| Objective: To explore the mechanism of the modified recipe of removing wind and benefiting damage to regulate the EMT process of lens epithelial cells and influence the proliferation,differentiation and migration of cells.Taking patients after cataract surgery as observation objects,observe the effects of modified prescription of removing wind and benefiting damage on the proliferation,differentiation and migration of lens epithelial cells.The EMT model rats were established by anterior capsule scratch experiment,and the correlation of inflammation signal axis of TLR4-NF-κB-NLRP3 in this process was studied.Method:1.Clinical observation: 72 patients after cataract surgery were included,all of whom were divided into 2 groups.The traditional Chinese medicine group was treated with routine western medicine group plus modified Chinese medicine granules for removing wind and benefiting loss.The control group was only intervened by routine western medicine.Chinese medicine is taken orally for 2 courses,and western medicine is used according to the course of treatment.Two years after operation,the patients’ long-distance naked eye vision,posterior capsule opacification percentage,opacification grade and the number of posterior capsule incision were recorded.2.Experimental research:(1)SPF SD rats were randomly divided into normal group and model group,the normal group was not treated,and the model group rats underwent anterior lens capsule scratch surgery to establish the rat lens EMT model.1,3,7,14 days after operation,1/4 rats in each group were randomly killed,and their right eyeballs were harvested,and the overall pathological changes were observed by HE staining.(2)SPF SD rats were randomly divided into normal group,model control group,middle dose group and high dose group.Except for the normal group,the other three groups were operated for modeling.After operation,the normal and model control groups were given double distilled water daily,and the Chinese medicine treatment group was given corresponding doses of Chinese medicine(0.55g/kg,1.10g/kg),both of which were 2ml.On the 3rd,7th and 14 th day after operation,1/3 rats in each group were randomly killed,and their right eyeballs were removed.The whole pathological changes were observed by HE staining,the changes of IL-1β and TNF-αwere observed by Elisa,and the expressions of TLR4,NF-κB and NLRP3 proteins were observed by immunohistochemistry and WB method.Results:1.Compared with the turbidity grade and the control group,the degree of posterior capsule opacification in Chinese medicine treatment group has statistical significance.(P<0.05),there was no significant difference in visual acuity and the number of posterior capsulotomy.2.Compared with the normal rats,the rats in the operation group have different degrees of pathological changes,the severity of which is proportional to the pathological time.3.No pathological changes were found in the normal group at all time points,but pathological changes were found in different degrees in the operation group,which were the heaviest in the model control group at different time periods.The middle dose group of Chinese medicine improved,and the high dose group of Chinese medicine improved significantly.4.Compared with the normal group,the expression of IL-1β in the model control group was significantly different on the 7th and 14 th day(P<0.05).After the intervention,there was a statistically significant difference in expression between the7-day middle-dose group and the 14-day middle-high-dose group(P<0.05).5.Compared with the normal group,the difference of TNF-α expression in the model control group in three periods was statistically significant(P<0.05);After the intervention,there was a significant difference in expression between the high-dose group and the high-dose group within 14 days(P<0.05).6.Compared with the normal group,the expression of TLR4 in the model control group was significantly different in three periods(P<0.05);After the intervention,the difference between the middle-dose group and the high-dose group was statistically significant 14 days after the operation,3 days after the operation and 7 days after the operation(P<0.05).7.Compared with the normal group,the expression of NF-κB in the model control group was significantly different on the 7th and 14 th day(P<0.05).After the intervention,there was a significant difference between the 7-day high-dose group and the 14-day middle-dose group(P<0.05).8.Compared with the normal group,the expression of NLRP3 in the model control group was statistically significant(P<0.05).After the intervention,there was a significant difference in expression between the high-dose group and the high-dose group within 14 days(P<0.05).Conclusion:1.The modified recipe of removing wind and benefiting damage can reduce the turbid area and turbidity severity of posterior capsule.2.The scratch operation of rat lens anterior capsule can be used for the establishment of EMT model of lens epithelial cells and the follow-up study.3.The modified recipe of removing wind and benefiting damage can effectively inhibit the secretion of IL-1β and TNF-α,inhibit the expression of TLR4,NF-κB and NLRP3 proteins,interfere with the EMT process of lens epithelial cells,and effectively prevent the proliferation,differentiation and migration of LECs.However,the optimal therapeutic dose still needs to be further explored.4.The modified recipe of removing wind and benefiting damage can regulate the EMT process of lens epithelial cells by regulating the inflammatory signal axis of TLR4-NF-κB-NLRP3. |
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