| Purpose:This experiment aims at the study of acupuncture at Huantiao point and Jiaji points different curative effects of sciatic nerve crush injury in rats. With immunohistochemistry and Western blot detection method were observed in rat dorsal root ganglion of PKA and CREB and c-fos content, explore the mechanism of acupuncture in treatment of rat sciatic nerve injury of c AMP signal transduction pathway. Comparing with the effects of Huantiao point and Jiaji points for repair of sciatic nerve injury.Material and method:48 SD healthy rats, male and female, were divided into four groups according to the random number table method: blank group, model group, Huantiao group,Jiaji group, the group of 4 groups, each group of 12. Designed by Hasegawa[1], Liao Weijing [2] and Wang [3] modified clamping method to make the sciatic nerve injury model. Blank group and model group under the same conditions, normal feeding, eighth days after modeling, the rats were fixed on the fixed device, 15 min each time, 1 times a day for 14 days. Eighth days after modeling, acupuncture Huantiaopoints and Jiaji points. Connection SDZ- II electro acupuncture instrument for treatment, each treatment 15 min, once a day, continuous treatment for 14 days. At the end of twenty-second days after modeling, behavioral observation was made to evaluate the muscle strength and motor function recovery in rats by testing the function index of sciatic nerve. In rats with left hip junction of about 1.5 cm long incision incision, exposed the sciatic nerve, the interception of the injury on the 5 mm. And then, the rat dorsal midline longitudinal skin incision, open canal, the interception of L4-L5 segment spinal cord. Morphological observation by HE staining observation of sciatic nerve pathological changes; using molecular biological assay, rat L4-L5 spinal cord and dorsal root ganglion region of PKA, CREB and c-fos expression intensity was observed. The results were statistically analyzed.Results:1.SFIThe functional recovery of the affected limb was observed with the rat sciatic nerve function index. 8 days after modeling, compared with the blank group, sciatic nerve crush model rats of SFI was significantly lower than that of the blank group, the difference was statistically significant(p < 0.05), indicating that the motor function of rats decreased and the modeling. After twenty-second days, compared with model group, there was a significant increase in both the Huantiao group and the Jiaji group, the difference was statistically significant(p<0.05), which showed that acupuncture treatment can promote the recovery and regeneration of sciatic nerve injury in rats. The SFI of sciatic nerve of the Huantiao groupwas superior to that of Jiaji group, and the difference was statistically significant(p<0.05).2.HE stainingRat sciatic nerve injury of HE staining showed: blank group sciatic nerve axons, myelin a good continuity and fiber arranged regularly; compared with the blank group, model group of nerve fibers distributed irregularly, axons, myelin ulceration, partially collapsed; Huantiao group and Jiajigroup sciatic nerve fiber distribution comparison rules, part of the nerve appeared new growth, partial loss of myelin, the Huantiao group was better than that of Jiaji group.3.Immunohistochemical detectionPKA: blank group PKA in L4-L5 dorsal root ganglion are abundantly expressed in and positive cell nucleus is blue, cytoplasm was brown; blankgroup and compared to root ganglion PKA protein expression levels were significantly reduced(p<0.01) dorsal remaining rats L4-L5; compared with the model group, the blank group, Huantiao group, Jia Ji group significantly increased(p<0.01); and Huantiao group comparison, Jiaji group decreased, the difference is statistically significant(p<0.05).CREB: blank group in CREB in L4-L5 dorsal root ganglion are abundantly expressed in, positive cell nuclei showed blue, cytoplasm was brown; blank group and compared to root ganglion of CREB protein expression levels were significantly reduced(p<0.01) dorsal remaining rats L4-L5; compared with the model group, the blank group, Huantiao group, Jia Ji group significantly increased(p<0.01); and Huantiao group comparison, Jiaji group decreased, the difference is statistically significant(p< 0.05).c-fos: c-fos in the blank group at the L4-L5 dorsal root ganglion is expressed in a very small amount, compared with the blank group, root ganglion in c-fos protein expression level significantly increased(p< 0.01) dorsal remaining rats L4-L5; compared with the model group, the blank group, Huantiao group, Jiaji group down regulated significantly(p<0.01); and Huantiao group comparison, Jiajigroup increased, the difference is statistically significant(p<0.05).4.Western Blot detectionPKA: PKA in the blank group at the L4-L5 dorsal root ganglion are abundantly expressed in. Compared with the blank group, model group, Jiajigroup rat L4-L5 dorsal root ganglion PKA protein expression levels were significantly reduced(p<0.01);compared with the model group,the blank group, Huantiaogroup, Jiaji group increase significantly(p<0.01);and Huantiao group comparison, Jiaji group decreased significantly, the difference is statistically significant(p<0.01).CREB: blank group in CREB in L4-L5 dorsal root ganglion are abundantly expressed in, compared with the blank group, root ganglion of CREB protein expression levels were significantly reduced(p<0.05) dorsal remaining rats L4-L5; compared with the model group, the blank group, Huantiao group, Jiaji group significantly increased(p<0.05);and Huantiao group comparison,Jiaji group decreased,the difference is statistically significant(p<0.05).c-fos: c-fos in the blank group at the L4-L5 dorsal root ganglion is expressed in a very small amount, compared with the blank group, model group, Jiajigroup rat L4-L5 dorsal root ganglion of c-fos protein expression level significantly increased(p<0.01); compared with the model group, the blank group, Huantiao group, Jiaji group decreased significantly(p< 0.01); and Huantiao group comparison, Jiaji group increased, the difference is statistically significant(p<0.05).Conclusion:1.Acupuncture Huantiao points and Jiaji points can promote the repair and regeneration of injured sciatic nerve in different degrees.2.Acupuncture Huantiao point and Jiaji points can significantly improve the back root ganglion PKA and CREB content and activating the c AMP pathway of apoptosis, and inhibit the expression of c-fos in, which may be promoting acupuncture is one of effective mechanisms for the repair and regeneration of sciatic nerve injury. |
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