摘要
目的:观察针刺背俞穴对失眠大鼠下丘脑中白细胞介素-17(IL-17)、白细胞介素-22(IL-22)、肿瘤坏死因子-α(TNF-α)、辅助性T细胞17(Th17)与调节性T细胞(Treg)、信号转导和转录激活因子3(STAT3)的影响,探讨其改善失眠的机制。方法:将48只SD大鼠随机分为空白组、模型组、西药组和针刺组,每组12只。除空白组外,另外3组均采取腹腔注射对氯苯丙氨酸(PCPA)混悬液制造失眠模型。成模后西药组大鼠灌胃艾司唑仑溶液治疗1周;针刺组采取针刺大鼠双侧五脏背俞穴治疗1周,其他两组仅抓取和固定,不治疗。成模后治疗前,比较各组大鼠的一般情况及睡眠时间变化;治疗结束后,ELISA检测下丘脑及血清中IL-17、IL-22及TNF-α含量;流式细胞术检测下丘脑Th17及Treg阳性细胞率;Western blot检测下丘脑p-STAT3、STAT3蛋白表达。结果:造模后的3组大鼠与空白组比较,睡眠潜伏期变长,差异具有统计学意义(P<0.01),睡眠时间变短,差异具有统计学意义(P<0.01);治疗后与空白组比较,模型组大鼠下丘脑及血清中的IL-17、IL-22及TNF-α含量均升高,差异具有统计学意义(P<0.01),下丘脑中Th17阳性细胞率增加,差异具有统计学意义(P<0.05),Treg阳性细胞率减少,差异具有统计学意义(P<0.05),下丘脑中p-STAT3蛋白表达升高,差异具有统计学意义(P<0.01);与模型组比较,两治疗组大鼠下丘脑及血清中IL-17、IL-22及TNF-α含量均下降,差异具有统计学意义(P<0.01),下丘脑Th17阳性细胞率减少,差异具有统计学意义(P<0.05),Treg阳性细胞率增加(P<0.05,P<0.01),下丘脑中p-STAT3表达下降,差异具有统计学意义(P<0.01);与西药组比较,针刺组大鼠下丘脑及血清中IL-17、IL-22及TNF-α含量下降更多,差异具有统计学意义(P<0.01),Treg阳性细胞率增加更多,差异具有统计学意义(P<0.05)。结论:针刺背俞穴可以下调下丘脑IL-17、IL-22及TNF-α,平衡Th17和Treg细胞含量,调节STAT3信号通路的过度磷酸化,降低其活性,改善下丘脑神经炎症,从而治疗失眠。
Objective:To observe the effects of needling back-Shu points on interleukin-17(IL-17),interleukin-22(IL-22),tumor necrosis factor-α(TNF-α),helper T cell 17(Th17),regulatory T cell(Treg)and signal transduction and transcriptional activator 3(STAT3)in hypothalamus of insomnia rats,thus to explore its mechanism in improving insomnia.Methods:48 SD rats were randomly divided into the blank group,the model group,the Western medication group and the acupuncture group,with 12 rats in each group.The insomnia model was established by intraperitoneal injection of p-chlorophenylalanine(PCPA)suspension.After successful modeling,the Western medication group was treated with Estazolam Solution by gavage for a week,the acupuncture group was treated by needling back-Shu points of bilateral five-Zang for a week,and the other two groups were only intervened by grasping and fixing.Post-modeling and pre-treatment,the general condition and sleep duration were compared among each group.After the treatment,the levels of IL-17,IL-22 and TNF-αin hypothalamus and serum were detected by ELISA.Flow cytometry was used to detect Th17 and Treg positive cells in hypothalamus.The protein expressions of p-STAT3 and STAT3 in hypothalamus were detected by Western blot.Results:Compared with that in the blank group,the sleep latency was lengthened(P<0.01),and the sleep duration was shortened in the other three groups(P<0.01).Compared with those in the blank group,the contents of IL-17,IL-22 and TNF-αin hypothalamus and serum were increased(P<0.01),the rate of Th17 positive cells in hypothalamus was increased(P<0.05),the rate of Treg positive cells was decreased(P<0.05),and the protein expression of P-STAT3 was increased in the model group(P<0.01).Compared with those in the model group,the contents of IL-17,IL-22 and TNF-αin hypothalamus and serum were decreased(P<0.01),the rate of Th17 positive cells in hypothalamus was decreased(P<0.05),the rate of Treg positive cells was increased(P<0.05,P<0.01),and the protein expression of P-STAT3 was decreased in the Western medication group and the acupuncture group(P<0.01).Compared with those in the Western medication group,the contents of IL-17,IL-22 and TNF-αin hypothalamus and serum were significantly decreased(P<0.01),and the rate of Treg positive cells was greatly increased in the acupuncture group(P<0.05).Conclusion:Needling back-Shu points can down-regulate the levels of IL-17,IL-22 and TNF-αin hypothalamus,balance the content of Th17 and Treg cells,regulate the hyperphosphorylation of STAT3 signaling pathway,reduce its activity,and improve the hypothalamic neuroinflammation to treat insomnia.
作者
钱佳慧
高希言
祁尚文
李玉冰
李余洁
李薇
QIAN Jiahui;GAO Xiyan;QI Shangwen;LI Yubing;LI Yujie;LI Wei(Henan University of Chinese Medicine,Zhengzhou 450008,China)
出处
《针灸临床杂志》
2024年第12期72-78,共7页
Journal of Clinical Acupuncture and Moxibustion
基金
河南省中医药科学研究专项课题,编号:2022ZY1096
河南省中医学“双一流”创建科学研究专项课题,编号:HSRP-DFCTCM-2023-3-14。
作者简介
钱佳慧(1995-),女,2021级针灸推拿学专业硕士研究生,研究方向:针灸理论的临床和基础研究;通讯作者:高希言(1962-),男,教授,博士研究生导师,研究方向:针灸理论的临床和基础研究。
