温针灸对大鼠脊髓损伤节段神经再生抑制因子RhoA、RockⅡ表达的影响

Effects of warm acupuncture and moxibustion on the expression of RhoA and RockⅡin rats with spinal cord injury

目的:观察温针灸对大鼠脊髓损伤(SCI)节段神经再生抑制因子Ras同源物基因A(RhoA)、Rho蛋白激酶Ⅱ(RockⅡ)表达的影响,探讨温针灸治疗脊髓损伤的作用机制。方法:将40只SD大鼠随机分为假手术组、模型组、温针灸组和法舒地尔组,每组10只。假手术组仅去掉椎板,其余三组均以半切法建立SCI模型。假手术组、模型组术后不进行干预,温针灸组造模成功后予温针灸干预,法舒地尔组腹腔注射盐酸法舒地尔抑制剂。HE染色和尼氏染色观察组织形态学变化,免疫组化观察RhoA、RockⅡ蛋白表达,RT-qPCR检测RhoA、RockⅡmRNA表达。结果:假手术组脊髓神经细胞分布均匀、灰质白质分界清楚,尼氏细胞结构完整、数量分布集中、尼氏小体形态正常、着色均匀。模型组组织结构极度紊乱,细胞水肿、炎性浸润明显,尼氏细胞结构模糊、尼氏小体为异常的小颗粒状,着色不均。温针灸组和法舒地尔组均可逆转上述现象,改善组织水肿、增加神经元数量,促进尼氏细胞和尼氏小体组织形态的恢复。干预7、14 d,模型组RhoA、RockⅡ表达均高于假手术组同时间节点(P<0.05);干预7、14 d,温针灸组、法舒地尔组RhoA、RockⅡ表达低于模型组同时间节点,差异有统计学意义(P<0.05)。结论:温针灸可促进SCI组织形态恢复、提高损伤神经再生能力,其作用机制可能与下调脊髓受损节段神经再生抑制因子RhoA、RockⅡ的表达有关。

Objective:To observe the effects of warm acupuncture intervention on the expression of RhoA and RockⅡ,investigate the mechanism of action of warm acupuncture in the treatment of spinal cord injury in rats.Methods:Forty SD rats were randomly divided into sham operation group,model group,warm acupuncture group and fasudil group,with 10 rats in each group.Only lamina was removed in sham operation group,and SCI model was established by half-cut method in the other three groups.The sham operation group and the model group did not receive postoperative intervention,the warm acupuncture group was given warm acupuncture intervention after successful modeling,and the fasudil hydrochloride inhibitor was intraperitoneally injected in the fasudil group.The protein expression of RhoA and RockⅡwas observed by immunohistochemistry.The mRNA expression of RhoA and RockⅡwas detected by RT-qPCR.Results:In the sham operation group,the distribution of spinal cord nerve cells was uniform,the boundary between gray matter and white matter was clear,the structure of Nissl body cells was complete,the number distribution was concentrated,the morphology of Nissl body was normal,and the staining was uniform.In the model group,the tissue structure was extremely disordered,the cell edema and inflammatory infiltration were obvious,the Nissl cell structure was fuzzy,and the Nissl bodies were abnormal small granules with uneven coloring.Warm acupuncture group and fasudil group can reverse the above phenomena,improve tissue edema,increase the number of neurons,and promote the recovery of Nissl cells and Nissl bodies.On day 7 and 14 of intervention,the expressions of RhoA and RockⅡin model group were higher than those in sham operation group at the same time node(P<0.05).On day 7 and 14 of intervention,the expressions of RhoA and RockⅡin warm acupuncture group and fasudil group were lower than those in model group at the same time point,and the differences were statistically significant(P<0.05).Conclusion:Warm acupuncture and moxibustion can promote the recovery of SCI tissue morphology and improve the regeneration ability of injured nerve,and its mechanism may be related to the down-regulation of the expression of RhoA and RockⅡ,which inhibit nerve regeneration in injured spinal cord segments.