阿托伐他汀通过p38丝氨酸蛋白激酶信号通路干预人肾小球系膜细胞增殖和转化生长因子β1的表达(英文)  

Atorvastatin inhibits cell proliferation and decreases the expression of transforming growth factor-beta1 via p38 mitogen-activated protein kinases-pathway in human gomerular mesangial cells

作　　者：李萍[1] 张海燕[1] 刘国良[1] 盛志业[2] 马小峰[3] 向军[4] 

机构地区：[1]中国医科大学附属第一医院内分泌科,辽宁省沈阳市110001 [2]解放军沈阳军区总医院内分泌科,辽宁省沈阳市110016 [3]解放军沈阳军区总医院呼吸科,辽宁省沈阳市110016 [4]解放军沈阳军区总医院泌尿外科,辽宁省沈阳市110016

出　　处：《中国组织工程研究与临床康复》2007年第16期3184-3188,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：辽宁省科技攻关项目(2001225004)~~

摘　　要：背景:p38丝氨酸蛋白激酶信号激活引起细胞生长、增殖、分化,可能是糖尿病血管并发症发生发展的共同通路。目的:观察阿托伐他汀对抗p38丝氨酸蛋白激酶信号通路的激活,及其对氧化修饰低密度脂蛋白作用下人肾小球系膜细胞增殖与转化生长因子β1表达的干预。设计:随机、平行对照、开放性实验。单位:中国医科大学附属第一医院内分泌科,解放军沈阳军区总医院内分泌科、呼吸科及泌尿外科。材料:实验于2004-05/2005-05先后在中国医科大学药理教研室、沈阳军区总医院呼吸科实验室完成。以肾肿瘤行单侧肾切除患者正常部分的肾皮质(由沈阳军区总医院泌尿外科向军主任提供,征得患者同意)作为实验用人肾小球系膜细胞的来源。氧化修饰低密度脂蛋白浓度为(5.3±1.0)nmol/100μg(购自协和医科大学生化研究所,批号20040711);阿托伐他汀(辉瑞制药,批号45837088);p38丝氨酸蛋白激酶单克隆抗体(Santa Cruz)。方法:①取肾肿瘤行单侧肾切除患者的正常部分肾皮质6.0～8.0cm3,分离制备肾小球系膜细胞,待生长单层达80%培养面积后进行消化传代。传至第2代时,观察细胞形态,行DAB染色,胞浆内出现棕黄色团块或泥沙样颗粒为DAB染色阳性。油红“O”染色检测细胞吞噬氧化修饰低密度脂蛋白的情况。取传至4～10代的细胞用于实验。②按5×103个细胞接种于96孔板,200μL/孔。实验共分5组:阿托伐他汀1.2,6,12mg/L浓度组分别加入对应浓度的阿托伐他汀溶液,氧化低密度脂蛋白组和空白对照组此时仅加入培养基,6个平行孔/组。各组预处理30min后,除空白对照组外均暴露于终浓度为80mg/L的氧化修饰低密度脂蛋白中,24h后每孔加入四甲基偶氮唑蓝,在492nm波长的酶标仪上检测吸光度值,计算细胞增殖抑制率。③将1×106～3×106细胞接种于6个200mL培养瓶中,随机数字表法选1瓶作为空白对照组,第2～4�BACKGROUND： The cell growth, proliferation and differentiation caused by p38 mitogen-activated protein kinase （p38MAPK） might act as the common pathway in the onset and development of diabetic vascular complication. OBJECTIVE： To investigate the effect of atorvastatin on p38MAPK signal pathway and the influence of atorvastatin on cell proliferation and expression of transforming growth factor-β1 （TGF-β1） at transcriptional level in human glomerular mesangial cells （HGMCs） cultured with oxidative modification of low-density lipoprotein （ox-LDL）. DESIGN ： A randomized, parallelized, controlled and open tria. SETTING： Endocrinology Department, First Hospital Affiliated to China Medical University; Endocrinology Department, Respiratory Department, Urology Department, the General Hospital of Shenyang Military Area Command of Chinese PLA. MATERIALS： The experiment had been done in the laboratories for Pharmaceutical Department of China Medical University and Respiratory Department of Shenyang Military Area Command of Chinese PLA from May 2004 to May 2005. The sample was cut from renal cortex from the healthy segment of nephroectomy from a tumor patient （Provided by Xiang Jun, Urology Department, the General Hospital of Shenyang Military Area Command of Chinese PLA; Informed consent was obtained）. OX-LDL was purchased from biochemistry institute of Peking Union Medical College （Batch No. 20040711 ）. ox-LDL was 5.3±1.0 nmol in 100μg protein. Atorvastatin was purchased from Pfizer Pharmaceutical Co. Ltd （No. 45837088）; p38MAPK monocloncal antibody was purchased from Santa Cruz. METHODS ： （1） 6.0-8.0 cm^3 blocks of cortex were cut from renal cortex from the healthy segment of nephroectomy from a tumor patient, glomerular mesangial cells were isolated. When grew into 80% confluent monolayer, the cells were digested and performed passage. After the first two passages, the cells were pure based on morphology and characterized by DAB staining for Vimentin antigen and act

关 键 词：庚酸类/药理学 吡咯类/药理学 细胞增殖 转化生长因子β 丝裂原激活蛋白激酶类 肾小球膜 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]