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目的:观察熊果酸(ursolic acid,UA)含药血清对体外脂多糖(LPS)刺激下大鼠肾小球系膜细胞(mesangial cells,MCs)增殖与转化生长因子β1(TGF-β1)表达的影响,探讨其保护肾脏的作用机制。方法:体外原代培养并鉴定大鼠肾小球系膜细胞;熊果酸含药血清分为3个剂量干预组,按高剂量(100 mg·kg~(-1))、中剂量(50 mg·kg~(-1))、低剂量(25 mg·kg~(-1))每日1次连续给大鼠灌胃,7 d后取血制备含药血清;正常对照组灌胃同体积生理盐水制备正常血清。系膜细胞用LPS(10mg·L~(-1))诱导后,分别加入正常血清对照或不同剂量熊果酸含药血清培养48 h。采用CCK-8法观察系膜细胞增殖,ELISA法检测细胞分泌TGF-β1水平,RT-PCR测定TGF-β1 mRNA的表达。结果:LPS明显诱导系膜细胞增殖,上调TGF-β1的表达。高、中、低剂量熊果酸含药血清均能抑制LPS刺激下系膜细胞增殖,并在基因水平呈剂量依赖性抑制TGF-β1的表达。结论:熊果酸含药血清可抑制炎症状态下系膜细胞增殖,并下调TGF-β1的表达,提示熊果酸对肾小球疾病可能发挥保护作用。
OBJECTIVE: To observe the effects of ursolic acid (UA) containing serum on the proliferation and the expression of transforming growth factor-β1 (TGF-β1) in rat mesangial cells (MCs) stimulated by lipopolysaccharide (LPS) To explore its mechanism of protecting kidneys. Methods: Rat mesangial cells were cultured and identified in primary culture. UA serum was divided into 3 groups: high dose (100 mg · kg -1), middle dose (50 mg · kg -1) The rats in the low dose (25 mg · kg -1) group were given gavage once a day for 7 days, then the serum was prepared by blood sampling. The normal control group was given gavage Normal saline was prepared by volume of normal saline. After induced by LPS (10 mg · L -1), mesangial cells were added to normal serum control or different doses of ursolic acid-containing serum for 48 h. The proliferation of mesangial cells was observed by CCK-8 assay. The levels of TGF-β1 secreted by cells were measured by ELISA. The expression of TGF-β1 mRNA was detected by RT-PCR. Results: LPS significantly induced mesangial cell proliferation and up-regulated the expression of TGF-β1. High, medium and low doses of ursolic acid-containing serum can inhibit LPS stimulation of mesangial cell proliferation, and a gene dose-dependent inhibition of TGF-β1 expression. Conclusion: The ursolic acid-containing serum can inhibit the proliferation of mesangial cells in inflammatory state and down-regulate the expression of TGF-β1, suggesting that ursolic acid might play a protective role in glomerular diseases.