Effect of Xingshen Tongdu Acupuncture on Autophagy in Rats with Cerebral Ischemia-Reperfusion Injury
Objective To investigate the effects of Xingshen Tongdu acupuncture on rats after cerebral ischemia and reperfusion based on autophagy.Methods Male SD rats were randomly divided into 3 groups:sham control(Sham),ischemia and reperfusion(IR),IR plus Xingshen Tongdu acupuncture(Acu).The model was induced by middle cerebral artery occlusion(MCAO)followed by reperfusion.The acupuncture intervention was started 5 days before modeling to the rats of the Acu group,and the acupoints"Baihui""Sishencong"and"Zusanli"were selected for treatment,once a day until 24 h after reperfusion.Rats in the other 2 groups were only grabbed without acupuncture.After 24 h of reperfusion,cerebral infarct areas of rats were evaluated by triphenyltetrazolium chloride(TTC)staining in all groups.Ultrastructural morphologies of autophagy were observed by transmission electron microscope(TEM).Levels of microtubule-associated protein light chain 3B(LC3),coiled coil moesin like BCL2 interacting protein(Beclin-1),autophagy related protein(Atg)12,protein kinase B(Akt)and p-Akt were detected by Western blot and immunofluorescent staining.Results Compared with the Sham group,cerebral infarct areas of MCA territory were increased(P<0.05),and TEM indicated that neuronal autophagy was significantly activated in the rats of the IR group.Moreover,Beclin-1,Atg12 expression levels,and the ratio of LC3Ⅱ/I were significantly up-regulated(P<0.05),while the p-Akt level was decreased(P<0.05)in the rats of the IR group.Compared with the IR group,the cerebral infarct area decreased(P<0.05),while TEM revealed a relatively normal neuronal structure with fewer autophagosomes in the rats of Acu group.Furthermore,the expression of Beclin-1 and Atg12 as well as LC3 Ⅱ/I ratio decreased obviously(P<0.05),while the expression of p-Akt protein was significantly up-regulated(P<0.05)in the rats of Acu group.Conclusion The protective effects of acupuncture on cerebral ischemia and reperfusion injury may be related to inhibiting autophagy.
Xingshen TongduAcupunctureCerebral ischemia and reperfusion injuryAutophagy