TGFβ1-Sp1信号通路靶向调控豚鼠巩膜成纤维细胞分泌与合成胶原表达的研究

TGFβ1-Sp1 signaling pathway targets regulating secretion and synthetic collagen expression in guinea pig scleral fibroblasts

目的研究转化生长因子β1(TGFβ1)通过特异性蛋白1(Sp1)信号传导通路调控Ⅰ型胶原合成与降解的影响。方法通过豚鼠巩膜成纤维细胞的原代培养后,进行免疫组织化学染色体外检测Sp1和Ⅰ型胶原的表达。分别用不同处理的4组细胞做qRT-PCR及Western blot法检测Sp1与Ⅰ型胶原的蛋白和mRNA表达。结果与对照组相比,Sp1蛋白及Ⅰ型胶原蛋白的表达量和mRNA量在TGFβ1组中均表达增强(P<0.05)、TGFβ1+TGFβ1抑制剂组表达均减弱(P<0.05)、TGFβ1+Sp1抑制剂组中的表达弱但比TGFβ1+TGFβ1抑制剂组强(P<0.05)。结论TGFβ1可以促进Sp1蛋白和Ⅰ型胶原的表达,从而认为TGFβ1可以通过Sp1信号传导通路调控Ⅰ型胶原合成与降解。

Objective To investigate the effects of TGFβ1 on the synthesis and degradation of type Ⅰ collagen through the Sp1 signaling pathway. Methods After primary culture of guinea pig scleral fibroblasts, immunohistochemical extrachromosomal detection of Sp1 and type Ⅰ collagen was performed. The mRNA and protein expression of type Ⅰ collagen was detected by qRT-PCR and Western blot method in 4 groups of cells with different treatments. Results Compared with the control group,the expressions of Sp1 protein,type Ⅰ collagen and mRNA increased in the TGFβ1 group(P<0.05),and the expressions in TGFβ1+TGFβ1 inhibitor group were weakened(P<0.05),TGFβ1.The expressions in the +Sp1 inhibitor group were weaker but stronger than those in the TGFβ1+TGFβ1 inhibitor group(P<0.05).Conclusion TGFβ1 can promote the expression of Sp1 protein and type Ⅰ collagen,and thus TGFβ1 can regulate the synthesis and degradation of type Ⅰ collagen through Sp1 signaling pathway.