Mesenchymal Stem Cells Derived Exosomes Protect Müller Cells From High Glucose-induced Cell Damage

Part 1Objective:In the current study,we evaluated the protective effects of Mesenchymal stem cells derived exosomes on high glucose-induced injury in Muller cells and then analyzed its mechanism.Methods:The cultured Muller cells were divided into normal control group(5mmol/L glucose),high glucose group(50 mmol/glucose),MSCs-exo treated group(50 mmol/L glucose+25 μg/mL MSCs-exo).The cell viability was detected by CCK8 assay,and cell apoptosis was detected by Annexin V-FITC apoptosis assay kit.The mRNA expression of protein TXNIP,NLRP3,ASC and CASP1 were detected by RT-PCR assay.Results:Compared with the normal control group,the cell viability of the high glucose group was decreased(P<0.01).The apoptosis rate of the group was higher than that in the normal control group(P<0.01).The mRNA expression of TXNIP,NLRP3,ASC,and CASP1 in the high glucose group was higher than in the normal control group(P<0.05).The result was statistically significant.Compared with the high glucose group,the cell viability of MSCs-exo treated group was significantly increased(P<0.01),and the apoptosis rate of MSCs-exo treated group was significantly decreased(P<0.01).The mRNA expression of genes TXNIP,NLRP3,ASC,and CASP1 in the MSCs-exo treated group was decreased(P<0.01).The result was statistically significant.Conclusion:High glucose can induce Müller cells apoptosis by activating TXNIP/NLRP3 inflammasomes axis.MSCs-exo attenuates apoptosis in Müller cells stimulated with high glucose via inhibiting TXNIP/NLRP3 inflammasomes axis.Part 2Objective:We aimed to explore the effect of knocking out TXNIP on high glucose-induced Müller cells.Methods:The cultured Müller cells were divided into normal control group(5mmol/L glucose),high glucose group(50 mmol/glucose),si-TXNIP treated group(50 mmol/L glucose+si-TXNIP),si-TXNIP-NC treated group(50 mmol/L glucose+si-TXNIP-NC).The cell viability was detected by CCK8 assay,and cell apoptosis was detected by Annexin V-FITC apoptosis assay kit.The mRNA expression of protein TXNIP,NLRP3,ASC,and CASP1 was detected by RT-PCR assay.Results:Compared with the high glucose group and si-TXNIP-NC treated group,the cell viability of si-TXNIP treated group was significantly increased(P<0.01),and the apoptosis rate of si-TXNIP treated group was significantly decreased(P<0.01).The mRNA expression of protein TXNIP,NLRP3,ASC,and CASP1 in the si-TXNIP treated group was decreased(P<0.05).The result was statistically significant.Conclusion:Knocking out TXNIP-NLRP3 inflammasome axis protects Muller cells from high glucose-induced cell damage.Part 3Objective:In this part,we aimed to explore the effective components of MSCs-exo that reduced cell apoptosis caused by high glucose.Methods:The cultured Müller cells were divided into normal control group(5mmol/L glucose),high glucose group(50 mmol/glucose),miR-93-5p-mimic treated group(50 mmol/L glucose+miR-93-5p-mimic),miR-93-5p-mimic-NC treated group(50 mmol/L glucose+miR-93-5p-mimic-NC).The cell viability was detected by CCK8 assay,and apoptosis was detected by Annexin V-FITC apoptosis assay kit.Results:Compared with the high glucose group and miR-93-5p-mimic-NC treated group,the cell viability of the miR-93-5p-mimic treated group was significantly increased(P<0.01),and the apoptosis rate of the miR-93-5p-mimic treated group was significantly decreased(P<0.01).The result was statistically significant.Conclusion:MSCs-exo contains numerous miRNAs targeted TXNIP-NLRP3 inflammasome.miR-93-5p is the most effective one that reduces cell apoptosis caused by high glucose.