壮医药线点灸对带状疱疹后遗神经痛模型大鼠中枢神经系统Th1/Th2/Th17细胞平衡的影响

To Explore the Effect of Zhuang Medicated Thread Moxibustion on Th1/Th2/Th17 Cell Balance in Central Nervous System of Postherpetic Neuralgia Model Rats

目的探究壮医药线点灸对带状疱疹后遗神经痛(Postherpetic neuralgia,PHN)大鼠的干预效果及对中枢神经系统中Th1/Th2/Th17细胞平衡的影响。方法将50只SD大鼠随机分为空白组、模型组、阴性对照组(空线点灸组)、阳性对照组(普瑞巴林组)和壮医药线点灸组,每组10只。除空白组外均予以单次腹腔注射树脂毒素(Resiniferatoxin,RTX)诱导PHN大鼠模型,造模成功后,各组给予相应处理21天,于造模前3天、造模后第1、4、7天以及干预后第3、7、14、21天检测大鼠的缩足机械痛阈值(Mechanical pain threshold,PWT)和大鼠热缩足反射潜伏期(Thermal withdrawal latency,TWL)。干预结束后,取脊髓组织,检测Th1/Th2/Th17细胞比例变化及γ干扰素(Interferon-γ,IFN-γ)、白细胞介素-2(Interleukin-2,IL-2)、白细胞介素-4(Interleukin-4,IL-4)、白细胞介素-10(Interleukin-10,IL-10)、白细胞介素-17(Interleukin-17,IL-17)、白细胞介素-22(Interleukin-22,IL-22)mRNA及蛋白表达情况,阐明该疗法对此平衡机制的调控作用。结果树脂毒素造模后大鼠的PWT呈下降趋势,而TWL呈上升趋势,表现出热痛分离,契合PHN的临床表现。壮医药线点灸后,PHN大鼠PWT上升、TWL下降,改善了热痛分离趋势;阴性对照组、阳性对照组和壮医药线点灸组均能通过促进中枢神经系统中Th1的应答,升高脊髓组织中CD4+IL-2+细胞比例以及IFN-γ、IL-2的含量,上调IFN-γ、IL-2 mRNA的表达以及其蛋白活性,同时,抑制中枢神经系统中Th2、Th17的应答,降低脊髓组织中CD4+IL-4+、CD4+IL-17A+细胞比例以及IL-4、IL-10、IL-17、IL-22的含量,下调IL-4、IL-10、IL-17、IL-22 mRNA的表达以及其蛋白活性,且阳性对照组与壮医药线点灸组在纠正中枢神经系统Th1/Th2/Th17淋巴细胞失衡方面疗效相当,均优于阴性对照组。结论壮医药线点灸可改善带状疱疹后遗神经痛大鼠热痛分离趋势,这可能是通过调节Th1/Th2/Th17细胞平衡及相关细胞因子水平来实现的。

Objective To investigate the effect of Zhuang medicated thread moxibustion on postherpetic neuralgia model rats,and to verify the mechanism of Th1/Th2/Th17 cytokine balance regulation in central nervous system.Methods 50 SD rats were randomly divided into blank group,model group,thread-moxibustion without drugs group,Zhuang medicated thread moxibustion group and pregabalin group,with 10 rats in each group.Except for blank group,the rat model of postherpetic neuralgia was induced by a single intrapitoneal injection of resin-toxin(RTX).After successful modeling,each group was given corresponding treatment for 21 days.The Mechanical Pain Threshold(PWT)and Thermal withdrawal latency(TWL)were detected 3 days before modeling,1,4 and 7 days after modeling,and 3,7,14 and 21 days after intervention.After the intervention,extract spinal cord tissue,Flow cytometry was used to detect changes in the proportion of Th1/Th2/Th17 cells;The mRNA expression levels of IFN-γ,IL-2,IL-4,IL-10,IL-17 and IL-22 were detected by qPCR;The positive expressions of IFN-γ,IL-2,IL-4,IL-10,IL-17 and IL-22 were observed by immunohistochemistry.The protein expressions of IFN-γ,IL-2,IL-4,IL-10,IL-17 and IL-22 were detected by Western blot to elucidate the regulation effect of this therapy on the equilibrium mechanism.Results After intrapitoneal injection of resin-toxin,the PWT of rats showed a downward trend,while the TWL showed an upward trend,showing the separation of heat and pain,which was consistent with the clinical manifestations of PHN.After Zhuang medicated thread moxibustion therapy,PWT increased and TWL decreased in PHN rats,which reduced the separation trend of heat and pain.Thread-moxibustion without drugs group,Zhuang medicated thread moxibustion group and pregabalin group can promote Th1 response in the central nervous system(spinal cord),increase the proportion of CD4+IL-2+cells and the contents of IFN-γand IL-2 in the spinal cord tissue,and up-regulate the expression of IFN-γand IL-2 mRNA and protein.Meanwhile,Th2 and Th17 responses in the central nervous system were inhibited,and the proportions of CD4+IL-4+and CD4+IL-17A+cells and the contents of IL-4,IL-10,IL-17 and IL-22 in the spinal cord tissue were reduced.The mRNA and protein expressions of IL-4,IL-10,IL-17 and IL-22 were down-regulated.In addition,Zhuang medicated thread moxibustion group and pregabalin group had similar efficacy in correcting Th1/Th2/Th17 lymphocyte imbalance in the central nervous system,and both were better than the Thread-moxibustion without drugs group.Conclusion Zhuang medicated thread moxibustion can improve the separation trend of heat and pain in postherpetic neuralgia rats,which may be achieved by regulating Th1/Th2/Th17 cell balance and related cytokine levels.