| Objective:By research the effect of acupuncture and neural stem cell combination therapy on hypoxic-ischemic brain damage (HIBD) rats'overall function, apoptosis and anti-apoptotic protein expression and brain tissue pathological ,to study the therapy mechanism of acupuncture and neural stem cell on cerebral palsy and provide theoretical evidence for clinical treatment on the patients with CP.Research contents and methods:1. Experimental animals and group: 7-day SD newborn rats(weight:12g～20g) divided into group①:normal control group randomly. The remaining rats operated to build HIBD model, the successful model was divided into group②:model control group, group,③:acupuncture group, group④:NSC group, group⑤:acupuncture + NSC group randomly by block. 12 rats included in each group.2. Treatment stage: Including acupuncture therapy and neural stem cell therapy, neural stem cell by intravenous injection.3.Behavior detection:including over hanging test,inclined plane test,Y maze test.4. Draw the materials: drawn the normal group and experimental group of rats by cervical 21 days after the treatment. Observe pathological changes in hippocampus by using HE staining;Detect nerve cell cell apoptosis by using TUNEL. Detect bcl-2 and bax protein expression in brain by the protein chip. 5. Data management, statistical analysis.Results:1. Behavior detection:in the tests of over hanging test,inclined plane test,Y maze test, compare②m odel group with①normal group, the difference was statistical significant (p<0.05), Show that the model of HIBD was made successfully; Compare②model group with③a cupuncture group, the result was no significant difference (p >0.05); Compare②model group with④NSC group, the result was no significant difference also(p>0.05); Show that pure NSC or acupuncture treatment can not improve motor function of limbs and intelligence in young rats significantly; Compare②model group with⑤acupuncture + NSC group, the difference was statistical significant (p<0.05); Show that⑤acupuncture + NSC group can improve motor function of limbs and intelligence in young rats significantly.2. Protein express of bcl-2 and bax gene: Compared⑤acupuncture and NSC group with②model group, the mean difference was statistically significant (p<0.05), showed that combination treatment with acupuncture and NSC could promote anti-apoptotic gene bcl-2 protein expression and inhibit the expression of apoptotic genes bax protein; Compared③simple acupuncture group or④NSC group with②model group,there are no significant difference (p>0.05).3. Brain tissue in general: Group③appeared large brain area tissue necrosis and vacuoles, The structure of right brain tissue is normal. The left side of brain tissue have suffered different degrees of necrosis and cavitation region in the group of③acupuncture group,④NSC group,⑤acupuncture + NSC group,and the level of damage is slighter; but the level of damage in group⑤acupuncture + NSC is slightest.4. Brain tisue test by HE staining: Nerve cells of degeneration in hippocampus reduced significantly in group③acupuncture group,④NSC group and⑤acupuncture + NSC group compared with the②model group. there is a significant improvement in the Pathomorphology. The number of nerve cells with degeneration is minimum and brain lesions lightest in the group⑤acupuncture + NSC.5. Nerve cell counts: (1) the number of normal hippocampal neurons : compare②model group with①normal group, the difference was statistical significant (p <0.05), Show that the number of normal nerve cells in Hippocampus in rats reduced significantly after HIBD. Compared②model group with③a cupuncture group or④NSC group the result have no significant difference (p>0.05),show that③acupuncture group or④NSC group cannot improve the counts of neve cell in hippocamous. compared②model group with⑤acupuncture + NSC group the result have significant difference (p<0.05),show that⑤acupuncture+NSC group can improve the counts of neve cell in hippocamous.(2)Apoptosis of cortical cells: compared②model group with③a cupuncture group and④NSC group the result have no significant difference(p>0.05); compared②model group with⑤acupuncture + NSC group the result have significant difference (p<0.05),show that⑤acupuncture + NSC group can improve the counts of neve cell in hippocamous.and can inhibit the apoptosis of cells in cortical of rats obviously.Conclusion:1. Treatment with only acupuncture or neural stem cell cannot obviously effect on weight gain,muscle growth,coordination of recovery,learning and memory (mental) recovery and cannot inhibit protein expression of apoptosis gene bax and can not stimulate promote protein expression of anti-apoptosis gene bcl-2 in rats of HIBD CP model.2.There are significant effect on weight gain,muscle growth,coordination of recovery,learning and memory (mental) recovery in rats of HIBD CP model. by combination treatment of acupuncture and neural stem cell.3. Combination treatment of acupuncture and Neural stem cell can inhibit protein expression of apoptosis gene bax and promote protein expression of anti-apoptosis gene bcl-2 in HIBD CP model rats obviously. |
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