| Objectives:To investigate the regulatory mechanism of Rgl,which is the main component of Panax notoginseng,on the TGF-β/Smad signaling pathway in diabetic retinopathy.Through the investigate of Rgl treated high glucose induced human retinal vascular endothelial cells in the expression of TGF-β,smad2/3 and other related fibrosis factors,revealed the mechanism of Rgl intervention the development of diabetic retinopathy and reduce retinal fibrosis,which provide molecular theoretical foundation and new drug selection for the treatment of DR.Methods:HRMECs in human retinal endothelial cells were cultured in vitro,and the fourth generation cells were identified.Experimental group:(1)Normal control group:Human retinal vascular endothelial cells cultured in vitro with conventional D-glucose(5.5 mM);(2)Hyperglycemic group:human retinal vascular endothelial cells cultured in vitro with high concentration of D-glucose(30mM);(3)Rgl intervention group:human retinal vascular endothelial cells were cultured at a high concentration of D-glucose(30mM)for 48 hours.After modeling successfully,lmg/ml ginsenoside Rgl was added to culture for 48 hours.Then,Collection of cells in each group.Detection of Vimentin protein expression in cells by immunofluorescence staining and the proteins and RNAs of TGF-beta,Smad2,Smad3 and other related fibrosis factors sucn as alpha-SMA,N-cadherin,Vimentin,ZEB 1,MMP-2,MMP-9 and TIMP-2 were detected by Western blotting and RT-PCRResults:(1)Western blotting and RT-PCR results showed that:compared with normal control group,high glucose induces interstitial transformation of HRMECs cells,and up-regulation the expression of TGF-β,Smad2,Smad3 and fibrosis related factors alpha-SMA,N-cadherin,Vimentin,ZEB 1,MMP-2 and MMP-9,down-regulation the expression of Timp-2,the difference was statistically significant(P<0.05).Rgl intervention group cells compared with the high glucose group cells,the expression of TGF-β,Smad2,Smad3 and fibrosis related factors alpha-SMA,N-cadherin,Vimentin,ZEB 1,MMP-2 and MMP-9 were down regulated and TIMP-2 was up regulated,the difference between the two groups was statistically significant(P<0.05).(2)Immunofluorescence staining results:compared with the normal control group,the expression of Vimentin in the HRMECs cells of high glucose group was significantly increased,and the protein staining was significantly enhanced.After the addition of Rgl,the expression was obviously weakened.Conclusions:(1)The TGF beta/Smart signaling pathway is involved in the development of diabetic retinopathy.High glucose promotes retinal mesenchymal transition and induces retinal fibrosis.(2)ginsenoside Rgl has anti-fibrosis effect.(3)In the process of diabetic retinopathy,ginsenoside Rg1 reduces the degree of retinal fibrosis by inhibiting the TGF beta/Ssad signaling pathway. |
