推拿调控局部炎性微环境对腰椎间盘突出症大鼠疼痛行为的影响

Effects of Tuina regulating local inflammatory microenvironment on pain behavior in lumbar disc herniation rats

目的:观察局部炎性微环境与大鼠疼痛行为之间的相关性,探讨推拿干预腰椎间盘突出症(LDH)相关疼痛的可能机制。方法:将32只大鼠随机分为空白组、假手术组、模型组、推拿组,每组8只。推拿组在造模后第4天开始进行推拿干预。实验过程中行大鼠右后足缩足阈值(PWT)测试、热缩足反射潜伏期(PWL)测试及Catwalk步态测试。实验结束后取大鼠背根神经节(DRG)组织,采用HE染色观察大鼠DRG组织病理形态变化;采用ELISA法检测组织中炎症相关因子肿瘤坏死因子(TNF)-α、白细胞介素(IL)-1β、IL-6的表达情况。结果:模型组大鼠造模后至第14天PWT和PWL持续降低,而从第14天至21天的变化趋于稳定,并显著低于同期空白组(P<0.01);推拿组大鼠的PWT和PWL分别从第10天和第7天开始显著高于同期模型组(P<0.01)。与空白组同期比较,模型组大鼠在造模后步态参数显著改变(P<0.01);推拿组大鼠最大接触面积从第7天开始相比同期模型组有显著改善(P<0.05),而站立相持续时间、摆动相持续时间、足印面积、平均强度则是从第14天开始显著改善(P<0.05,P<0.01)。HE染色显示,模型组大鼠的DRG组织内细胞形态发生明显病理改变,并伴有血管充血及炎症细胞浸润;而推拿可使得细胞形态改善,虽然依然有少量炎症细胞,但病理学改变较模型组明显减轻。ELISA结果显示模型组大鼠DRG组织炎症相关因子TNF-α、IL-1β、IL-6的表达显著高于空白组(P<0.01),而推拿后可以显著降低这些因子表达(P<0.01)。结论:局部炎性微环境的改变可能会导致大鼠的疼痛行为,而推拿干预或可通过调控局部炎性微环境来缓解疼痛。

Objective: To observe the relationship between local inflammatory microenvironment and pain behavior in rats, and to explore the possible mechanism of Tuina intervention on pain related to lumbar disc herniation(LDH). Methods:Thirty-two rats were randomly divided into Naive group, Sham group, chronic compression of dorsal root ganglion(CCD) group and Tuina group, 8 rats in each group. Tuina intervention was implemented from the 4th day after surgery in Tuina group. During the experiment, paw withdrawal threshold(PWT) and thermal paw withdrawal latencies(PWL) were measured to observe the changes of mechanical and thermal pain thresholds;Catwalk gait test was conducted to observe the changes of pain related gait parameters. After the experiment, the dorsal root ganglion(DRG) tissues of rats were taken, and the pathological changes of DRG were observed by HE staining;the expressions of tumor necrosis factor(TNF)-α, interleukin(IL)-1β and IL-6 in DRG tissues were detected by ELISA. Results: PWT and PWL of rats in CCD group decreased continuously from the day of surgery to the 14th day, but tended to be stable from the 14th day to the 21st day, and were significantly lower than those in Naive group at the same time point(P<0.01);PWT and PWL of rats in Tuina group were significantly higher than those in CCD group at the same time point from the 10th day and the 7th day respectively(P<0.01). The gait parameters of CCD group were significantly different from Naive group at the same time point(P<0.01);the maximum contact area of rats in Tuina group was significantly improved from the 7th day compared with that of CCD group at the same time point(P<0.05), while the stand duration, swing duration,print area and mean intensity were from the 14th day(P<0.05, P<0.01). HE staining showed that the morphology of DRG cells in CCD group changed significantly, accompanied by vascular congestion and inflammatory cell infiltration;Tuina could improve cell morphology, although there were some inflammatory cells, the pathological changes were significantly less than those in CCD group. The results of ELISA showed that the expressions of TNF-α, IL-1 β and IL-6 in DRG tissues in CCD group were significantly higher than those in Naive group(P<0.01), and Tuina could significantly reduce these factors(P<0.01). Conclusion:The changes of local inflammatory microenvironment may lead to pain behavior in rats, and Tuina intervention may alleviate pain by regulating local inflammatory microenvironment.