The Regulation Of Extracellular Hsp90 In Promoting Posterior Capsule Opacification

Posterior capsule opacification(PCO),also known as posterior cystic turbidity,is the most common complication after cataract artificial crystal replacement.Postoperative PCO incidence is 97%in children under 12 years of age and 20-50%in adults.The main pathological features of PCO are the proliferation,migration,Epithelial-Mesenchymal Transition(EMT)and collagen deposition of residual epithelial cells after micro-environmental regulation,but the regulatory mechanism is not entirely understood.At present,there is no effective drugs for PCO therapy,and Nd:YAG laser cut after the cystic membrane is the only effective method,however,it is easy to cause complications such as short-term eye pressure rise,iritis,retinal tearing and separation,macular edema,corneal edema,artificial lens dislocation into the glass and artificial lens depression.Moreover,the Nd:YAG laser treatment does not affect the abnormal growth of residual crystalline epithelial cells and their changes in the microenvironment of the eye aqueous humor.Therefore,it is of great significance to study the regulation of microenvironment for the development of PCOFirst of all,we pay attention to the micro-environment molecules in the development of PCO.Because relative to the intracellular target,inhibit the activity of key extracellular molecules in the pathological process can not only block disease progression,but also reduce the impact on normal cellular physiological state.We found that the crystalline epithelial cells secrete heat shock protein 90(eHsp90)in earlier experiment.By reviewing the literatures,it is found that eHsp90 can promote tumor cell EMT,increase tumor cell migration and invasion ability,and in tissue damage repair process,eHsp90 can promote the healing of skin tissue.Therefore,we speculate that the microenvironment eHsp90 is involved in the development of PCOOur project takes rat lens capsular bag culture as a model,discusses the mechanism of the role of microenvironment eHsp90 in the development process of PCO,which hope to provide new perspectives for the treatment of PCOBuilding a PCO model of rat in vitro,collect cultured phenotypes and then use mass spectroscopy(MS)to assist in the analysis of possible secretion components in the supernatant,and finally use Western Blotting to verify whether the secretion of the upper clear contains eHsp90 protein.Western Blotting assay results show that in in vitro PCO models,crystalline epithelial cells are able to secrete eHsp90 protein to the medium.Building the New Zealand rabbit PCO model in vivo,observe and record the development of PCO,collect the PCO formation model of eye aqueous humor,and then use immunoprecipitation technology(IP)to detect the same model rabbit normal eye and PCO model eye aqueous humor eHsp90 protein content difference The results show that compared with the normal eye aqueous humor,the PCO eye aqueous humor contains more eHsp90 protein.Collect the PCO model at different incubation points and then detect the change of the content of eHsp90 protein,and use high-speed centrifugal and Proteinase K processing culture on the supernatant,study the secretion of eHsp90 in the PCO model.The crystalline epithelial cells in the PCO model will continuously secrete eHsp90 protein to the medium with the increase of culture time,and most of the secretion of this protein is secreted in free form.Using different concentrations of TGFβ2 and TGF beta inhibitor SB431542 to treat the human crystalline epithelial cell line SRA 01/04 and collect cultured top-clearing,Western Blotting verified the effects of TGF beta 2 of different treatment concentrations on eHsp90 secretion.TGFβ2 can promote the secretion of Hsp90 protein in human crystalline epithelial cells SRA 01/04,and has dose dependence,when using SB431542 to inhibit the activity of TGF β receptors can inhibit the secretion of Hsp90 protein.The PCO model was treated using TGFβ2 and the relevant inhibitors in the TGFβpathway,and then the development of PCO in different processing groups was observed and the content of eHsp90 protein in the supernatant was observed.TGF beta 2 can promote the development of PCO and the formation of the Lentoid bodies(LBs),and the process can be inhibited by some of the inhibitors of the TGFP pathway,while the inhibitors of the TGFβ pathway inhibit the secretion of eHsp90 protein induced by TGFβ2.Successful purified human recombinant Hsp90 alpha protein.Using purified human Hsp90 alpha protein and TGF beta 2 to jointly process PCO model,observe the development and change of PCO.Using purified human Hsp90 alpha protein can synergistical with TGFβ2 to promote the development of PCO.All in one,after cataract surgery,residual crystalline epithelial cells are able to secrete Hsp90 protein to the extracellular of the cell,and the content of the extracellular Hsp90 protein increases showing time dependent.TGF beta 2 can promote the secretion of eHsp90 protein in crystalline epithelial cells,a process that can be inhibited by some inhibitors of the TGFβ signaling pathway.Extracellular Hsp90 protein can synergistical with TGFP2 to promote the development of PCO.