AIM:To evaluate the effectiveness of knock-down of heat shock protein 47(HSP47)on conjunctival bleb scarring in a rat model and its possible mechanism.METHODS:Male Sprague–Dawley rats were used for glaucoma filtratio...AIM:To evaluate the effectiveness of knock-down of heat shock protein 47(HSP47)on conjunctival bleb scarring in a rat model and its possible mechanism.METHODS:Male Sprague–Dawley rats were used for glaucoma filtration surgery(GFS)and were treated with either phosphate buffered solution,shControl,mitomycin C,or sh-HSP47 using a microsyringe immediately after GFS.The morphology of filtering blebs was observed postoperatively.The levels of HSP47 were analyzed at 2,5,8,and 11d after GFS via real‑time quantitative polymerase chain reaction(PCR)and Western blot.The silencing effect of HSP47,the expression of collagen I and III,and the potential signaling pathways of HSP47 during scarification were explored 11d post GFS.The protein levels of transforming growth factor-β1(TGF-β1),phospho-Smad2(pSmad2),phospho-Smad3(p-Smad3),and phospho-p38(p-p38)were also analyzed using Western blot.RESULTS:Sh-HSP47 treatment significantly prolonged the functional filtration bleb retention.The levels of HSP47 were increased significantly at 5,8,and 11d postoperatively compared to the control group(P<0.05,P<0.01,and P<0.001).The levels of HSP47 protein at day 11 postoperatively were significantly down-regulated after HSP47 silencing using sh-HSP47 adenovirus transfection(P<0.01).Expression levels of collagen I and III within the blebs were significantly reduced in the absence of HSP47(P<0.01).Moreover,the protein levels of TGF-β1,p-Smad2/3,and p-p38 were dramatically inhibited after treatment with sh-HSP47(P<0.01).CONCLUSION:The inhibitory effects of HSP47 knockdown on scarring after GFS have the potential to be an efficacious therapeutic option for the treatment of conjunctival bleb scarring.展开更多
Heat shock protein 47 (HSP47) is a collagen-specific molecular chaperone that is required for mo-lecular maturation of various types of collagens. Many studies have shown a close association be-tween increased express...Heat shock protein 47 (HSP47) is a collagen-specific molecular chaperone that is required for mo-lecular maturation of various types of collagens. Many studies have shown a close association be-tween increased expression of HSP47 and excessive accumulation of collagens in scar tissues of various human fibrotic diseases. However, the role of HSP47 in formation of scar after glaucoma filtration surgery is still unclear. In this study, we deleted the expression of HSP47 in human tent on fibroblasts (HTFs) by virus infection, and then the proliferation and collagen synthesis were compared between HSP47 deletion cells and control upon TGF-β1 stimulation. Our data showed that HSP47 deletion could significantly inhibit the proliferation and collagen synthesis of HTFs upon TGF-β1 stimulation, HSP47 gene suppression might be a novel method to against the formation of scar after glaucoma surgery.展开更多
基金Supported by the National Natural Science Foundation of China(No.81500719)Shaanxi Science and Technology Project(No.2022SF-434)Xi’an Science and Technology Project(No.21YXYJ0044).
文摘AIM:To evaluate the effectiveness of knock-down of heat shock protein 47(HSP47)on conjunctival bleb scarring in a rat model and its possible mechanism.METHODS:Male Sprague–Dawley rats were used for glaucoma filtration surgery(GFS)and were treated with either phosphate buffered solution,shControl,mitomycin C,or sh-HSP47 using a microsyringe immediately after GFS.The morphology of filtering blebs was observed postoperatively.The levels of HSP47 were analyzed at 2,5,8,and 11d after GFS via real‑time quantitative polymerase chain reaction(PCR)and Western blot.The silencing effect of HSP47,the expression of collagen I and III,and the potential signaling pathways of HSP47 during scarification were explored 11d post GFS.The protein levels of transforming growth factor-β1(TGF-β1),phospho-Smad2(pSmad2),phospho-Smad3(p-Smad3),and phospho-p38(p-p38)were also analyzed using Western blot.RESULTS:Sh-HSP47 treatment significantly prolonged the functional filtration bleb retention.The levels of HSP47 were increased significantly at 5,8,and 11d postoperatively compared to the control group(P<0.05,P<0.01,and P<0.001).The levels of HSP47 protein at day 11 postoperatively were significantly down-regulated after HSP47 silencing using sh-HSP47 adenovirus transfection(P<0.01).Expression levels of collagen I and III within the blebs were significantly reduced in the absence of HSP47(P<0.01).Moreover,the protein levels of TGF-β1,p-Smad2/3,and p-p38 were dramatically inhibited after treatment with sh-HSP47(P<0.01).CONCLUSION:The inhibitory effects of HSP47 knockdown on scarring after GFS have the potential to be an efficacious therapeutic option for the treatment of conjunctival bleb scarring.
文摘Heat shock protein 47 (HSP47) is a collagen-specific molecular chaperone that is required for mo-lecular maturation of various types of collagens. Many studies have shown a close association be-tween increased expression of HSP47 and excessive accumulation of collagens in scar tissues of various human fibrotic diseases. However, the role of HSP47 in formation of scar after glaucoma filtration surgery is still unclear. In this study, we deleted the expression of HSP47 in human tent on fibroblasts (HTFs) by virus infection, and then the proliferation and collagen synthesis were compared between HSP47 deletion cells and control upon TGF-β1 stimulation. Our data showed that HSP47 deletion could significantly inhibit the proliferation and collagen synthesis of HTFs upon TGF-β1 stimulation, HSP47 gene suppression might be a novel method to against the formation of scar after glaucoma surgery.
