牦牛热休克蛋白B11通过抗凋亡缓解热应激对肾细胞的体外损伤OA北大核心CSTPCD

Heat shock protein beta-11(HSPB11)is a member of the small heat shock protein family,which has the func-tions of anti-apoptosis,promoting cell survival,and maintaining cell morphology.Under heat stress,dysautophagy is one of the pathogenesis of acute kidney injury,incomplete repair of kidney injury,and chronic kidney disease.The basic autophagy of kidney cells is key to maintaining kidney homeostasis,structure,and function.To study the role of yak HSPB11 on heat-stressed MDBK cell activity and apoptosis.Cells were divided into control group,heat stress group,and treatment group.PCR amplification,bioinformatics analysis,prokaryotic expression vector construction,CCK-8 method,acridine orange staining method,fluorescent enzyme labeling assay,fluorescence quantitative PCR,and other methods were used.A 757 bp amplified fragment of yak HSPB11 was obtained.Phylogenetic tree showed that the simi-larity between yak HSPB11 and Bos taurus HSPB11 was 99％.pET-32a-HSPB11 recombinant plasmid was successfully constructed and expression yielded about 35 kDa of yak HSPB11.The yak HSPB11 concentration of 10.0 μg/mL was screened to be the optimal concentration as the treatment group.The results showed that the relative activity of cells was significantly different between the control group and the heat stress group(P=0.004),and the relative activity of cells was significantly different between the heat stress group and the treatment group(P＜0.001).In the heat stress group,the cells contracted and the nucleus lysed in fragments,while in the treatment group,cell lysed was rare.In addition,the nucleic acid fluorescence intensity of MDBK cells in the heat stress group was significantly higher than that in the con-trol group and treatment group(P＜0.001).The mRNA expressions of caspase-3,caspase-9 and Bcl-2 in the heat stress group were significantly higher than those in the control group and treatment group(P＜0.001).In summary,it is hy-pothesized that yak HSPB11 alleviates MDBK cell damage by heat stress through the apoptotic pathway,which provides basic information for exploring the function of yak HSPB11.