钙三醇抑制肾间质成纤维细胞的活化  

Effects of calcitriol on proliferation and extracellular matrix synthesis in renal interstitial fibroblasts

作　　者：王文姬[1] 王伟铭[1] 陈楠[1] 

机构地区：[1]上海交通大学医学院附属瑞金医院肾脏科,200025

出　　处：《中华肾脏病杂志》2008年第4期271-276,共6页Chinese Journal of Nephrology

基　　金：上海市重点学科（T0201）;上海市卫生局重点学科基金（05Ⅲ001）;上海市卫生局重点课题（2003ZD002）

摘　　要：目的通过观察钙三醇对肾间质成纤维细胞增殖和凋亡的影响，及其对转化生长因子（TGF）β1诱导的成纤维细胞转分化和细胞外基质（ECM）合成的干预作用，旨在探讨钙三醇抗肾间质纤维化的潜在效应及相关机制。方法体外培养大鼠肾间质成纤维细胞NRK-49F，分别以不同浓度TGF-β1（1、2、5和10μg／L）和（或）不同浓度钙三醇（10^-4、10^-5、10^-6、10^-7、和10^-8mmol／L）进行干扰。MTT法观察细胞增殖情况；流式细胞术分析细胞周期和细胞凋亡改变；实时定量PCR和Western印迹法分别检测细胞内α平滑肌肌动蛋白（d—SMA）、结缔组织生成因子（CTGF）及纤连蛋白（FN）的mRNA和蛋白表达。结果钙三醇能明显抑制正常和TGFβ1（5μg／L）诱导的NRK-49F细胞增殖（P〈0．01）。经不同浓度钙三醇（10^-4、10^-5、10^-6、10^-7、和10^-8mmol／L）作用48h后，NRK-49F细胞G2／S期细胞比例较TGF-β1刺激组均明显减少（分别为25．88％、21．81％、21．73％、23．28％、23．61％比27．42％，均P〈0．05），但对细胞凋亡无明显影响。NRK-49F细胞经TGF-β1刺激后，其α-SMA、CTGF、FNmRNA表达量显著上升，并在TGF-β1～5μg／L的范围内呈剂量依赖效应。钙三醇能明显抑制TGF-β1诱导的α-SMA、CTGF、FNmRNA表达上调（均P〈0．05），而不同钙三醇浓度干预组之间差异无统计学意义。钙三醇能显著降低TGF-β1诱导的α-SMA和FN蛋白表达（P〈0．05）。结论钙三醇可通过G1期停滞抑制大鼠肾间质成纤维细胞增殖，但不影响凋亡。钙三醇具有拈抗TGF-β1致肾间质纤维化的潜在作用。Objective To observe the effects of calcitriol on cellular proliferation and apoptosis, and on the expression of transforming growth factor-β1 （TGF-β1）-induced α-SMA and extracellular matrix （ECM） synthesis in renal interstitial fibroblasts, so as to illustrate its potential roles and underlying mechanisms in preventing renal interstitial fibrosis. Methods In NRK- 49F of cultured rats in vitro, cellular proliferation was detected by MTT. The cellular cycle and apoptosis were assessed by flow cytometry using annexin-V FITC/propidium iodide （PI）. Real-time PCR was performed to detect the mRNA expression of α-SMA, CTGF and FN, and Western blot was performed to detect the protein expression of α-SMA and FN. Results Cellular proliferation was inhibited by calcitriol through Gl-arrest （P〈0.05）. But calcitriol had no effect on cell apoptosis （P〉0.05）. Increased expression of α-SMA, CTGF and FN mRNA induced by TGF-β1 were significantly suppressed by calcitriol as compared with TGF-β1 treated group （P〈0.05）. However, there was no significant difference among the groups with various concentration of calcitrol （P〉0.05）. The similar phenomenon of the level of α-SMA and FN protein expression was observed in all groups by Western blot. Conclusion Calcitriol inhibits cellular proliferation of renal interstitial fibroblast by Gl-arrest, but has no effect on cell apoptosis, and can prevent renal interstitial fibrosis caused by TGF-β1.

关 键 词：骨化三醇 成纤维细胞 转化生长因子Β 细胞增殖 细胞外基质 

分 类 号：R692[医药卫生—泌尿科学] R692.5[医药卫生—外科学]