熊果酸对大鼠肾小球系膜细胞凋亡及TGF-β1/Smads信号通路的影响  

Effect of ursolic acid on the apoptosis and TGF-β/Smads signal pathway of cultured rat mesangial cells

作　　者：林志民[1] 施书涵[1] 李月婷[1] 郑兴中[2] 黄荣桂[1] LIN Zhi-min;SHI Shu-han;LI Yue-ting;ZHENG Xing-zhong;HUANG Rong-gui(Department of Nephrology,the Second Affiliated Hospital of Fujian Medical University,Fujian Quanzhou 362000,China;Department of Pharmacy,the Second Affiliated Hospital of Fujian Medical University,Fujian Quanzhou 362000,China)

机构地区：[1]福建医科大学附属第二医院,肾内科,福建泉州362000 [2]福建医科大学附属第二医院,药学部,福建泉州362000

出　　处：《中国医院药学杂志》2020年第18期1929-1931,1978,共4页Chinese Journal of Hospital Pharmacy

基　　金：福建省自然科学基金项目(编号:2016J01523)。

摘　　要：目的:观察熊果酸(ursolic acid,UA)对活化的大鼠肾小球系膜细胞(mesangial cells,MCs)凋亡与TGF-β1/smads信号通路的影响,探讨UA抗肾纤维化的作用机制。方法:原代培养并分离、纯化大鼠肾小球系膜细胞,转化生长因子β1(TGF-β1)刺激活化后,熊果酸按低剂量(2.5μmol·L-1)、中剂量(5μmol·L-1)、高剂量(10μmol·L-1)进行干预。Cell counting kit-8(CCK-8)及流式细胞仪分别检测系膜细胞活力及凋亡率,免疫印迹法测定系膜细胞TGF-β1受体(TβRI、TβRII)、磷酸化蛋白P-smad信号分子表达的变化。结果:5μg·L-1 TGF-β1明显激活系膜细胞增殖,上调TGF-β1受体表达及Smad2的磷酸化,抑制系膜细胞凋亡。低、中、高剂量熊果酸均能抑制系膜细胞增殖,促进凋亡,并下调系膜细胞TGF-β1受体表达,抑制Smad2的磷酸化。结论:熊果酸可通过影响TGF-β1/Smads信号通路,诱导系膜细胞凋亡,拮抗细胞增殖,这可能是熊果酸抗肾脏纤维化的部分细胞学机制。OBJECTIVE To observe the effects of ursolic acid(UA)on apoptosis and TGF-β1/smads signaling pathway of activated rat glomerular cells(MCs),and to explore the mechanism of UA anti-renal fibrosis.METHODS Rat glomerular cells were cultured,isolated and purified.After activation was stimulated by transforming growth factorβ1(TGFβ1),ursolic acid was intervened at low dose(2.5μmol·L-1),medium dose(5μmol·L-1)and high dose(10μmol·L-1).Cell counting kit-8(CCK-8)and flow cytometry were used to detect the viability and apoptosis rate of cells,respectively.The expression of TGF-β1 receptor(TβRI,TβRII)and phosphorylated protein P-smad signaling molecules in cells was determined by immunoblotting.RESULTS 5μg·L-1 TGF-β1 significantly activated cell proliferation,up-regulated TGF-β1 receptor expression and Smad2 phosphorylation,and inhibited apoptosis.Low,medium and high doses of ursolic acid could inhibit cell proliferation,promote apoptosis,down-regulate TGF-β1 receptor expression and inhibit the phosphorylation of Smad2.CONCLUSION Ursolic acid can antagonize mesangial cell proliferation by promoting apoptosis of mesangial cells and inhibiting the TGF-β1/Smads signaling pathway,which may be part of the cytological mechanism of ursolic acid against renal fibrosis.

关 键 词：熊果酸 大鼠 肾小球系膜细胞 凋亡 TGF-β1/Smads信号通路 

分 类 号：R285[医药卫生—中药学]