摘要
目的:基于TGF-β1/Smads通路探讨附子水提物及附子多糖对肾脏纤维化的影响及可能作用机制。方法:体外培养肾小球内皮细胞(HRGECs),采用TGF-β1刺激诱导HRGECs纤维化样模型。造模成功后将细胞分为正常组、TGF-β1模型组(30 ng/mL)、附子水提物组(10 mg/mL)、附子多糖组(5 mg/mL)。光学显微镜、扫描电镜观察细胞形态的变化;细胞划痕实验、Transwell实验观察细胞迁移和侵袭能力;RT-qPCR法检测α-SMA、FSP-1、ZO-1、CD31 mRNA表达水平;Western-blot法检测α-SMA、FSP-1、ZO-1、CD31、p-Smad3、Smad7蛋白表达水平。结果:与正常组比较,TGF-β1刺激72 h后HRGECs形态出现纤维化样变,迁移和侵袭能力增强(P<0.01),α-SMA、FSP-1蛋白及mRNA表达升高,ZO-1、CD31蛋白及mRNA表达下调(P<0.05,P<0.01)。与TGF-β1模型组比较,附子水提物组、附子多糖组细胞形态改善,迁移和侵袭能力降低(P<0.01),α-SMA、FSP-1、p-Smad3蛋白及mRNA表达下降;ZO-1、CD31、Smad7蛋白及mRNA表达升高(P<0.05,P<0.01)。结论:TGF-β1可明显诱导肾小球内皮细胞发生纤维化改变。附子水提物及附子多糖对TGF-β1诱导的HRGECs纤维化改变有明显的改善作用,其机制可能与调节TGF-β1/Smads信号通路有关。
Objective:Based on the signal pathway of TGF-β/Smads,to explore the effects and mechanisms of Aconiti Lateralis Radix Praeparata(ALRP)and its polysaccharide on renal fibrosis.Methods:HRGECs were cultured in vitro,and TGF-β1 was used to stimulate and induce fibrosis model of HRGECs.After successful modeling,the cells were divided into control group,TGF-β1 group(30 ng/mL),ALRP group(10 mg/mL)and polysaccharide group(5 mg/mL).The morphological changes of cells were observed by optical microscope and scanning electron microscope.Cell migration and invasion ability were observed by cell scratch assay and Transwell assay.RT-qPCR was applied to detect mRNA levels ofα-SMA,FSP-1,Zo-1 and CD31.The protein expression levels ofα-SMA,FSP-1,Zo-1,CD31,p-Smad3 and Smad7 were detected by Western blot.Results:Compared with control group,after 72 h stimulation with TGF-β1,the morphology of HRGECs was fibrotic,migration and invasion ability was enhanced(P<0.01),the protein and mRNA expressions ofα-SMA and FSP-1 increased,and the expression of Zo-1 and CD31 protein and mRNA were down-regulated(P<0.05 or P<0.01).Compared with TGF-β1 group,the cell morphology of ALRP group and polysaccharide group was improved,migration and invasion ability decreased(P<0.01),and the protein and mRNA expressions ofα-SMA,FSP-1 and p-Smad3 decreased.The protein and mRNA expressions of Zo-1,CD31 and Smad7 increased(P<0.05 or P<0.01).Conclusion:TGF-β1 can clearly induce fibrotic changes in glomerular endothelial cells.Water extract of aconiti lateralis radix praeparata and its polysaccharide of have obvious inhibitory effect on TGF-β1-induced HRGECs fibrosis,inhibit cell migration and invasion,which may be related to the regulation of TGF-β/Smads signaling pathway.
作者
赵延红
李星瑶
姜甜甜
高琪
叶冰玉
安鹏
石兴民
蔡子墨
吴喜利
ZHAO Yanhong;LI Xingyao;JIANG Tiantian;GAO Qi;YE Bingyu;AN Peng;SHI Xingmin;CAI Zimo;WU Xili(Department of Integrated Traditional Chinese and Western Medicine,The Second Aliated Hospital of Xi'an Jiaotong University,Xi'an 710004 China;Department of Rheumatism,Xi'an Fifth Hospital,Xi'an 710082 China;Medical Department of Xi'an Jiaotong University,Xi'an 710061 China)
出处
《中医药学报》
CAS
2024年第11期29-35,共7页
Acta Chinese Medicine and Pharmacology
基金
中医药“双链融合”中青年创新团队(2022-SLRH-LJ-011)
秦创原中药创新研发项目(2022-QCYZH-019)。
