Effect Of Acupuncture On Prolonging The Thrombolytic Time Window Of Cerebral Infarction And Regulating Neuron's Caspase-independent Apoptosis Pathway

Objective:To observe the effect of acupuncture(Acupunct)on cerebral infarction volume and expression of poly ADP ribose polymerase-1(PARP1),apoptosis-inducing factor(AIF)and endonuclease G(Endo-G)in the cerebral cortex tissue at different time-points after cerebral ischemia(CI)in acute cerebral infarction rats,so as to explore its underlying mechanisms in prolonging time window of thrombolysis.Methods:SPF level rats were randomly divided into sham operation,model,intravenous thrombolysis(IVT)-4.5h,IVT-6h,IVT-9h,Acupunct+IVT-4.5h,Acupunct+IVT-6h,Acupunct+IVT-9h groups(n=6 in each group).The CI model was established by using modified autologous thromboembolism via the right common carotid artery.Two hours after modeling,rats of the Acupunct groups received Acupunct stimulation of bilateral "Neiguan"(PC6)and Renzhong(GV26)for 30 min.Throm-bolysis was conducted by injection of Recombinant human tissue-type plasminogen activator(rt-PA,1 Omg/kg)via caudal vein.The neurological deficit was assessed with reference to Longa's and other researchers'methods(0—7points).Triphenyltetrazolium chloride(TTC)staining was used to assess the cerebral infarction volume,and the expression of cerebral PARP1,AIF and Endo-G proteins detected by Western blotting.Results:(1)Rats neurobehavioral scores:Contrasted with sham operation group,the neurobehavioral scores of other groups were obviously higher which means molding succeed and statistical differences existed(P<0.01).Compared with the modeling group,neurological scores were significantly reservedin the IVT-4.5 h,Acupunct+IVT-4.5h and Acupunct+IVT 6h groups(P<0.05).Nerve functions were all improved and enhanced after acupuncture intervention within 6h;The neurobehavioral scores between Acupunct+IVT 9h groupsand IVT-9hgroups are almost the same,statistical differences were not existed(P>0.05).(2)Percentage of cerebral infarction volume in each group:Compared to the sham operation group,the cerebral infarction volume of other groups were significantly increased which means molding succeed and statistical differences existed(P<0.01);Compared with the modeling group,percentages of cerebral infarction volume were significantly reservedin the IVT-4.5h,Acupunct+IVT-4.5h and Acupunct+IVT 6h groups(P<0.05).Nerve functions were all improved and enhanced after acupuncture intervention within 6h;The percentages of cerebral infarction volume between Acupunct+IVT 9h groups and IVT-9h groups are almost the same,statistical differences were not existed(P>0.05).(3)Comparison of the expression of PARP1,AIF and Endo-G mRNA in each group:Compared with the sham operation group,the expression volumes of PARP1,AIF and Endo-G mRNA were all increased significantly in other groups.Statistical differences were existed(P<0.01,P<0.05).Compared with the modeling group,expression volume of PARP1,AIF and Endo-G mRNA were significantly reservedin the IVT-4.5h(P<0.05).In addition,the expression volume decreased of PARP1,AIF and Endo-G mRNA after acupuncture intervention;Compared with,the modeling group except time window,expression volume of PARP1,AIF and Endo-G mRNA were significantly reservedin the Acupunct+IVT 6h(P<0.05);Compared with the modeling group,expression volume of AIF was significantly decreasedin the IVT6h(P<0.05);Acupuncture intervention can dramatically decrease the expression volume of PARP1 and AIF besides the 6h time window.Statistical differences were existed(P<0.05).Compared with the modeling group,expression volume of PARP1,AIF and Endo-G mRNA were not significantly reservedin the IVT9h and Acupunct+IVT 9h(P>0.05),statistical differences were not existed(P>0.05).(4)Comparison of the cortical protein expression of PARP1,AIF and Endo-G in each group:Compared with the sham operation group,the cortical protein expression volumes of PARP1,AIF and Endo-G were all increased significantly in other groups.Statistical differences were existed(P<0.01,P<0.05).Compared with the modeling group,the cortical protein expression volume of PARP1 was significantly reservedin the IVT-4.5h and Acupunct+IVT 4.5h(P<0.05).The cortical protein expression volume of AIF and Endo-G were significantly reservedin the IVT-4.5h,acupunct-t-IVT4.5h,and acupunct+IVT6h(P<0.05).Acupuncture intervention can dramatically decrease the cortical protein expression of PARP1,AIF and Endo-G in the 4.5h time window.Statistical differences were existed(P<0.05).Compared with IVT6h,the cortical protein expression of AIF dramatically decreased in the acupunct+IVT6h(P<0.05).Compared with IVT9,the cortical protein expression of AIF dramatically decreased in the acupunct+IVT9(P<0.05).Conclusion:?Perfect effects may achieved when cerebral infarction was treated within 4.5h time window by thrombolytic therapy.?When acupuncture was intervened timely,extended time window for thrombolysis of cerebral infarction would occur.This experimental study shows that acupuncture can lengthen the safety time window to 6h.?Acupuncture intervention can reduce the expression of mRNA and protein in PARP1,AIF and Endo-Gapoptosis related factors,which means acupuncture can achieve extended time window for thrombolysis of cerebral infarction through Caspase nondependent apoptotic pathways modification.