| Objective:Bone marrow mesenchymal stem cells(BMSCs)were pretreated with atorvastatin(ATV)and injected into the vitreous cavity of diabetic rats.Observe whether the pretreated BMSCs can improve the retinal microangiopathy in diabetic rats,and investigate the therapeutic mechanism of BMSCs in diabetic retinopathy(DR).Methods:1.BMSCs were isolated and cultured,and their immunophenotypes were identified.2.BMSCs were treated with different concentration of ATV The expression of CXCR4 gene in BMSCs was detected by real-time quantitative polymerase chain reaction(RT-PCR)and Western blotting(WB);The proliferation ability of BMSCs after ATV pretreatment was detected by MTS cell proliferation assay.The migration ability of BMSCs after ATV pretreatment was detected by transwell chamber method and induced by SDF-1.The apoptotic degree of ATV pretreated BMSCs was detected by flow cytometry.3.The diabetic model was established by intraperitoneal injection of streptozocin STZ into SD rats.Four weeks later,BMSCs(5ul,3 × 104cells)pretreated with ATV were injected into the vitreous cavity of diabetic rats,blood glucose and body weight were measured 3 weeks before and 3 weeks after vitreous injection.The experimental animals were divided into four groups:normal control group(normal group),diabetic rats control group(Blank group),unpretreated BMSCs transplantation group(BMSC group),BMSCs treated with ATV transplantation group(ATV-BMSC group).Intervention treatment was carried out after the successful establishment of the model.Group MSC:untreated MSCs was injected into the vitreous cavity.Group ATV-MSC:the vitreous cavity was injected with ATV treated MSCs,In the normal control group and the Blank group,no special treatment was done.The retinal paraff-in sections in each group were detected by Western blotting and immunohistochemical method 1 week 2 weeks and 3 weeks after intravitreal injection.The changes of retinal pathological tissue,the expression of rhodopsin,NSE and TNF-? in the retina of rats in each group were observed,and the results were compared with each other.Results:1.It is susessful for isolation,extraction,culture and identification of SD rats BMSCs.2.After BMSCs was pretreated with ATV,the increased expression of CXCR4 gene was detected by Western blotting and RT-PCR.The proliferation ability of BMSCs cells in each group was not statistically significant(P>0.05)by MTS cell proliferation assay.Compared with other groups,the migration ability of ATV-MSCs was improved by transwell chamber method,and the difference was significant(p<0.05).The apoptotic degree of BMSCs was detected by flow cytometry,and the statistical difference was found between the two groups(p>0.05).3.Retinal paraff-in section HE staining showed that the degree of retinal inflammation in ATV-BMSCs group was less than that in other groups.Western-blotting and immunohistochemistry showed that the expression of Rhodopsin,NSE protein in the retina of ATV-BMSCs group was significantly higher than that in other groups(P<0.05),and the expression of IL-6 TNF-? protein was lower than that of other groups(P<0.05).Conclusion:1.In this study,BMSCs pretreated with ATV were found to be able to significantly increase migration and homing ability by increasing the expression of CXCR4 in vitro after ATV treatment,which provided a theoretical basis for the special treatment of MSCs for DR2.In this study,intravitreal injection of BMSCs pretreated with ATV was performed in diabetic rats for the first time.It was found that BMSCs pretreated with ATV could reduce the degree of retinal inflammation more effectively and regulate the express-ion of related factors.It can protect the retina in DRand open up a new idea for the treatment of DR by MSCs. |
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