高压诱导大鼠系膜细胞细胞外基质积聚机制及阿托伐他汀的干预作用  

Mechanism of high pressure-induced extracellular matrix accumulation in mesangial cells and the interventional effect of atorvastatin

作　　者：董一飞[1] 程晓曙[1] 杨人强[1] 沈云峰[1] 徐劲松[1] 

机构地区：[1]江西医学院心血管研究所江西医学院第二附属医院心内科,南昌330006

出　　处：《中华肾脏病杂志》2005年第5期274-277,共4页Chinese Journal of Nephrology

基　　金：国家自然科学基金(30260037)

摘　　要：目的探讨高压诱导大鼠系膜细胞细胞外基质(ECM)积聚的可能途径,并观察阿托伐他汀的干预作用。方法将8～15代大鼠系膜细胞置于80mmHg高压下培养6、12、24、48h。并进一步分为阿托伐他汀组、TGF—β1反义寡核苷酸组、助溶剂二甲基亚砜(DMSO)对照组、错义寡核苷酸对照组及正常对照组。放射免疫法测定AngⅡ浓度。RT—PCR法测定TGF—β1、胶原纤维(Col)Ⅰ(α1)及纤溶酶原激活物抑制剂(PAI)-1mRNA表达量。Western印迹法测定上述各指标蛋白表达。结果阿托伐他汀能显著降低AngⅡ水平。TGF—β1反义寡核苷酸对AngⅡ水平无显著影响。阿托伐他汀和TGF-β1反义寡核苷酸均能明显降低TGF—β1、colⅠ(α1)及PAI-1mRNA和蛋白水平。错义寡核苷酸和二甲基亚砜对上述指标表达水平无显著影响。结论AngⅡ和TGF—β1参与高压诱导ECM的积聚过程。AngⅡ的作用由TGF—β1介导实现,即存在“压力→AngⅡ→TGF-β1→ECM积聚”作用途径。阿托伐他汀在一定程度上逆转高压诱导的ECM积聚,此作用可通过干预上述途径实现。Objective To explore the possible way of high pressure-induced extracellular matrix (ECM) accumulation in rat mesangial cells, and to observe the interventional effect of atorvaslatin. Methods According to different interventional factors, mesangial cells were divided into five groups: atorvastatin, dimethyl sulphoxide (DMSO), transforming growth factor-pi antisense oligodeoxynucleotides (TGF-β1 ASDON), scrambled oligodeoxynucleotides (SODN) and control group. All these groups were further exposed to the pressure of 80 mmHg for 6, 12, 24 and 48 hours respectively. The concentrations of angiotensin II in culture mediums were assessed by 125I-Angiotensin Radioimmunoassay Kit. Semi-quantitative RT-PCR and Western blotting were applied to examine the expression of TGF-β1, Collagen I (α1) and PAI-1 respectively at mRNA and protein level. Results Atorvastatin could significantly decrease the concentration of Ang II compared to the control group, while no statistical difference of Ang II expression was found between TGF-β1 ASODN group and control group. Compared to the control, atorvastatin and TGF-β1 ASODN could extensively inhibit the expression of mRNA and protein for TGF-β1, Collagen I (α1) and PAI-1. While no differences of the expression of above factors were found in scrambled oligodeoxynucleotide and DMSO groups as compared with the control group. Conclusions Ang II mediated by TGF-β1 is involved in the pressure-induced ECM accumulation through 'pressure→Ang II→TGF-β→ECM accumulation'chain. Atorvastatin can reverse the effect of pressure-induced ECM accumulation in mesangial cells to some extent, by interfering the above mentioned chain.

关 键 词：阿托伐他汀 系膜细胞 干预作用 诱导大鼠 细胞外基质积聚 高压 二甲基亚砜(DMSO) 细胞外基质(ECM) 纤溶酶原激活物抑制剂 WESTERN印迹法 TGF-β1 反义寡核苷酸 错义寡核苷酸 放射免疫法测定 RT-PCR法 mRNA表达量 AngⅡ 

分 类 号：R692[医药卫生—泌尿科学]