桃红四物汤对大鼠肢体缺血-再灌注损伤中miR-675、CaMKII、TNF-α的表达影响及干预作用OA

Objective To investigate the intervention effect of Taohong Siwu Decoction(桃红四物汤,THSWD)on the expressions of miR-675,CaMKII,and TNF-α in a rat model of limb ischemia-reperfusion injury(LIRI).Methods Seventy-eight male SD rats were randomly assigned to four groups:the control,model,miR-675 inhibitor,and THSWD groups.Except for the control group,LIRI was induced in the rats of the other groups.After successful modeling,skeletal muscle and serum samples were collected at 0,2,4,and 8 hours for analysis.HE staining was used to observe morphological and pathological changes in skeletal muscle,and DAPI staining was used to assess cell apoptosis.The expression levels of miR-675 and CaMKII mRNA were detected by RT-qPCR.CaMKII protein expres-sion was measured by Western blot(WB),and serum TNF-α level was measured by ELISA.Results HE staining revealed the most severe skeletal muscle injury in the model group,while the injury was alleviated in the miR-675 inhibitor and THSWD groups.DAPI staining indicated that the model group had the highest number of apoptotic cells and the highest apoptosis rate,which was significantly different from the other groups(P<0.05).At each corresponding time point,RT-qPCR and WB analyses showed that the expressions of miR-675 and CaMKII mRNA,as well as CaMKII protein,were significantly higher in the model,miR-675 inhibitor,and THSWD groups than in the control group(P<0.05).However,the expressions in the miR-675 inhibitor and THSWD groups were significantly lower than those in the model group(P<0.05).ELISA revealed that serum TNF-α levels in all experimental groups were higher than those in the control group(P<0.05).Notably,the levels in the miR-675 inhibitor and THSWD groups were significantly lower than those in the model group(P<0.05).Conclusion THSWD alleviates LIRI in rat skeletal muscle.The mechanism may be associated with its regulation of miR-675 expression,thereby modulating the expression of CaMKII and TNF-α and other inflammatory factors.