灯盏花素对晚期糖基化终产物诱导的大鼠系膜细胞的影响  

Influence of breviscapine on rat glomerular mesangial cells induced by advanced glycation end-products

作　　者：张满生[1] 张小云[1] 杨爱华[1] 李素波[1] 

机构地区：[1]杭州师范大学医学院附属萧山第一医院肾内科

出　　处：《中国临床药理学与治疗学》2016年第6期636-640,共5页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：杭州市科技局引导项目(2014)

摘　　要：目的:观察灯盏花素对晚期糖基化终产物(AGEs)诱导的大鼠肾小球系膜细胞(MC)增殖、氧化应激及转化生长因子β1(TGF-β1)、IV型胶原蛋白(ColⅣ)表达的影响。方法:采用体外细胞培养法,用不同浓度的灯盏花素(0、25、50、100、200 mg/L)和一定浓度的糖基化牛血清白蛋白(AGEs-BSA,100 mg/L)共培养大鼠MC细胞48h,相同条件下牛血清白蛋白(BSA)处理为对照,采用MTT法检测大鼠MC增殖,比色法检测培养上清液中氧化应激指标SOD、CAT、GSH-PX和丙二醛(MDA),ELISA法检测TGF-β1、ColⅣ浓度,RT-PCR法检测大鼠MC中TGF-β1mRNA的表达水平。结果:AGEs-BSA可以刺激大鼠MC细胞增殖,减少超氧化物歧化酶(SOD)、过氧化氢酶(CAT)及谷胱甘肽过氧化物酶(GSH-PX)活性,增加MDA含量,促进大鼠MC分泌TGF-β1及ColⅣ蛋白,上调大鼠MC中TGF-β1mRNA的表达(P均<0.05或0.01),而灯盏花素能以剂量依赖性抑制AGEs-BSA诱导的大鼠MC增殖,提高SOD、GSH-PX及CAT活性,减少MDA含量,减少大鼠MC TGF-β1和ColⅣ的分泌,下调大鼠TGF-β1mRNA的表达(P均<0.05或0.01)。结论:灯盏花素可以抑制AGEs-BSA诱导的大鼠MC增殖、氧化应激反应及TGF-β1和ColⅣ的表达,从而减少细胞外基质的合成,可能是其防治糖尿病肾病的机制之一。AIM： To study the influence of breviscapine on proliferation , oxidative stress state, expression of TGF-β1 and Col IV of rat glomerular mesangial cells （MC） induced by advanced glycation end-products （AGEs）. METHODS ： Rat glomerular MC were cultured for 48 hours and subjected to the treatment of AGEs-BSA （100mg/L） and breviscapine （0, 25, 50, 100, 200 mg/L） , while BSA addition was used as control. MTT method was used to measure cell proliferation, the activity of SOD, CAT, and GSH-PX , and MDA level in MC supernatant were detected by colorimety. The quantity of TGF-β1 and collagen IV in medium were determined by ELISA. The expression of TGF-β1 mRNA was analyzed by RT-PCR. RESULTS：AGEs could stimulate proliferation of MC, reduce the activity of SOD, CAT and GSH-PX, increase the level of MDA, promote the secretion of TGF-β 1and collagen IV in the medium , up-regulate the expression of TGF-β1 mRNA （ all P 〈 0.05 or 0.01 ） ; while breviscapine could inhibit the proliferation of MC, in- crease the activity of SOD, CAT, GSH-PX , decrease the level of MDA, reduce the secretion of TGF-β1 and Collagen IV in medium , down-regulate the expression of TGF-β1 mRNA iduced by AGEs all at a dose dependent manner （all P 〈 0.05 or 0. 01 ）. CONCLUSION： Breviscapine could inhibit AGEsinduced proliferation , oxidative stress state and the expression of TGF-β1 and collagen IV of rat glomerular MC, which could be one of the mechanisms of breviscapine preventing and treating diabetic nephropathy.

关 键 词：灯盏花素 晚期糖基化终产物 肾小球系膜细胞 细胞增殖 氧化应激 转化生长因子Β1 IV型胶原蛋白 

分 类 号：R965.2[医药卫生—药理学]