Effect Of Electroacupuncture On Caspase-3 Protein Expression In Ischemic Cortex Of Rats With Hyperlipemia Combined With Cerebral Ischemia

Objective:To observe the effect of electroacupuncture(EA)stimulation of Fenglong(ST40),Sanyinjiao(SP6)plus manual acupuncture(MA)stimulation of Suigou(GV26)and Baihui(GV20)on Caspase-3 protein expression in the cerebral cortex of rats with hyperlipemia and cerebral ischemia(HL-CI),so as to reveal its mechanisms underlying improvement of HL-CI.Methods:One hundred and five rats were randomly divided into normal control,sham operation,model,EA group I(EA+MA was given for7 and14 days,i.e.7 days before CI,and 7 days more after CI)and EA group II(EA+MA was given for only1 and 7 days after HL-CI),with 7 rats being in each group.The HL-CI model was established by feeding the animals with high fat forage for 6 weeks and then making an occlusion of the unilateral middle cerebral artery by regional application of quantitative paper adsorbing 50%FeC13 solution(10 ?L).Rats of the sham operation group were treated with the same procedures only without application of FeC13 solution.For rats of the EA group I,EA(1-3 mA,15Hz)was applied to bilateral acupoints SP6 and ST40(for 20 min),and MA stimulation applied to bilateral GV26 and GV20,which was conducted once daily for 7 days after 6 weeks' high fat forage feeding,followed by 1?7 days' EA+MA intervention again after CI modeling.For rats in the EA group?,EA+MA was applied to the same 4 acupoints once a day for 1?7 days only after CI modeling.The neurological impairment was assessed by Zea Longa' s scoring.The blood sample was taken from the abdominal aorta for measuring the contents of serum cholesterol(CHO),triglyceride(TG)and high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C).Cerebral infarction volume were measured by TTC staining.Pathological changes of the cerebral cortex were observed after H.E.staining,and the expression of Caspase-3 was analyzed by immunohistochemistry.The pathological changes of ischemic cerebral cortex tissue were displayed by H.E.staining.Results:1.Effects of electro acupuncture on neurobehavioral function in rats:Compared with the normal group,sham operation group,model group,EA group and I EA group II neurological function score after the model 1,3 and 7 days were significantly different(P<0.05);compared with the model group,EA group ??? after modeling,1,3,7 days were significantly different(P<0.05);compared with EA group ?,1,3 days after modeling,electroacupuncture group I score decreased significantly,there was significant difference(P<0.05),7 days after modeling,no significant difference between the EA I group and II group(P>0.05);compared with EA II group,electroacupuncture group ? 1 days after modeling,significant difference in CHO,HDL-C and LDL-C content were(P<0.05),there was no significant difference in TG content(P>0.05),7 days after modeling,TG,significant difference in CHO,HDL-C and LDL-C content were(P<0.05)I;two EA group,CHO,HDL-C and LDL-C content were significantly different(P<0.05),there was no significant difference in TG content(P>0.05);two EA II group,significant difference in CHO,TG and LDL-C content were(P<0.05),there was no significant difference in HDL-C content(P>0.05).2.Effects of EA on blood lipids(cholesterol,triglyceride,high density lipoprotein and low density lipoprotein)in ratsBlood lipids showed that compared with the normal group,sham operation group,model group rats after 1 and 7 days,significant difference in CHO,TG,HDL-C and LDL-C content were(P<0.05);compared with the model group,EA group I 1,7 days after modeling,CHO,TQ HDL-C and the content of LDL-C had significant difference(P<0.05),electroacupuncture group II model after 1 days,there was significant difference between the content of TG(P<0.05),CHO,HDL-C and LDL-C were no significant difference(P>0.05),7 days after modeling,TG,significant difference in CHO,HDL-C and LDL-C content have(P<0.05);3.Effect of EA on liver histomorphology in ratsThe results of HE staining showed that after 1 days,the liver cells of normal group and sham operation group had no degeneration and necrosis,the cells were arranged in order and the structure was clear.The model group after modeling for 1 days,visible lymphocyte aggregation,liver plate cord like structure was destroyed,hepatic sinusoids are smaller and less rare punctate necrosis;7 days after modeling,liver cell swelling significantly,liver sinus structure is not clear,visible steatosis,ballooning degeneration is typical disease widely.Physical performance is more obvious than model group of 1 days of serious.After 1 and 7 days,the pathological changes of hepatic cells were lighter in I group and II group,and the shape of liver tissue,hepatic sinusoid,hepatic lobule structure was not clear,inflammatory cells were significantly lower than that of the model group.4.Effect of EA on cerebral infarction volume in ratsThe results of TTC staining showed that 1 days after cerebral ischemia,the model of cerebral ischemia showed typical ischemic changes.Compared with the sham operation group,model group,EA group ??,infarct volume accounted for significant differences(P<0.05);compared with the model group,I EA group the infarct volume accounted for significant differences(P<0.05),electroacupuncture group ? infarct volume accounted for no significant difference(P>0.05);compared with EA ? group,electroacupuncture group I infarct volume ratio decreased,there was significant difference(P<0.05).5.Effect of EA on brain morphology in ratsThe results of HE staining showed that the nexurons in normal group and sham operation group were normal,arranged in order and the nucleolus was clear.The model group after modeling for 1 days,showed mild gliosis,neuronal swelling,7 days after modeling visible interstitial cell edema,the death of a large number of neurons showed cribriform necrosis,necrotic tissue peripheral inflammatory cells infiltration;tissue hippocampus CA1 staining showed that the number of normal group and sham operation group in hippocampus CA1 compact nucleus,nucleus morphology were normal.In the model group,1 days after the operation,there was no softening foci,and hippocampal cells shrank less.The number of neurons in the hippocampus was decreased,and the cell nucleus was reduced and the levels were disordered after 7 days.The damage of nerve cells in EA group I and group II was significantly lower than that of model group,the number of surviving neurons increased,the hippocampal structure was improved,and the arrangement was more orderly.6.Effect of EA on Caspase-3 in the cortex of ischemic foci in rats with hyperlipidemia and cerebral ischemiaThe expression of Caspase-3 protein in ischemic cortex of rats in normal group and sham operation group was weakly positive.Compared with the normal group,sham operation group after 1 and 7 days,the number of Caspase-3 positive cells and the average optical density values were not significantly different(P>0.05);compared with sham operation group,model group rats after 1 and 7 days,the number of positive cells in ischemic cortex and the average optical density value significantly significant differences(P<0.05);compared with the model group,EA group ? 1,7 days after modeling,were obviously different between the number of Caspase-3 positive cells and the average optical density values(P<0.05),no significant difference between the 1 days the number of Caspase-3 positive cells after modeling Electroacupuncture group ?(P>0.05).The average optical density had significant difference(P<0.05),electroacupuncture group ? model after 7 days,the number of Caspase-3 positive cells and the average optical density had significant difference(P<0.05);compared with EA II group,electroacupuncture group ? 1,7 days after modeling,the number of Caspase-3 positive cells The average optical density values were significantly different(P<0.05);two EA I group,the number of Caspase-3 positive cells and the average optical density had significant difference(P<0.05);two EA II group,the number of Caspase-3 positive cells and the average optical density had significant difference(P<0.05).Conclusion:1.Electro acupuncture can decrease the neurological function score,and early intervention of cerebral ischemia has obvious effect on improving neurological symptoms,the immediate effect of acupuncture on the improvement of neurological function.2.EA can reduce the content of CHO,TG and LDL-C in serum,increase the content of HDL-C,with the time of ischemia,the damage is more serious,and the effect of Electroacupuncture on HDL-C is not obvious.3.Cerebral ischemia can aggravate the liver injury induced by hyperlipidemia,and EA can improve the pathological changes of liver tissue.4.Electro acupuncture can improve the pathological changes of the brain tissue in ischemic area,reduce the degeneration and loss of neurons,nerve cell swelling and inflammatory cell infiltration.5.Early electroacupuncture intervention can reduce infarct volume and reduce brain injury in rats.6.The content of electroacupuncture can down regulate cortical Caspase-3 protein in focal ischemia in rats with hyperlipidemia combined with cerebral ischemia,and early acupuncture intervention lowered obviously,indicating its resistance to apoptosis,expression of Caspase-3 protein may be one of the mechanisms of acupuncture effect.