摘要
目的 探讨壮医经筋推拿对神经病理性疼痛大鼠去泛素化酶BRCC3介导NOD样受体家族热蛋白结构域相关蛋白3(NLRP3)去泛素化修饰的影响及其镇痛分子学机制。方法 将45只雌性SD大鼠随机分为5组,每组9只。模型组、假手术组、壮医经筋推拿组采用L5脊神经结扎的方法建立神经病理性疼痛模型。于术后第1天开始,假推拿组对大鼠双侧后肢进行轻轻抚摸,壮医经筋推拿组采用按摩推拿手法模拟仪刺激大鼠脊神经结扎节段相应体表部位及所结扎神经支配区域并进行推拿,连续干预14 d;其余3组不予干预,观察14 d。于术前及术后1 d、3 d、14 d测定各组大鼠机械性痛觉阈值和热痛觉阈值;术后14 d,采用qPCR技术检测脊髓背角中BRCC3和NLRP3 mRNA表达情况,采用ELISA法检测血清白细胞介素-1β(IL-1β)水平。结果 术后1 d、3 d,各手术组大鼠的机械性痛觉阈值和热痛觉阈值均明显低于正常组(P均<0.05);术后14 d,壮医经筋推拿组大鼠的机械性痛觉阈值和热痛觉阈值均明显高于模型组和假推拿组(P均<0.05)。术后14 d,模型组和假推拿组大鼠脊髓背角中BRCC3、NLRP3mRNA相对表达量及血清IL-1β水平均明显高于正常组(P均<0.05),壮医经筋推拿组大鼠脊髓背角中BRCC3、NLRP3mRNA相对表达量及血清IL-1β水平均明显低于模型组和假推拿组(P均<0.05)。结论 壮医经筋推拿可减轻神经病理性疼痛大鼠痛敏反应,其机制与下调脊髓背角中去泛素化酶BRCC3表达,以抑制NLRP3的去泛素化修饰,从而阻止NLRP3的活化,阻碍IL-1β成熟和分泌,阻断其介导的免疫炎症反应有关。
Objective It is to investigate the effect of muscle regions massage of Zhuang medicine on the de-ubiquitination modification of Nod-like receptor protein 3(NLRP3)mediated by de-ubiquitinating enzyme BRCC3 in rats with neuropathic pain(NPP)and its molecular mechanism of analgesia.Methods Forty-five female SPF grade SD rats were randomly divided into 5 groups:model group,normal group,group of muscle regions massage of Zhuang medicine,sham massage group and sham operation group,with 9 rats in each group.The models of NPP were established by L5 spinal nerve ligation in the model group,group of muscle regions massage of Zhuang medicine and sham operation group.Beginning on the first postoperative day,the rats of the sham massage group were gently stroked on the bilateral hind limbs,and the rats of the group of muscle regions massage of Zhuang medicine were treated with a massage acupressure manipulation simulator to stimulate the corresponding body surface parts of the spinal nerve ligated segments and the innervation areas of the ligated nerves and with massage,continuously treated for 14 days,while the remaining three groups were given no intervention and were observed for 14 days.The mechanical pain threshold and heat pain threshold of rats in each group were measured before operation and at 1,3,and 14 days after operation;at 14 days after operation,the expressions of BRCC3 and NLRP3 mRNA in the dorsal horn of the L4-6 spinal cord were detected by qPCR,and the serum level of interleukin-1β(IL-1β)was detected by ELISA.Results At 1 and 3 days after operation,the mechanical pain threshold and heat pain threshold of rats in each surgical group were significantly lower than those in the normal group(all P<0.05);at 14 days after operation,the mechanical pain threshold and heat pain threshold of rats in the group of muscle regions massage of Zhuang medicine were significantly higher than those in the model group and sham massage group(all P<0.05).At 14 days after operation,the relative expressions of BRCC3 and NLRP3 mRNA in dorsal horn of spinal cord and levels of IL-1βin serum of rats in the model group and sham massage group were significantly higher than those in the normal group(all P<0.05),and the relative expressions of BRCC3 and NLRP3 mRNA in dorsal horn of spinal cord and levels of IL-1βin serum of rats in the group of muscle regions massage of Zhuang medicine were significantly lower than those in the model group and sham massage group(all P<0.05).Conclusion Muscle regions massage of Zhuang medicine can reduce the pain sensitivity of rats with neuropathic pain,and its mechanism is related to the down-regulation of the expression of the de-ubiquitination enzyme BRCC3 in the dorsal horn of spinal cord,so as to inhibit the de-ubiquitination modification of NLRP3,thus to reduce the activation of NLRP3 and block maturity and secretion of IL-1βto block its mediated immune inflammatory reaction.
作者
王天翊
唐宏亮
梁英业
庞军
卢栋明
唐业妮
郑玉娇
何育风
WANG Tianyi;TANG Hongliang;LIANG Yingye;PANG Jun;LU Dongming;TANG Yeni;ZHENG Yujiao;HE Yufeng(Guangxi University of Traditional Chinese Medicine,Nanning 530000,Guangxi,China;Fangchenggang Hospital of Traditional Chinese Medicine,Fangchenggang 538000,Guangxi,China;The First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine,Nanning 530000,Guangxi,China)
出处
《现代中西医结合杂志》
CAS
2024年第1期17-21,27,共6页
Modern Journal of Integrated Traditional Chinese and Western Medicine
基金
广西自然科学基金项目(2023GXNSFDA026012,2020GXNSFBA297053)
国家自然科学基金资助项目(82205305,82074574,82160943,82260976)。
