基于cAMP-PKA信号通路调控TRPV1表达研究温和灸治疗原发性痛经大鼠作用机制  

Study on the Mechanism of Mild Moxibustion in Treating Primary Dysmenorrhea Rats Based on the Regulation of TRPV1 Expression Based on cAMP-PKA Signaling Pathway

作　　者：孙艳秋 梁玉磊 王迪 杨丽芸 张选平 张岩 张璇 邢彦雪 周敏 王晓妍 李新华 SUN Yanqiu;LIANG Yulei;WANG Di;YANG Liyun;ZHANG Xuanping;ZHANG Yan;ZHANG Xuan;XING Yanxue;ZHOU Min;WANG Xiaoyan;LI Xinhua(School of Acupuncture-moxibustion and Tuina,Hebei University of Chinese Medicine,Shijiazhuang 050200,China;Hebei International Joint Research Center for Dominant Diseases in Chinese Medicine and Acupuncture,Shijiazhuang 050200,China)

机构地区：[1]河北中医药大学针灸推拿学院,河北石家庄050200 [2]河北中医针灸优势病症国际联合研究中心,河北石家庄050200

出　　处：《中国中医药信息杂志》2024年第10期96-101,共6页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：河北省自然科学基金(H2022423368、H2023423004)。

摘　　要：目的基于cAMP-PKA信号通路调控瞬时受体电位香草酸亚型1(TRPV1)表达研究温和灸“神阙”“关元”治疗原发性痛经(PD)大鼠的作用机制。方法将32只雌性未孕Wistar大鼠随机分为空白组、模型组、温和灸组和辣椒平组,每组8只。除空白组外,其余组采用腹腔注射苯甲酸雌二醇+冰水浴建立PD寒湿凝滞证大鼠模型。造模第1日开始干预,温和灸组选取“神阙”“关元”温和灸,20 min/次,辣椒平组皮下注射辣椒平2 mg/kg,每日1次,连续10 d。观察大鼠扭体行为并进行评分,ELISA检测子宫组织前列腺素F_(2α)(PGF_(2α))、环磷酸腺苷(c AMP)含量,免疫荧光染色检测子宫组织TRPV1表达,Western blot检测子宫组织蛋白激酶A(PKA)、p-PKA、TRPV1蛋白表达。结果与空白组比较,模型组大鼠扭体潜伏期减少,扭体评分升高(P<0.01);子宫组织PGF_(2α)、cAMP含量升高(P<0.01),TRPV1、p-PKA蛋白表达升高(P<0.01)。与模型组比较,温和灸组和辣椒平组大鼠扭体潜伏期增加,扭体评分降低(P<0.01);子宫组织PGF_(2α)、cAMP含量降低(P<0.01),TRPV1、p-PKA蛋白表达降低(P<0.05,P<0.01)。与温和灸组比较,辣椒平组大鼠扭体潜伏期增加,扭体评分降低(P<0.01);子宫组织PGF_(2α)、cAMP含量降低(P<0.05,P<0.01),TRPV1蛋白表达降低(P<0.05)。结论温和灸“神阙”“关元”对PD大鼠具有明显镇痛作用,其机制可能与调控子宫组织cAMP-PKA信号通路介导TRPV1蛋白表达有关。Objective To explore the mechanism of the treatment of primary dysmenorrhea(PD)by mild moxibustion on“Shenque”and“Guanyuan”acupoints based on the regulation of cAMP-PKA signaling pathway on TRPV1.Methods Totally 32 female non-pregnant Wistar rats were randomly divided into blank group,model group,mild moxibustion group and capsazepine group,with 8 rats in each group.Except for the blank group,the other groups all used estradiol benzoate intraperitoneal injection combined with ice water bath to establish a PD cold-dampness stagnation syndrome rat model.Intervention began on the first day of modeling,the mild moxibustion group selects“Shenque”and“Guanyuan”for mild moxibustion,20 min per time,the capsazepine group was injected capsazepine 2 mg/kg,once a day for 10 consecutive days.ELISA was used to detect uterine PGF_(2α)and cAMP content,immunofluorescence staining was used to detect TRPV1 expression in uterine tissue,Western blot was used to detect PKA,p-PKA and TRPV1 protein expression.Results Compared with the blank group,the latency period of body twisting in the model group rats decreased,and the body twisting score increased(P<0.01);the contents of PGF_(2α)and cAMP in uterine tissue increased(P<0.01),and the expressions of TRPV1 and p-PKA proteins increased(P<0.01).Compared with the model group,the mild moxibustion group and capsazepine group showed an increase in the latency period of body twisting and a decrease in the body twisting score(P<0.01);the content of PGF_(2α)and cAMP in uterine tissue decreased(P<0.01),and the expressions of TRPV1 and p-PKA proteins decreased(P<0.05,P<0.01).Compared with the mild moxibustion group,the capsazepine group showed an increase in the latency period of body twisting and a decrease in the body twisting score(P<0.01);the contents of PGF_(2α)and cAMP in uterine tissue decreased(P<0.05,P<0.01),and the expressions of TRPV1 protein decreased(P<0.05).Conclusion Mild moxibustion at“Shenque”and“Guanyuan”acupoints has obvious analgesic effect on PD rats

关 键 词：原发性痛经 温和灸 cAMP-PKA信号通路 瞬时受体电位香草酸亚型1 大鼠 

分 类 号：R245.8[医药卫生—针灸推拿学]