乌索酸对高糖诱导人系膜细胞损伤的作用  

Effects of Ursolic Acid on High Glucose-induced Injury in Human Mesangial Cells

作　　者：岳媛[1,2] 范秋灵[1] 都姝妍[3] 远方[2] 徐莉[1] 李琳[1] 刘楠[3] 姜奕[3] 王力宁[1] YUE Yuan;FAN Qiuling;DU Shuyan;YUAN Fang;XU Li;LI Lin;LIU Nan;JIANG Yi;WANG Lining(Department of Nephrology,The First Hospital,China Medical University,Shenyang 110001,China;Department of Nephrology,The Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110032,China;Central Laboratory,The First Hospital,China Medical Uni-versity,Shenyang 110001,China)

机构地区：[1]中国医科大学附属第一医院肾内科,沈阳110001 [2]辽宁中医药大学附属医院肾内科,沈阳110032 [3]中国医科大学附属第一医院中心实验室,沈阳110001

出　　处：《中国医科大学学报》2018年第8期682-686,共5页Journal of China Medical University

基　　金：国家自然科学基金(81770724;81270808);辽宁省自然科学基金(201602821);沈阳市科技计划(F16-205-1-40)

摘　　要：目的探讨乌索酸(UA)对高糖诱导人系膜细胞损伤的保护作用。方法人系膜细胞复苏后,用MCM 4201完全培养基常规培养,置于37℃、饱和湿度5%CO_2的孵箱内贴壁传代培养,隔天换液1次,取5~9代对数生长期细胞,分为正常糖组(5.5mmol·L^(-1)葡萄糖),高糖组(30.0 mmol·L^(-1)葡萄糖),高渗对照组(5.5 mmol·L^(-1)葡萄糖+24.5 mmol·L^(-1)甘露醇),乌索酸治疗A、B、C组(30.0 mmol·L^(-1)葡萄糖,A、B、C组分别给予0.5、1.0、2.0μmol·L^(-1)乌索酸)。6组均给药48 h。利用MTT比色法检测细胞增殖,流式细胞仪检测系膜细胞凋亡,采用实时PCR及Western blotting检测细胞凋亡相关蛋白Bcl-x L、Bax、survivin m RNA及蛋白表达,细胞损伤相关蛋白TGF-β1、FN m RNA及蛋白表达。结果 0.5μmol·L^(-1)乌索酸治疗组与正常糖组、高渗对照组系膜细胞增殖程度差异无统计学意义(P>0.05),2.0μmol·L^(-1)乌索酸治疗组大部分系膜细胞已经死亡。1.0μmol·L^(-1)乌索酸组抑制高糖诱导的系膜细胞增殖和TGF-β1、FN、Bcl-xl、survivin m RNA及蛋白表达,增强促凋亡基因Bax m RNA及蛋白表达,促进系膜细凋亡。结论乌索酸通过促进系膜细胞早期凋亡抑制系膜细胞增殖,抑制TGF-β1表达和FN聚集,减轻系膜细胞损伤。Objective To investigate the effect of ursolic acid（UA） on apoptosis in human renal mesangial cells（RMCs） cultured in a high-glucose medium. Methods After the recovery of RMCs cultured in MCM 4201 medium at 37 ℃ and 5% CO2,with media being replenished every alternate day,5-9 generations of cells in the logarithmic phase of growth were selected and divided into a normal glucose group（5.5 mmol·L^-1 glucose）,high glucose group（30.0 mmol·L^-1 glucose）,high osmotic group（5.5 mmol·L^-1 glucose ＋24.5 mmol·L^-1 mannitol）,and treatment A group（30.0 mmol·L^-1 glucose＋0.5 μmol·L^-1 ursolic acid）,treatment B group（30.0 mmol·L^-1 glucose＋1.0 μmol·L^-1 ursolic acid）,treatment C group（30.0 mmol·L^-1 glucose＋2.0 μmol·L^-1 ursolic acid）. After 48 h of culturing,RMC proliferation was assessed using an MTT assay;apoptosis was assessed via Annexin V-FITC double staining flow cytometry. The expression of Bcl-xl,Bax,survivin,TGF-β1,and FN proteins and m RNA were analyzed via Western blotting and RT-PCR,respectively. Results UA at 1.0 μmol·L^-1 suppressed RMC proliferation and TGF-β1,FN,Bcl-xl,and survivin m RNA and protein expression induced by HG. UA at 1.0 μmol·L^-1 promoted apoptosis in RMCs by upregulating Bax m RNA and protein. Conclusion UA can reduce mesangial cell injury by promoting early apoptosis,inhibiting proliferation,downregulating TGF-β1,and FN aggregation.

关 键 词：人系膜细胞 高糖 乌索酸 细胞凋亡 

分 类 号：R320.2[医药卫生—人体解剖和组织胚胎学]