基于Raf激酶抑制剂蛋白探讨针刺对脑出血大鼠神经保护的作用机制OA

Objective To explore the mechanism of acupuncture on neuroprotection in rats with cerebral hemorrhage based on Raf kinase inhibitor protein(RKIP).Methods Fifty healthy,clean-grade male Sprague-Dawley(SD)rats were divided into a sham operation group,a cerebral hemorrhage group,an acupuncture+empty vector group,an acupuncture group,or an acupuncture+RKIP group using the random number table method,with 10 rats in each group.Except for the sham operation group,cerebral hemorrhage rat models were established in the other groups.The sham operation group and the cerebral hemorrhage group received placebo acupuncture(the bases were only fixed onto the head acupoints,without skin penetration),while the acupuncture+empty vector group,the acupuncture group,and the acupuncture+RKIP group received acupuncture treatment.After 10 consecutive days of intervention,neurological behavior scores,pole climbing test scores,and brain tissue water content were compared among the groups.Changes in brain tissue were observed via hematoxylin and eosin(HE)staining.Neuronal apoptosis rate was calculated with terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling(TUNEL)staining.Furthermore,TNF receptor-associated factor 6(TRAF6)and phosphorylated NF-κB p65(p-p65)expression levels were measured by Western blot.Results Compared with the sham operation group,the cerebral hemorrhage group exhibited increased neurological behavior scores,brain tissue water content,and neuronal apoptosis rate,along with up-regulated TRAF6 and p-p65 expression levels and decreased pole climbing test scores(P<0.05).Compared with the cerebral hemorrhage group,the acupuncture+empty vector group showed decreased neurological behavior scores,brain tissue water content,and neuronal apoptosis rate,along with down-regulated TRAF6 and p-p65 expression levels and increased pole climbing test scores(P<0.05).Compared with the acupuncture+empty vector group,the acupuncture group displayed decreased neurological behavior scores,brain tissue water content,and neuronal apoptosis rate,along with down-regulated TRAF6 and p-p65 expression levels and increased pole climbing test scores(P<0.05).Compared with the acupuncture group,the acupuncture+RKIP group demonstrated decreased neurological behavior scores,brain tissue water content,and neuronal apoptosis rate,along with down-regulated TRAF6 and p-p65 expression levels and increased pole climbing test scores(P<0.05).According to HE staining results,the sham operation group exhibited normal brain tissue structure,with neatly arranged cells.The cerebral hemorrhage group showed brain tissue cell necrosis,with karyopyknosis and disordered cell arrangement.The acupuncture+empty vector group displayed decreased necrotic cells,with irregular cell arrangement.The acupuncture group and the acupuncture+RKIP group demonstrated gradually decreased necrotic cells,with orderly cell arrangement.Conclusion For rats with cerebral hemorrhage,acupuncture combined with RKIP protects neurological function,which is related to the TRAF6/NF-κB signaling pathway inhibition.