温通活血乳膏通过调控内质网应激对高糖诱导雪旺细胞凋亡的影响OA

Objective It is to observe the effect of Wentong Huoxue Cream(WHC)on apoptosis of high-glucose-in-duced Schwann cells and on protein kinase R-like endoplasmic reticulum kinase(PERK)/eukaryotic translation initiation factor 2 α(EIF-2α)/activating transcription factor 4(ATF4)/C/EBP-homologous protein(CHOP),and explore its possi-ble mechanism of action.Methods Rat Schwann RSC96 cells were taken and cultured,and the effects of different concen-trations of Wengtong Huoxue Cream and 4-PBA on cell proliferation were assessed by CCK-8 assay to determine the optimal concentrations of Wengtong Huoxue Cream and 4-PBA for the subsequent experiments.4 groups were set up in the subse-quent experiment:the control group was cultured with low-glucose DMEM medium(containing 25 mmol/L glucose),the high glucose group was cultured with high-glucose DMEM medium(containing 100 mmol/L glucose),the high glucose+WHC group was cultured with high-glucose DMEM medium containing 8 mg/mL WHC,the high glucose group+4-PBA group was cultured with high-glucose DMEM medium containing20 mg/mL4-PBA,and the high glucose+4-PBA+WHC group was cultured with high-glucose DMEM medium containing 20 mg/mL 4-PBA and 8 mg/mL WHC.After 24 hours of culture,the protein expressions of PERK,p-PERK,EIF-2α,p-EIF-2α,ATF4,CHOP,Bcl-2 and Caspase-3 in the cells were detected by Western blot analysis,the mRNA expressions of PERK,EIF-2α,ATF4,CHOP,Bcl-2 and Caspase-3 in the cells were detected by RT-qPCR,the cell apoptosis was assessed by flow cytometry.Results Compared with the control group,the relative protein expressions of p-PERK/PERK,p-EIF-2α/EIF-2α,ATF4,CHOP and Caspase-3,as well as relative mRNA expressions of PERK,EIF-2α,ATF4,CHOP and Caspase-3 were significantly increased,while the relative protein and mRNA expressions of Bcl-2 were significantly decreased,and the apoptosis rate was significantly increased in the high glucose group(all P<0.05).Compared with the high glucose group,the relative protein expressions of p-PERK/PERK,p-EIF-2α/EIF-2α,ATF4,CHOP and Caspase-3,as well as relative mRNA expressions of PERK,EIF-2α,ATF4,CHOP and Caspase-3 were significantly decreased,while the relative protein and mRNA expressions of Bcl-2 were significantly increased,and the apoptosis rates were significantly decreased in the high glucose+WHC group,high glucose group+4-PBA group and high glucose+4-PBA+WHC group(all P<0.05).Compared with the high glucose+WHC group and high glucose group+4-PBA group,the relative protein expressions of p-PERK/PERK,p-EIF-2α/EIF-2α,ATF4,CHOP and Caspase-3,as well as relative mRNA expressions of PERK,EIF-2α,ATF4,CHOP and Caspase-3 were significantly decreased,while the relative protein and mRNA expressions of Bcl-2 were significantly increased,and the ap-optosis rate was significantly decreased in the high glucose+4-PBA+WHC group(all P<0.05).Conclusion Wentong Huoxue Cream can inhibit high glucose-induced Schwann cell apoptosis via regulating endoplasmic reticulum stress through PERK/EIF-2α/ATF4/CHOP pathway to down-regulate expression of Caspase-3 and up-regulate expression of Bcl-2.