in IL-4R-mediated pathways were analyzed using RT-qPCR and Western blot, respectively. RESULTS:Molecular docking confirmed the presence of binding sites between the ingredients of QFG and IL-4R or TLR4/MD2 receptor complex. QFG could induce a shift in macrophages towards an M1-like phenotype while inhibiting an M2-like phenotype in the colon with CAC mice and Ana-1 macrophages. QFG resulted in the upregulation of iNOS, IL-6, IL-1β, and TNF-α mRNA expression, which could be counteracted by TAK242, SR11302, INH14, PDTC, and LY294002, or by the knockdown of MD2. Meanwhile, QFG inhibited IL-4R-induced phosphorylation of STAT 6 and Akt. CONCLUSION:Various monomer components within QFG can bind to MD2 or IL-4R, respectively, thereby inducing macrophages towards an M1-like phenotype through TLR4-mediated NF-κB, MAPK, and PI3K/Akt pathway activation, or inhibiting macrophages towards an M2-like phenotype via IL-4R-mediated JAKs pathway inhibition, ultimately exerting an inhibitory effect on the occurrence and development of CAC.

Objective To observe the effects of electroacupuncture at Pishu and Shenshu acupoints on gut microbiota,Th17/Treg cells,and bone metabolism in bilateral ovariectomized rats,and to explore the intrinsic relationship between electroacupuncture in regulating bone metabolism and the gut microbiota-Th17/Treg axis in postmenopausal osteoporosis(PMOP).Methods A total of twenty SPF-grade 3-month-old female Sprague-Dawley rats were selected.They were subjected to bilateral ovariectomy(OVX)to establish PMOP rat models.The rats were randomly divided into two groups using a random number table method:model group and electroacupuncture group,with 10 rats in each group.The electroacupuncture group began electroacupuncture treatment 3 days after modeling,while the model group underwent daily handling similar to the electroacupuncture group without any additional inter-ventions.After 12 weeks of intervention,on the following morning,each rat had approximately 1 g of fecal sample collected into ster-ile EP tubes and stored at-80℃refrigerator.The rats were then anesthetized with 2%pentobarbital sodium(0.2 mL/100 g)via intra-peritoneal injection,and euthanized for tissue collection to conduct subsequent experiments.ELISA was used to detect the level of E2.The gut microbiota of fecal samples from both groups was analyzed using 16S rDNA technology,followed by microbial profil-ing analysis,inter-group difference analysis,and LEfSe multi-Level species difference discriminant analysis.Flow cytometry was used to determine the levels of Th17 and Treg cells in the colonic mesenteric lymph nodes.ELISA was also used to measure bone metabolism-related biochemical markers,including procollagen type Ⅰ N-terminal propeptide(PⅠNP),β-collagen degradation product(β-CTx),interleukin-17(IL-17),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),hydroxyvitamin D3(25-OH-D3),interleukin-10(IL-10),and transforming growth factor-β(TGF-β).Bone mineral density was measured using dual-energy X-ray absorptiometry(DXA).Results Compared with the model group,the estradiol(E2)content in peripheral blood increased in the electroacupuncture group(P<0.05).Compared with the model group,the gut microbiota health index(GMHI)significantly increased in the electroacupuncture group(P<0.05);compared with the model group,the microbiota dysbiosis index(MDI)value significantly decreased in the electroacupuncture group(P<0.05);compared with the model group,the abundance of Lactobacillaceae and Bifido-bacteriaceae in the gut microbiota increased in the electroacupuncture group(P<0.05);compared with the model group,beneficial bacteria such as Bifidobacterium and Lactobacillus were significantly enriched in the gut of the electroacupuncture group(P<0.05).Compared with the model group,the content of Th17 cells in the colonic lymph nodes was significantly reduced in the electroacu-puncture group(P<0.05);compared with the model group,the content of Treg cells in the colonic lymph nodes significantly in-creased in the electroacupuncture group(P<0.05).Compared with the model group,the expression levels of PⅠNP,β-CTx,IL-17,IL-6,and TNF-α all decreased in the electroacupuncture group(P<0.05);the expression levels of 25(OH)D3,IL-10,and TGF-β all increased(P<0.05).Compared with the model group,bone mineral density significantly increased in the electroacupuncture group(P<0.05).Conclusion Electroacupuncture at Pishu and Shenshu can increase E2 levels,upregulate the abundance of Bifidobacteria-ceae and Lactobacillaceae,and regulate the gut microbiota-Th17/Treg axis.These effects can effectively improve bone metabolic imbalance caused by estrogen deficiency,and then prevent and treat PMOP.

Knee osteoarthritis (KOA) is a chronic degenerative disease that affects the quality of life of middle‑aged and elderly individuals, and is one of the major factors leading to disability. Rongjin Niantong Fang (RJNTF) can alleviate the clinical symptoms of patients with KOA, but the molecular mechanism underlying its beneficial effects on KOA remains unknown. Using pharmacological analysis and in vitro experiments, the active components of RJNTF were analyzed to explore their potential therapeutic targets and mechanisms in KOA. The potential targets and core signaling pathways by which RJNTF exerts its effects on KOA were obtained from databases such as Gene Expression Omnibus, Traditional Chinese Medicine Systems Pharmacology and Analysis Platform. Subsequently, chondrocyte apoptosis was modeled using hydrogen peroxide (H2O2). Cell Counting Kit‑8 assay involving a poly [ADP‑ribose] polymerase‑1 (PARP1) inhibitor, DAPI staining, reverse transcription‑quantitative PCR, Annexin V‑FITC/PI staining and flow cytometry, western blotting and co‑immunoprecipitation analysis were used to determine the therapeutic efficacy of RJNTF on KOA and to uncover the molecular mechanism. It was found that PARP1‑knockdown lentivirus, incubation with PARP1 inhibitor PJ34, medium and high doses of RJNTF significantly reduced H2O2‑induced chondrocyte apoptosis. Medium and high doses of RJNTF downregulated the expression of cleaved caspase‑3, cleaved PARP1 and PAR total proteins, as well as nucleus proteins of apoptosis‑inducing factor (AIF) and migration inhibitory factor (MIF), and upregulated the expression of caspase‑3, PARP1 total protein, as well as the cytoplasmic expression of AIF and MIF, suggesting that RJNTF may inhibit chondrocyte apoptosis through the PARP1/AIF signaling pathway.

Objective: Postmenopausal osteoporosis (PMOP) is a common bone metabolic disorder in middle-aged and elderly women, yet its pathogenesis remains unclear. This study investigates the effect of nuclear factor erythroid 2-related factor 2 (Nrf2) deficiency on bone homeostasis to provide insight into the mechanisms underlying PMOP. Methods: Sixteen female SD rats were randomly assigned to Sham and ovariectomized (OVX) groups. After 12 weeks, bone homeostasis disruption and Nrf2-mediated oxidative stress responses in bone tissue cells were assessed. Nrf2 expression was modulated in UMR-106 osteoblast-like cells and RAW264.7 macrophage-derived osteoclast precursor cells through knockdown or pharmacological activation. The effects on osteogenic function and osteoclast differentiation under oxidative stress were then evaluated. Results: The OVX group of rats exhibited a disruption in bone homeostasis, potentially attributable to the reduced expression of Nrf2 and its downstream antioxidant enzymes, coupled with elevated levels of oxidative stress. Nrf2 knockdown impaired osteogenic capacity in UMR-106 cells and enhanced osteoclast differentiation in RAW264.7 cells. In contrast, activation of Nrf2 using tert-butylhydroquinone (TBHQ) promoted bone formation and suppressed osteoclast differentiation and bone resorption. Conclusion: Nrf2 deficiency may contribute to PMOP by disrupting bone homeostasis. Activation of Nrf2 may represent a potential therapeutic strategy for restoring bone balance and treating PMOP.

This monthly, peer-reviewed journal includes studies devoted to molecular medicine, underscoring aspects including pharmacology, pathology, genetics, neurosciences, infectious diseases, molecular cardiology and molecular surgery. In vitro and in vivo studies of experimental model systems pertaining to the mechanisms of a variety of diseases offer researchers the necessary tools and knowledge with which to aid the diagnosis and treatment of human diseases.

Background: Gukang Capsule has been used as a complementary and alternative medicine (CAM) for the treatment of primary osteoporosis (POP) in China. The primary aim of this study was to assess the clinical effectiveness and safety of Gukang Capsule in POP patients.Methods: A systematic search was conducted across multiple academic databases including PubMed, Web of science, Cochrane Library, China National Knowledge Infrastructure, Chongqing VIP Information, and Wanfang database to identify randomized controlled trials investigating the Gukang Capsule in the treatment of POP. The screening process, data extraction, and assessment of methodological quality were conducted independently by two reviewers. Statistical analysis was performed using the Rev Man 5.3 software. Subgroup analysis was carried out through the combination of OPF. Subgroup analysis was performed according to whether OPF were combined. Stata 12.0 was used for sensitivity and bias analysis.Results: Nineteen studies were assessed that included 1804 participants. It was found that compared with the control group, the total effective rate (RR = 1.26, 95% CI, 1.20, 1.33), the Medical Outcomes Study Short-form 36 [RR = 1.26, 95% CI(1.20, 1.33)], the bone mineral density (BMD) of lumbar vertebra (SMD = 0.77, 95% CI, 0.48, 1.07), the BMD of femoral neck [SMD = 0.84, 95% CI(0.53, 1.14)], and the BMD of Ward’s triangle (SMD = 0.64, 95% CI, 0.44, 0.85) of the Gukang Capsule experimental group were higher. Compared with the control group, the fracture healing time (SMD = −2.14, 95% CI, −2.45, −1.84), the bone specific alkaline phosphatase (BALP) levels in serum (SMD = −2.00, 95% CI, −2.83, −1.17), the tartrate resistant acid phosphatase 5b (TRACP-5b) levels in serum (SMD = −2.58, 95% CI, −3.87, −1.29) of the Gukang Capsule experimental group were lower. The bone glaprotein (BGP) levels in serum (SMD = −0.22, 95% CI, −1.86, 1.43) and the adverse events (RR = 0.80, 95% CI, 0.40, 1.63) of the experimental group and the control group have no difference.Conclusion: Gukang Capsule, as a CAM for the management of POP, exhibits the potential to enhance BMD and quality of life, expedite the healing time of OPF, diminish levels of BALP and TRACP-5b, and improve the total effective rate without increasing the adverse events.Systematic Review Registration:https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42023477774, PROSPERO CRD42023477774.

ObjectiveTo investigate the therapeutic effect and mechanism of Qingjie Fuzheng Granules on 5-FU chemotherapy-induced intestinal mucositis in a mouse model of Colorectal Cancer based on Cathepsin B/NLRP3 pathway.MethodsTwenty-four male BALB/c mice were inoculated with CT26 cells subcutaneously in the axillary of the right forelimb to construct a mouse model of colorectal cancer subcutaneous xenograft. When the tumor size reached 100-300 mm3, the mice were randomly divided into the control group, 5-FU group and 5-FU+ Qingjie Fuzheng Granules group (5-FU+QFG), 8 mice in each group. The mice in the control group were given normal saline by gavage and intraperitoneal injection, while those in each other group were given intraperitoneal injection of 5-FU (50 mg/kg d-1) once daily for five successive days. The 5-FU+QFG group was given Qingjie Fuzheng Granules (1.0 g/kg d-1) by gavage at the same time of chemotherapy, once a day, for seven successive days. The diarrhea of the mice was observed every day for seven days, and the body weight and tumor volume were measured and recorded. After the last administration of the drug on the seventh day, the mice were fasted for 12 h, then, animal samples of mice were collected for experimental detection. The pathological morphology of jejunum tissue was observed by HE staining. The expressions of diamine oxidase (DAO), D-lactic acid (D-LA), endotoxin (ET), tumor necrosis factor-α(TNF-α), interleukin -1β(IL-1β) and interleukin -6(IL-6) in serum were detected by ELISA. The positive expression of Cathepsin B in the jejunum tissue was detected by immunohistochemistry. The mRNA expression levels of ZO-1 and Occludin in the jejunum tissue were detected by real-time quantitative PCR, and the protein expressions of ZO-1, Occludin, Cathepsin B, NLRP3, ASC and cleaved Caspase-1 in the jejunum tissue were detected by Western blot.ResultsDuring the experimental period, Qingjie Fuzheng Granules significantly alleviated the diarrhea symptoms induced by 5-FU (P<0.05), but did not significantly improve the body weight loss induced by 5-FU (P>0.05). There was no significant synergistic effect of 5-FU on the inhibition of tumor growth (P>0.05). QFG could significantly improve the symptoms of intestinal mucosal injury caused by 5-FU, such as shortened villi, deeper crypts, and infiltration of inflammatory cells. Compared with the 5-FU group, QFG significantly reduced the expressions of DAO, D-LA and ET as well as pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 in mouse serum (P<0.05); The protein expression of Cathepsin B in jejunum tissue of 5-FU+QFG group was significantly reduced(P<0.05), The protein expression levels of NLRP3, ASC, cleaved Caspase-1 in the jejunum tissue of 5-FU+QFG group were decreased (P<0.05). The mRNA and protein expression levels of tight junction proteins ZO-1 and Occludin in jejunum tissue of 5-FU+QFG group were increased (P<0.05);ConclusionQingjie Fuzheng Granules can reduce the release of pro-inflammatory factors, inhibit the intestinal mucosal inflammatory response and repair the intestinal mucosal barrier damage by regulating the Cathepsin B/NLRP3 pathway, thereby alleviating the chemotherapy-induced intestinal mucositis induced by 5-FU.

Post-translational modification pathway of protein ubiquitination is intricately associated with tumorigenesis. We previously reported elevated ubiquitin-conjugating enzyme 2T (UBE2T) as an independent risk factor in stage I lung adenocarcinoma and promoting cellular proliferation. However, its underlying mechanisms needed further investigation. Immunohistochemistry was used to assess the expression of UBE2T and retinoic acid receptor-related orphan receptor α (RORA) in stage I LUAD. Cell proliferation, migration, and invasion of LUAD cell lines were measured by Cell Counting Kit-8 assay (CCK-8), Colony-forming assay and Transwell assay, respectively. Western blot analysis was performed to determine the expression of epithelial-mesenchymal transition (EMT) markers. A xenograft model was established to evaluate the proliferative capacity of UBE2T and its interaction with RORA in promoting LUAD. Mechanistic insights into the promotion of early-stage LUAD by UBE2T were obtained through luciferase reporter assay, chromatin immunoprecipitation and co-immunoprecipitation. UBE2T and RORA expression was significantly up- and down-regulated in early-stage LUAD patients which’s proved to be associated with unfavorable outcomes, strengthened cell proliferation, migration, EMT and invasion through its interaction with RORA both in vivo and in vitro. The growth NSCLC xenografts was reduced by down-expression of UBE2T but was suppressed by RORA knockout. Mechanistically, UBE2T mediated the ubiquitination of the intermediate transcription factor PBX1, which played a transcriptional role in downstream regulation of RORA. The oncogenic role of UBE2T and the UBE2T-PBX1-RORA axis in driving malignant progression in Stage I LUAD had been established. UBE2T might be a novel and promising therapeutic target for LUAD treatment.

To investigate the effect of Jiedu Xiaozheng Yin (JXY) on the polarization of macrophages in colitis-associated colon cancer (CAC). An orthotopic model of CAC was established to monitor changes in the pathological state of mice. Colon length, number of colon tumors were recorded, and indices for liver, spleen, and thymus were calculated. Hematoxylin and eosin (H&E) staining was employed to observe intestinal mucosal injury and tumor formation. Immunohistochemistry (IHC) staining was utilized to investigate the effect of JXY on M1 and M2 polarization of macrophages in the colonic mucosa of CAC mice. For in vitro experiments, RT-qPCR (Reverse Transcription-quantitative PCR) and flow cytometry were used to observe the effect of JXY on various M1-related molecules such as IL-1β, TNF-α, iNOS, CD80, CD86, and its phagocytic function as well as M2-related molecules including Arg-1, CD206, and IL-10. Subsequently, after antagonizing the TLR4 pathway with antagonists (TAK242, PDTC, KG501, SR11302, LY294002), the expression of IL-6, TNF-α, iNOS, and IL-1β mRNA were detected by RT-qPCR. In vivo experiments, the results showed that JXY improved the pathological condition of mice in general. And JXY treatment decreased the shortening of colon length and number of tumors as compared to non-treated CAC mice. Additionally, JXY treatment improved the lesions in the colonic tissue and induced a polarization of intestinal mucosal macrophages towards the M1 phenotype, while inhibiting polarization towards the M2 phenotype. In vitro experiments further confirmed that JXY treatment promoted the activation of macrophages towards the M1 phenotype, leading to increased expression of IL-1β, TNF-α, iNOS, CD80, CD86, as well as enhanced phagocytic function. JXY treatment concomitantly inhibited the expression of M2-phenotype related molecules Arginase-1 (Arg-1), CD206, and IL-10. Furthermore, JXY inhibited M1-related molecules such as IL-6, TNF-α, iNOS, and IL-1β after antagonizing the TLR4 pathway. Obviously, JXY could exhibit inhibitory effects on the development of colon tumors in mice with CAC by promoting M1 polarization through TLR4-mediated signaling and impeding M2 polarization of macrophages.

Melatonin (MLT) has been shown to induce polarization of macrophages towards M2-like phenotype and inhibit polarization of macrophages towards M1-like phenotype through exogenous administration, which affects the development of many macrophage polarization-related diseases, such as infectious diseases, cardiovascular diseases, bone diseases, and tumors. However, whether endogenous melatonin has similar influences on macrophage polarization as exogenous melatonin is still under investigation. This study revealed that the process of lipopolysaccharide (LPS) inducing macrophages to polarize towards M1-like phenotype was accompanied by an increase in endogenous MLT secretion. To explore the role of increased endogenous MLT in the polarization process of macrophages, whether similar to the function of exogenous MLT in inhibiting polarization of macrophages towards M1-like phenotype, we established LPS-induced MLT deficiency models in vitro to investigate the effects of endogenous MLT on the secretion of cytokines, co-stimulatory molecules, ROS, and phagocytic function in LPS-induced M1-like macrophages. Additionally, we aimed to elucidate the mechanism by which LPS affects the secretion of endogenous MLT by macrophages. Our results confirm that LPS induces transcription of Aanat through the TLR4/TRIF pathway, consequently facilitating the secretion of MLT by macrophages. In this way, IRF3 is the main transcription factor that regulates Aanat transcription. Endogenous MLT plays a role in inhibiting the polarization of macrophages towards M1 phenotype and delaying cell apoptosis during LPS-induced polarization towards M1 phenotype. This phenomenon may be a form of self-protection that occurs when macrophages engulf pathogens while avoiding oxidative stress and apoptosis caused by LPS. This conclusion clarifies the role of endogenous MLT in the clearance of pathogens by macrophages, providing a theoretical basis for understanding its role in innate immunity.

Objective To investigate the anti-inflammatory properties of Qingjie Fuzheng granules (QFG) in vivo using a dextran sulfate sodium (DSS)–induced ulcerative colitis (UC) model and elucidate the mechanism of which QFG alleviates UC by examining T cell 17 (Th17)/regulatory T cell (Treg) balance. Methods The DSS-induced UC murine model was established, and the mice were administered QFG or saline by gavage. Their general growth characteristics, including body weight, fecal occult blood, and disease activity index were observed, and the length of the colon was recorded. Hematoxylin and eosin staining was performed to examine pathological injury within the colon tissue. The expression levels of Th17-related cytokines, Treg-related cytokines, interferon-γ (IFN-γ), indoleamine 2,3-dioxygenase 1 (IDO1), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α) in the serum were detected by enzyme-linked immunosorbent assay or Bio-Plex immunoassay. Relative mRNA expressions in the spleen and colon tissues were detected by reverse transcription–quantitative polymerase chain reaction. The protein expressions of retinoic acid–associated orphan receptor γt (RORγt), Forked head/wing helix transcription factor 3 (Foxp3), or IDO1 were detected in the spleen and colon by western blotting or immunohistochemistry. Results QFG demonstrated the potential to improve the overall pathological conditions of DSS-induced UC mice as evidenced by significantly alleviating the colon shortening and improving colon tissue pathology. QFG also decreased expressions of the pro-inflammatory cytokines IL-1β, TNF-α, IFN-γ, and interleukin-6 as well as IDO1. QFG treatment significantly reduced the expressions of Th17-related cytokines and concurrently increased the expressions of Treg-related cytokines. After QFG treatment, the expression of transcription factor RORγt decreased in the colon and spleen, while that of the transcription factor Foxp3 increased. Conclusion QFG can suppress inflammation in mice with DSS-induced UC. This effect is achieved through the regulation of transcription factors RORγt and Foxp3, which inhibits Th17 ​cell differentiation, promotes Treg cell differentiation, and maintains Th17/Treg balance.

Melatonin (MLT) is a hormone with potential anti-tumor properties, but the molecular mechanisms remain unclear. The present study aimed to explore the effect of MLT on exosomes derived from gastric cancer cells, with the goal of gaining insight into its anti-tumor activity. Results from in vitro experiments showed that MLT was able to enhance the anti-tumor activity of macrophages that had been suppressed by exosomes from gastric cancer cells. This effect was achieved through regulation of the levels of PD-L1 in macrophages via modulation of the associated microRNAs in the cancer-derived exosomes. Furthermore, MLT treatment increased the secretion of TNF-α and CXCL10 by the macrophages. Besides, MLT treatment of gastric cancer cells led to the production of exosomes that promoted the recruitment of CD8+ T cells to the tumor site, resulting in inhibition of tumor growth. Collectively, these results provide evidence for the modulation of the tumor immune microenvironment by MLT through regulation of exosomes derived from gastric cancer cells, suggesting a potential role for MLT in novel anti-tumor immunotherapies.

目的 从肝癌侧群(side population,SP)细胞的角度探讨解毒消癥饮(Jiedu Xiaozheng Yin,JXY)乙酸乙酯提取物(ethyl acetate extract from JXY,EE-JXY)抑制肝癌细胞生长的作用机制.方法 研究时间2023年1月至5月.体外培养人肝癌细胞株Huh7和PLC/PRF/5,流式细胞仪分析肝癌细胞中SP细胞的含量并分选SP细胞;平板克隆实验检测SP细胞的克隆形成能力;克隆球形成实验检测SP细胞的体外成球能力;免疫细胞荧光染色技术检测EE-JXY对肝癌SP细胞β-catenin蛋白转位的影响.结果 EE-JXY组SP细胞的含量显著低于空白组;SP细胞克隆形成能力和体外成球能力均受到显著抑制;EE-JXY使部分SP细胞β-catenin的表达从胞浆和细胞核转位到了细胞膜.结论 EE-JXY能显著抑制Huh7和PLC/PRF/5肝癌SP细胞的增殖,其机制可能与其抑制β-catenin的转位有关.

目的 探讨芪灵扶正清解方(QFQ)对Huh-7细胞能量代谢及糖酵解相关蛋白的影响.方法CCK8法检测0、62.5、125、250、500 μg/mL QFQ醇提物干预Huh-7细胞24、48和72 h后的细胞活力.将Huh-7细胞分为0、62.5、125、250、500 μg/mL组,分别予相应浓度的QFQ醇提物干预24 h,应用细胞能量代谢分析仪检测线粒体和糖酵解ATP产生速率、实时耗氧率、胞外酸化率、糖酵解速率和质子流出速率;Western blot检测Huh-7细胞缺氧诱导因子1α(HIF-1α)、己糖激酶(HK)、葡萄糖转运蛋白(Glut)1以及Glut3的蛋白表达量.结果① 与0 μg/mL组比较,125、250、500 μg/mL QFQ醇提物干预Huh-7细胞24、48和72 h后细胞活力明显降低(P＜0.01).② 与0 μg/mL组比较,62.5、125、250 μg/mL组Huh-7细胞线粒体和糖酵解ATP产生速率比值明显提高(P＜0.05),125、250 μg/mL组Huh-7细胞糖酵解速率明显降低(P＜0.05或P＜0.01);与0 μg/mL组比较,随着检测时间延长,62.5 μg/mL组Huh-7细胞实时耗氧率呈升高趋势(P＜0.05),250 μg/mL组Huh-7细胞胞外酸化率和质子流出速率呈降低趋势(P＜0.05).③ 与0 μg/mL组比较,62.5、125、250、500 μg/mL组Huh-7细胞的HIF-1α、HK、Glut1、Glut3蛋白表达量均明显降低(P＜0.01或P＜0.05).结论 QFQ通过调控细胞能量代谢和糖酵解相关蛋白HIF-1α、HK、Glut1和Glut3的表达,提高线粒体能力,抑制糖酵解能力,促进有氧呼吸,从而有效抑制Huh-7细胞的增殖.

目的 观察蒲葵子乙醇提取物(EELC)对肝癌细胞以及裸鼠皮下移植瘤的抑制作用,探讨EELC阻滞细胞周期的抗肝癌作用机制.方法 培养人肝癌HepG2细胞,采用MTT法和平板克隆形成实验检测不同浓度EELC(0、0.125、0.25、0.5 mg/mL)对HepG2细胞活力和细胞周期的影响.构建裸鼠HepG2皮下移植瘤模型,将裸鼠随机分为对照组和EELC组,分别给予生理盐水和EELC灌胃液[3 g/(kg·d)]灌胃给药,连续21 d后,剥取肿瘤组织,采用免疫组织化学方法分析肿瘤组织中PCNA、CDK1、CyclinB1和p21的蛋白表达.结果 细胞实验结果显示,与0 mg/mL组比较,0.125、0.25、0.5 mg/mL组HepG2细胞活力和克隆形成率均明显下降(P<0.05),呈剂量依赖;与0 mg/mL组比较,0.125、0.25、0.5 mg/mL组细胞周期进程明显抑制,阻滞于G2/M期.动物实验结果显示,与对照组比较,EELC组小鼠肝癌组织PCNA、CDK1和CyclinB1蛋白表达明显降低(P<0.05),p21表达明显提高(P<0.05).结论 EELC通过促进p21表达,抑制PCNA、CDK1和CyclinB1表达,使细胞周期阻滞于G2/M期,抑制肝癌细胞增殖,进而抑制肝癌生长.