Effect Of QingGuangAn Slow-release Agent To The TGF-β/Smad Signal Transduction Pathway Of The Fltering Scarring After Glaucoma Filtering Surgery

Object:To observe the effect of QingGuangAn slow-release agent to the TGF-β/Smad signal transduction pathway of the filtering scarring after glaucoma filtering surgery.To study the mechanisms of QingGuangAn slow-release agent,and preliminarily determine their safety.Method:Used high throughput drug screening which systempreliminary studied,extract 4 kinds of effective component from QingGuangAn.And 4 kinds of QingGuangAn effective components made into sustained-release film.48 healthy adult New Zealand rabbits were randomly divided into 8 groups,6 rabbits each group.A group:blank group,B group:model group,C group:MMC control group,Group D:effective constituents in 1 groups,Group E:effective constituents in 2groups,Group F:effective constituents in 3groups,Group G:effective constituents in 4groups,H group:the placebo group.The rabbits of A group without any treatment,all eyes of group B to H rabbits were underwent routine trabeculectomy and iridotomy.In group C,combined with MMC in operation.Corresponding QingGuangAn release membrane were implanted under conjunctvia of eyes from group D to G.Group H received placebo release membrane in conjunctiva.All rabbits were measured intraocular pressure in postoperative,to observe the shape of filtering bleb,checked the wound healing and general case of eyes.After 4 weeks,all rabbits were sacrificed.Clipped the scleral tissue of filtering from the operated eyes.To observe the histomorphology of filtering after HE staining.To detect the expression of mRNA and protein of TGF-β2、Smad3、Smad4 and collagen I in the scleral tissue of filtering by Real-time PCR and Western-blot,and analyzed statistically.Results:The intraocular pressure of group C and E rise slowly after operation.Compared with group A,B,D,F,G and H,there was significant difference between those group after 4 weeks(P<0.05).The filtering bleb of group C and E can maintain effective bleb after operation.The filtering bleb of group B was no functional or disappear after 2 weeks.4 weeks later,there were a few functional filtering bleb in group D,E,G and H.The outline of filtering bleb in group C and E was clear,and there were few fiber cells and inflammatory cells by HE staining.The outline of filtering blebs in other operation groups disappeared and were filled by a large number of fibroblasts and inflammatory cells.There was no statistic difference between operated groups in the expression of mRNA and protein of TGF-β2 afrter 4 weeks(P>0.05).There was significant difference between group C and other operated groups in the expression of mRNA and protein of Smad3 and Smad4 afrter 4 weeks(P<0.05),no statistic difference between group B,D,E,F,G and H(P>0.05).There was significant difference between group C,E and other operated groups in the expression of mRNA and protein of collagen I afrter 4 weeks(P<0.05),no statistic difference between group C and E(P>0.05).Conclusions:(1)To verify that the important reasons for the failure of filtering operation is that filtering bleb scarring to lead filter block,intraocular pressure raise,bleb lose function.(2)MMC and QingGuangAn effective constituents in 1 can effect the TGF-β/Smad signal transduction pathway of the filtering scarring after glaucoma filtering surgery.They can inhibit fibroblast to proliferate,type I collagen to express,and to reduce the scar tissue hyperplasia.They can obviously inhibit filtering bleb to form scar after glaucoma surgery.(3)QingGuangAn effective constituents in 2 and TGF-β can combine to TGF-β recepto competitively,so it can antagonize the effect of TGF-β.It can weak the function of TGF-β/Smad cell signal transduction pathways and reduce fibroblasts to secrete type I collagen.(4)QingGuangAn effective constituents in 2 can effect TGF-β/Smad cell signal transduction pathways directly.Beside it may affect inflammatory factor produce,reduce the effect of inflammatory factor to the filtering bleb,and reduce TGF-β indirectly.Thus it can effect TGF-β/Smad cell signal transduction pathways.(5)We have observed that the side effect of QingGuangAn effective constituents in 2 is small and more safe than MMC.It is expected to become a new type of safe drug that it can suppress filtering scar after glaucoma surgery,maintain low intraocular pressure and improve the success rate of operation。...