TGF-β1-SP1 Signaling Pathway Targets Regulating Guinea Pig Scleral Fibroblasts Secretion And Collagen Expression

Objective:To investigate the targeted regulation of TGFβ1-SP1 signaling pathway on the secretion and synthesis of collagen expression in guinea pig scleral fibroblasts.Methods:After primary culture of guinea pig scleral fibroblasts,immunohistochemical detection of Sp1 and typeⅠcollagen was performed.The protein and mRNA expressions of Sp1 and typeⅠcollagen were detected by RT-PCR and Western blot in four different groups of cells.Results:(1)After 5 days of tissue culture,the cell culture is completed,showing spindle-shaped and transparent,and the cells are flat and full.After three times of proliferation,the morphology was stable and arranged in an irregular fence.(2)Fibroblasts produce numerous vimentins in the cytoplasm.(3)Sp1 and type Ⅰ collagen are present in the cell tissues of all groups.(4)In the TGFβ1 group,the type Ⅰ collagen in the nucleus was stronger than the control group,and the difference was statistically significant(P<0.05);the TGFβ1+TGFβ1 inhibitor group was weaker than the control group,and the difference was statistically significant(P<0.05);The TGFβ1+Sp1 inhibitor group was stronger than the TGFβ1+TGFβ1 inhibitor group but weaker than the control group,and the difference was statistically significant(P<0.05).(5)The degree of coloration of Sp1 protein in the nucleus of the TGFβ1 group was significantly better than that of the control group,and thedifference was statistically significant(P<0.05);The difference was statistically significant(P<0.05);the TGFβ1+Sp1 inhibitor group was stronger than the TGFβ1+TGFβ1 inhibitor group,but was relatively weaker than the control group,and the difference was statistically significant(P<0.05).(6)Sp1 and type Ⅰ collagen m RNA were positive in guinea pig scleral fibroblasts.(7)The expression of Sp1 protein mRNA in the TGFβ1 group was stronger than that in the control group,and the difference was statistically significant(P<0.05);the TGFβ1+TGFβ1 inhibitor group was weaker than the control group,and the difference was statistically significant(P<0.05);TGFβ1 The+Sp1 inhibitor group was stronger than the TGFβ1+TGFβ1 inhibitor group but weaker than the control group,and the difference was statistically significant(P<0.05).(8)The expression of type Ⅰ collagen m RNA in the cells of the TGFβ1 group was stronger than that of the control group,and the difference was statistically significant(P<0.05);the TGFβ1+TGFβ1 inhibitor group was weaker than the control group,and the difference was statistically significant(P<0.05);The TGFβ1+Sp1 inhibitor group was stronger than the TGFβ1+TGFβ1 inhibitor group but weaker than the control group,and the difference was statistically significant(P<0.05).(9)In scleral fibroblasts,Sp1 and type Ⅰ collagen were positive.(10)The protein expression of Sp1 and type Ⅰ collagen in the TGFβ1 group was stronger than that in the control group,and the difference was statistically significant(P<0.05);the TGFβ1+TGFβ1 inhibitor group was weaker than the control group,and the difference was statistically significant(P<0.05);The TGFβ1+Sp1inhibitor group was stronger than the TGFβ1+TGFβ1 inhibitor group but weaker than the control group,and the difference was statistically significant(P<0.05).Conclusion:TGFβ1 can accelerate the expression of Sp1 protein and type Ⅰ collagen through the response,so it can effectively regulate the Sp1 signaling pathway,and then affect the expression of type Ⅰ collagen.