【摘要】：目的:探討TGF-β1在大鼠睪丸支持細胞增殖和凋亡中的作用以及對緊密連接相關蛋白基因表達的影響。方法:體外分離和培養(yǎng)大鼠睪丸支持細胞,分空白對照組、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檢測細胞增殖;流式細胞術檢測細胞凋亡;建立支持細胞原代雙室培養(yǎng)模型,應用跨上皮電阻測定法(TER)檢測單層細胞兩側跨細胞電阻值;RT-PCR和Western印跡檢測支持細胞閉合蛋白(Occludin)、閉鎖小帶蛋白1(ZO-1)和緊密連接蛋白Ⅱ(ClaudinⅡ)的相對表達量。結果:各組細胞增殖組間差異有統(tǒng)計學意義(F=5.05,P0.05),TGF-β1刺激組與空白對照組相比細胞增殖率增加(P0.05),而TGF-β1+受體阻滯劑組與TGF-β1刺激組相比細胞增殖率降低(P0.05)。各組細胞凋亡率組間差異無統(tǒng)計學意義(F=1.13,P0.05);各組細胞間TER差異有統(tǒng)計學意義(F=38.99,P0.01),與空白對照組比較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受體阻滯劑組細胞TER升高(P均0.01);各組支持細胞中ZO-1、ClaudinⅡmRNA及蛋白在各組中的表達量差異均無統(tǒng)計學意義(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量差異均有統(tǒng)計學意義(F_(mRNA)=6.86,F_(蛋白)=6.87,P均0.05);與空白對照組相比,TGF-β1刺激組Occludin mRNA及蛋白表達明顯降低(P均0.01),而加入受體阻滯劑后,Occludin mRNA及蛋白表達明顯回升(P均0.05)。結論:TGF-β1對睪丸支持細胞的增殖有促進作用,并能通過調(diào)節(jié)Occludin蛋白的表達而影響大鼠支持細胞間的緊密連接。
[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，

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