【摘要】：目的:探討TGF-β1在大鼠睪丸支持細(xì)胞增殖和凋亡中的作用以及對(duì)緊密連接相關(guān)蛋白基因表達(dá)的影響。方法:體外分離和培養(yǎng)大鼠睪丸支持細(xì)胞,分空白對(duì)照組、TGF-β1受體阻滯劑組、TGF-β1刺激組和TGF-β1+受體阻滯劑組共4組。CCK-8檢測(cè)細(xì)胞增殖;流式細(xì)胞術(shù)檢測(cè)細(xì)胞凋亡;建立支持細(xì)胞原代雙室培養(yǎng)模型,應(yīng)用跨上皮電阻測(cè)定法(TER)檢測(cè)單層細(xì)胞兩側(cè)跨細(xì)胞電阻值;RT-PCR和Western印跡檢測(cè)支持細(xì)胞閉合蛋白(Occludin)、閉鎖小帶蛋白1(ZO-1)和緊密連接蛋白Ⅱ(ClaudinⅡ)的相對(duì)表達(dá)量。結(jié)果:各組細(xì)胞增殖組間差異有統(tǒng)計(jì)學(xué)意義(F=5.05,P0.05),TGF-β1刺激組與空白對(duì)照組相比細(xì)胞增殖率增加(P0.05),而TGF-β1+受體阻滯劑組與TGF-β1刺激組相比細(xì)胞增殖率降低(P0.05)。各組細(xì)胞凋亡率組間差異無(wú)統(tǒng)計(jì)學(xué)意義(F=1.13,P0.05);各組細(xì)胞間TER差異有統(tǒng)計(jì)學(xué)意義(F=38.99,P0.01),與空白對(duì)照組比較TGF-β1刺激組TER降低(P0.01),與TGF-β1刺激組比較,TGF-β1+受體阻滯劑組和TGF-β1受體阻滯劑組細(xì)胞TER升高(P均0.01);各組支持細(xì)胞中ZO-1、ClaudinⅡmRNA及蛋白在各組中的表達(dá)量差異均無(wú)統(tǒng)計(jì)學(xué)意義(F_(mRNA)=0.49,0.93,P均0.05;F_(蛋白)=0.28,1.31,P均0.05),而Occludin mRNA及蛋白在各組中的表達(dá)量差異均有統(tǒng)計(jì)學(xué)意義(F_(mRNA)=6.86,F_(蛋白)=6.87,P均0.05);與空白對(duì)照組相比,TGF-β1刺激組Occludin mRNA及蛋白表達(dá)明顯降低(P均0.01),而加入受體阻滯劑后,Occludin mRNA及蛋白表達(dá)明顯回升(P均0.05)。結(jié)論:TGF-β1對(duì)睪丸支持細(xì)胞的增殖有促進(jìn)作用,并能通過(guò)調(diào)節(jié)Occludin蛋白的表達(dá)而影響大鼠支持細(xì)胞間的緊密連接。
[Abstract]:Aim: to investigate the role of TGF- 尾 1 in proliferation and apoptosis of rat testicular Sertoli cells and the effect of TGF- 尾 1 on the expression of tight junction associated protein gene. Methods: rat testicular Sertoli cells were isolated and cultured in vitro. TGF- 尾 1 receptor blocker group and TGF- 尾 1 receptor blocker group were divided into 4 groups: CCK-8, TGF- 尾 1 receptor blocker group, flow cytometry to detect cell apoptosis, and TGF- 尾 1 receptor blocker group, TGF- 尾 1 receptor blocker group and TGF- 尾 1 receptor blocker group. The primary double compartment culture model of Sertoli cells was established. Transdermal resistance assay (TER) was used to detect the relative expression of closed cell protein (Occludin), (ZO-1) and tight junction protein 鈪，

本文編號(hào)：2190299