摘要
目的:基于Toll样受体(TLR)4/髓样分化因子(MyD)88/核转录因子κB p65(NF-κB p65)信号通路探讨石墨烯暖宫穴位贴对原发性痛经(PD)模型大鼠免疫功能的影响及对辅助性T淋巴细胞(Th)1/Th2免疫平衡的调节作用。方法:将SD雌性大鼠随机分成正常组、模型组和穴贴组,每组10只。股部皮下注射苯甲酸雌二醇制备PD模型。在造模的同时予穴贴组大鼠石墨烯暖宫穴位贴贴敷“关元”“子宫”(双侧)和“三阴交”(双侧)干预,连续10 d、1次/d、5 h/次。第11天腹腔注射缩宫素观察各组大鼠的扭体潜伏时间、30 min内的扭体次数,进行扭体评分;计算脾脏和胸腺的脏器指数;HE染色法观察大鼠脾脏和胸腺组织的病理变化;ELISA法检测血清免疫球蛋白(Ig)A、IgG、肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-2、干扰素-γ(IFN-γ)、IL-4和IL-10含量;Western blot法和实时荧光定量PCR法分别检测大鼠子宫组织中TLR4、MyD88、NF-κB p65的蛋白及mRNA的相对表达量。结果:与正常组比较,模型组大鼠出现扭体反应(P<0.01),30 min内扭体次数和扭体评分显著增加(P<0.001);脾脏指数和胸腺指数均显著降低(P<0.01,P<0.05);脾脏和胸腺均出现了明显的病理变化;血清IgA、IgG、TNF-α、IL-2、IFN-γ含量显著升高,IL-4和IL-10含量显著降低(P<0.001,P<0.01);子宫组织TLR4、MyD88、NF-κB p65蛋白及mRNA表达均显著升高(P<0.001)。与模型组比较,穴贴组大鼠扭体潜伏时间延长(P<0.001),其他指标均逆转(P<0.001,P<0.01,P<0.05)。结论:石墨烯暖宫穴位贴可通过调控TLR4/MyD88/NF-κB p65信号通路,调节Th1/Th2的免疫平衡,修复免疫组织的病理损伤,改善免疫功能,从而有效缓解PD大鼠的疼痛症状。
Objective To investigate the effects of graphene-based warm uterus acupoint paste on uterine Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear transcription factor-kappa B p65(NF-κB p65) signaling pathway and Th1/Th2 immune balance in primary dysmenorrhea( PD) model rats, so as to reveal its immunological mechanisms of relieving dysmenorrhea. Methods Thirty SD female rats were randomly divided into 3groups: normal group, model group and acupoint paste group, with 10 rats in each group. PD rat model was established by subcutaneous injection of estradiol benzoate for 10 consecutive days. At the same time of modeling, graphene-based warm uterus acupoint paste was applied to the acupoints of “Guanyuan”(CV4), bilateral “Zigong”(EX-CA1) and “Sanyinjiao”(SP6) of rats in the acupoint paste group. The application was continuously applied once daily for 10 d, 5 h each time. On the 11^(th) day, oxytocin was injected intraperitoneally to observe the writhing latency, writhing times within 30 min and writhing score of rats in each group. The spleen and thymus indexes were calculated.The pathological changes of spleen and thymus tissue were observed after HE staining. The contents of serum immunoglobulin(Ig) A, IgG, tumor necrosis factor-α(TNF-α), interleukin(IL)-2, interferon-γ(IFN-γ), IL-4 and IL-10 were detected by ELISA. The protein and mRNA expression levels of TLR4, MyD88 and NF-κB p65 in rat uterine tissue were detected by Western blot and real-time quantitative PCR, respectively. Results Compared with the normal group, the writhing times and writhing scores within 30 min of rats in the model group were significantly increased(P<0.001), and the rats showed writhing reaction(P<0.01). The spleen index and thymus index were significantly decreased(P<0.01,P<0.05). The spleen and thymus had obvious pathological changes. The contents of IgA, IgG, TNF-α, IL-2 and IFN-γin serum were significantly increased, while the contents of serum IL-4 and IL-10 were significantly decreased(P<0.001,P<0.01). The expression levels of TLR4, MyD88, NF-κB p65 protein and corresponding mRNA in uterine tissue were significantly increased(P<0.001). Following intervention, compared with the model group, the writhing latency time of rats in the acupoint paste group was prolonged, and the writhing times and writhing scores within 30 min were significantly decreased(P<0.001). The spleen index and thymus index were significantly increased(P<0.01,P<0.05).The pathological changes of spleen and thymus were improved. The contents of serum IgA, IgG, TNF-α, IL-2 and IFN-γwere significantly decreased, while the contents of IL-4 and IL-10 were significantly increased(P<0.001,P<0.05,P<0.01). The expression of TLR4, MyD88, NF-κB p65 protein and the corresponding mRNA levels in uterine tissue were decreased(P<0.001, P<0.01). Conclusion Graphene-based warm uterus acupoint paste can regulate the immune balance of Th1/Th2 by regulating TLR4/MyD88/NF-κB p65 signaling pathway, repair the pathological damage of immune tissue, improve immune function, and effectively relieve the pain symptoms of PD rats.
作者
王琳
李铁
曹文轩
赵晋莹
徐晓红
柴佳鹏
张嘉勋
刘佳
王富春
WANG Lin;LI Tie;CAO Wen-xuan;ZHAO Jin-ying;XU Xiao-hong;CHAI Jia-peng;ZHANG Jia-xun;LIU Jia;WANG Fu-chun(College of Acupuncture-moribustion and Tuina,Changchun University of Chinese Medicine,Changchun 130117,China;College of Pharmacy,Changchun University of Chinese Medicine,CChangchun 130117)
出处
《针刺研究》
CAS
CSCD
北大核心
2024年第1期37-46,共10页
Acupuncture Research
基金
国家重点研发计划项目(No.2021YFE0202900)
吉林省科技发展计划项目(No.20210203076SF)。
