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目的:探讨芪藤消浊颗粒对脂多糖(LPS)诱导的小鼠肾小球系膜细胞(MMCs)炎症和增殖的影响。方法:以3μg/mL LPS诱导小鼠肾小球系膜细胞构建慢性肾小球肾炎体外模型。采用Western blot法和RT-qPCR法检测ALKBH5和TRIM13的表达；通过ELISA法测定MMCs上清液中炎症因子白细胞介素-1β(IL-1β)、IL-10、肿瘤坏死因子-α(TNF-α)的含量；甲基化RNA免疫共沉淀法检测Trim13 mRNA的甲基化水平；流式细胞术分析细胞周期与细胞凋亡情况；CCK-8法检测细胞的增殖活性。结果:与正常对照组比较，模型对照组MMCs细胞炎症因子IL-1β、TNF-α含量显著升高，IL-10含量显著降低(P<0.01),P65、p-P65 NF-κB显著上调(P<0.01),G1/G0期细胞水平显著增加(P<0.01),细胞增殖活性显著升高(P<0.01),ALKBH5蛋白表达显著上调(P<0.01),Trim13 mRNA甲基化表达显著下调(P<0.01);与模型对照组比较，芪藤消浊颗粒20%含药血清组炎症因子IL-1β、TNF-α含量显著降低，IL-10含量显著升高(P<0.01),P65、p-P65 NF-κB蛋白表达显著下调(P<0.01),G1/G0期细胞水平显著减少(P<0.01),细胞增殖活性显著降低(P<0.01),ALKBH5蛋白表达显著下调，Trim13 mRNA甲基化表达显著上调(P<0.01)。ALKBH5过表达的MMCs中，芪藤消浊颗粒20%含药血清对细胞凋亡和细胞周期停滞的促进作用显著减弱(P<0.01)。结论:芪藤消浊颗粒通过调节ALKBH5-TRIM13轴，减轻MMCs的炎症反应，抑制其过度增殖，发挥治疗慢性肾小球肾炎的作用。
Abstract：Objective:To investigate the effects of Qiteng Xiaozhuo(芪藤消浊) Granules(QTXZG) on lipopolysaccharide(LPS)-induced inflammation and proliferation of mouse glomerular mesangial cells. Methods:Mouse glomerular mesangial cells(MMCs) were treated with 3 μg/mL LPS for the modeling of glomerulonephritis in vitro. Western blot and RT-qPCR were utilized to determine the protein and mRNA levels, respectively, of AlkB homolog 5(ALKBH5) and tripartite motif-containing 13(TRIM13). ELISA was used to measure the levels of interleukin-1β(IL-1β),tumor necrosis factor-alpha(TNF-α),and interleukin-10(IL-10) in the supernatant of MMCs. Methylated RNA immunoprecipitation was employed to assess the methylation level of Trim13 mRNA. Flow cytometry was adopted to analyze the cell cycle and apoptosis, while the CCK-8 assay was used to assess the cell proliferation. Results:Compared with the normal control group, the model control group showed elevations in IL-1β and TNF-α levels and a decline in IL-10 level(P<0.01),enhanced activation of the P65 NF-κB signaling pathway(P<0.01),increased G1/0 phase cells(P<0.01),enhanced cell proliferation(P<0.01),up-regulated ALKBH5 expression(P<0.01),and down-regulated methylation of Trim13 mRNA(P<0.01). Compared with the model control group, the QTXZG-containing serum lowered the IL-1β and TNF-α levels and raised the IL-10 level(P<0.01),inhibited the activation of the P65 NF-κB signaling pathway(P<0.01),reduced the G1/0 phase cells(P<0.01),weakened the cell proliferation(P<0.01),down-regulated the ALKBH5 expression, and up-regulated the methylation of Trim13 mRNA(P<0.01). In the MMCs with overexpression of ALKBH5,the promotion of cell apoptosis and cell cycle arrest by the QTXZG-containing serum was attenuated(P<0.01). Conclusion:QTXZG exerts a therapeutic effect on chronic glomerulonephritis by regulating the ALKBH5-TRIM13 axis to relieve the inflammation and inhibit the excessive proliferation of MMCs.
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基本信息:
DOI：10.13412/j.cnki.zyyl.20250605.002
中图分类号:R285.5
引用信息:
[1]雷淑军,高家荣,胡兴民,等.芪藤消浊颗粒通过ALKBH5调控Trim13 mRNA 甲基化减轻LPS诱导的肾小球系膜细胞炎症增殖[J].中药药理与临床,2026,42(02):43-50.DOI:10.13412/j.cnki.zyyl.20250605.002.
基金信息:
安徽省高等学校科学研究重点项目(编号：2022AH050455);安徽省高等学校科学研究重点项目(编号：2024AH040166)
2025-06-06
2025-06-06
2025-06-06