| Objective:By observing effects of electroacupuncture stimulation on two acupoints-"chize, Hegu" and "Zusanli, Sanyinjiao" on neural function deficit score, cerebral tissue morphology, cell apoptosis and p38MAPK phosphorylation ofcerebral ischemia reperfusion rats, we want to explore the possible mechanism that how electroacupuncture works against cerebral ischemia reperfusion injury and provide some experimental basis for clinical application of acupuncture on treatment and prevention of cerebral ischemia reperfusion.Method:250healthy male SD rats were randomly divided into5groups:sham operation group, model group, electro acupuncture group, blocker group and EA+blocker group,50rats/group. And then the rats were randomly divided into5sub groups:2h group,6h group, ID group,3D group and1W group accor-ding to the time point of reperfusion andevery sub group includes10rats. Except for the sham operation group, the cerebral ischemia/reperfu-sion model was established by modified thread occlusion methodin the other groups.The electroacupuncture group received electroacupuncture treat-ment once a day, the blocker group was in jectedp38MAPK blocker SB203580in corresponding brain areas30min before model establishment. EA+blocker group was injected SB203580before model establishment and then received electroacupuncture treatment. The nerve function defect score test sympt-oms of nerve defects in rats, to observe the morphological changes of brain tissue of rats with HE and TTC staining method, the detection of rat neural cell apoptosis index, TUNEL method, immunohistochemistry was used to dete-ct the expression of p-p38MAPK protein in ischemic area. Results:1.Comparison of neural function defect score：after modeling, compared with the sham operation group, score the remaining four rat nerve defect was significantly higher than that in sham operation group, there was a statistically significant difference (P<0.01), indicating the brain ische-mia reperfusion model success. With the reperfusion time, nerve deficit score decreased. After treatment, compared with model group, electroacupun-cture group, blocker group and EA+blocker group scores were lower than that of model group, and each group of3d and1W time point score decreased with statistical significance(p<0.05), group SB+EA1W time point score decreas-ed, there was a statistically significant (p<0.01). Comparison between elec-troacupuncture stimulation, blockers and combina-tion of three, no sta-tistical significance(p>0.05).2.To observe the morphology of brain cells of ischemic region of HE staining:sham operation group cell morphology, structure clear karyotheca integrity, clear nucleolus;model group cell gap widened, the neuron number, arranged in disorder, the nuclear membrane and the surrounding boundary is not clear,nucleolus pyknosis;Electroacupuncture group,blocking agent group and EA+blocker group of nerve tissue damage was lighter than in the model group, the cells were arranged in order, structure is complete, the cell body of mild to moderate swelling, more survival neurons.3. TTC staining was used to observe the morphological changes of brain tissue in general:brain tissue in the sham operation group were stained uniformly red, the remaining rats after modeling, brain size is not a white area, with significant difference(p<0.01), with the extension of time, infarction area increased. Compared with the model group, electroacupun-cture group, blocker group, electroacupuncture+blocker infarcted area reduced in varying degrees,6h after reperfusion in rats was reduced, the area with statistical significance (p<0.05).3d and1w, the infarcted area in the electroacupuncture group, electroacupuncture+blocker than blocker group(p<0.05), the area of infarction+blocker than small electroacupunc-ture EA group (p<0.01).4. TUNEL method to observe the apoptosis of nerve cells:compared with the sham operation group, the number of positive cells increased signifycantly in the other four groups of rats (p<0.01). Positive cells of model group rats showed a time dependent, reached a peak at Id,3d decreased the most, compared with model group, electroacupuncture group, electroacupuncture+blocker, blocker group group of apoptosis index are decreasing, the majority group compared with the model group significantly decreased(p<0.05or p<0.01). Blockers and electroacupuncture combined, apoptosis index decreased significantly, compared with when used alone, with significant difference (p<0.05); individually by using blockers or electroacupuncture stimulation, the difference was statistically significant(p>0.05).5. Comparison of the expression of p-p38MAPK in brain tissue:compared with sham operation group, improve the rest of brain tissue in rats of the four groups acidification p38MAPK protein content(p<0.05). Phosphoryla-tion of p38began in the model group after reperfusion2h increased,1d increased significantly,3d reached the peak, then decreased gradually, the1w positive cell rate is still higher than that in sham operation group. Compared with the model group, the positive rate of electroacupuncture group, electroacupuncture+blocker, blocker group slots for each group were decreased, the difference was significant (p<0.05or p<0.01), and EA group, blocker group and EA+blocker group, the positive cell rate had no signi-ficant difference(p>0.05). Conclusion:1.EA"Chize, Hegu" and "Zusanli, Sanyinjiao" two group of acupuncture point, can improve cerebral ischemia/reperfusion injury in rats neurolo-gical deficits and brain tissue pathological changes, can significantly reduce the apoptosis of nerve cells in ischemic region index, has certain protective action prompted EA"Chize, Hegu" and "Zusanli, Sanyinjiao"two acupoints ischemic injury of brain tissue. 2. Expression of EA"Chize, Hegu" and "Zusanli, Sanyinjiao" can inhibit the phosphorylation of p38MAPK protein, suggesting a protective effect of elec-troacupuncture at"Chize, Hegu" and "Zusanli, Sanyinjiao" on cerebral ische-mia injury may be accomplished by inhibiting the expression of p38MAPK signaling pathway.3. EA"Chize, Hegu"and "Zusanli, Sanyinjiao" on cerebral ischemia/reper-fusion rat neural function defect, brain tissue pathological morphology of nerve cells in ischemic region, apoptosis index and p-p38MAPK improvement in reperfusion after3days when the more obvious, to illustrate the point in time may be to promote better therapeutic time window in brain repair ischemic nerve.4. The experimental results suggested that:"EA stimulation-start p38MAPK signal regulation-antagonist apoptosis"is one of the possible mechanism of acupuncture for treatment of cerebral ischemic injury. |
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