Study On The Role Of Lithium Chloride In The Wound Healing Model Of Diabetic Corneal Epithelial Lesions In Vitro

Background Diabetes is one of the major health problems of global concern.Late-stage diabetes can cause systemic complications,and diabetic corneal epithelial disease is one of the common ocular complications,which seriously affects the vision of patients and even causes blindness.Objective The purpose of this study is to use human corneal epithelial cells(HCEC)cultured in a high-glucose environment as the research object,through in vitro experiments to study the role of lithium chloride in the in vitro diabetic corneal epithelial defect healing model,and explore its mechanism of action,in order to provide more theoretical basis for the diagnosis and treatment of diabetic corneal epithelial disease.Methods Culture HCEC in vitro.Experimental groups: normal group(N group),normal+ lithium chloride group(N+LICL group),mannitol group(Ma group),mannitol + lithium chloride group(Ma+LICL group),high glucose group(G Group)and high sugar + lithium chloride group(G+LICL group).(1)Detect cell proliferation in each group by cell count and CCK-8 experiment;(2)Detect cell migration in each group by cell scratch test;(3)Detect cell apoptosis in each group by cell apoptosis kit;(4)Detect the gene level expression of Wnt/β-catenin signaling pathway related factors Wnt3,7a,β-catenin,Cyclin D1 in each group and the cells after the scratch treatment established cell injury model by q-PCR method.Results Through experimental research on HCEC cells,(1)Cell scratch experiment showed that the migration rate of HCEC cells in the N group and Ma group was significantly higher than that of the G group(P<0.05),and the HCEC cell migration rate of the G+LICL group was significantly higher than that of the G group.(P<0.01),the migration rate of HCEC cells in the G+LICL group was significantly higher than that in the N group and Ma group(P<0.05);(2)CCK-8 cell proliferation experiments showed that at 6 hours: HCEC cells in the G+LICL group The survival rate of HCEC was significantly higher than that of G group(P <0.05);at 12 hours: the survival rate of HCEC cells in G group was significantly lower than that of N group(P<0.01)and Ma group(P<0.05),G+LICL group HCEC The survival rate of the cells was significantly higher than that of the G group(P <0.05);at 24hours: compared with the N and Ma groups,the survival rate of HCEC cells in the G group was significantly reduced(P <0.05),and the HCEC cells in the G+LICL group Compared with the G group,the survival rate was significantly increased(P <0.01);(3)The apoptosis of each group was detected by the cell apoptosis kit by flow cytometry.It was shown that compared with the N group and the Ma group,the G group HCEC The apoptotic rate of cells increased significantly(P <0.01),and the apoptotic rate of HCEC cells in the G+LICL group was significantly reduced compared with that in the G group(P <0.01);(4)q-PCR results showed that: at 24 hours,Compared with group N,the expressions of β-catenin,Cyclin D1,Wnt7 a,and Wnt3 in the N+LICL group were significantly increased(P<0.01);compared with the G group,β-catenin(P<0.05)and Cyclin in the G+LICL group The expression of D1(P<0.01)was significantly increased;the expression of β-catenin was significantly lower in group G compared with group N(P<0.01);the high glucose + lithium chloride scratch group(GH+LICL group)was compared with high glucose Compared with the scratch group(GH group),the expressions of β-catenin,Cyclin D1,and Wnt7 a were significantly higher(P<0.05).Conclusion 125μM exogenous lithium chloride can promote the migration and proliferation of HCEC cells under high glucose culture,inhibit HCEC cell apoptosis under high glucose culture,and promote damage and repair of HCEC cells under high glucose culture,which may be activated by this effect Wnt/β-catenin signaling pathway is related.