主 办：北 京 中 医 药 大 学
ISSN 1006-2157　CN 11-3574/R
北京中医药大学学报

北京中医药大学学报 ›› 2021, Vol. 44 ›› Issue (3): 280-288.doi: 10.3969/j.issn.1006-2157.2021.03.014

• 针灸 • 上一篇    

基于Nrf2/HO-1信号通路探讨艾灸对卵巢储备功能减退大鼠的炎性调控机制*

汪倩, 卢鸽, 李红晓, 时玲, 梁尚杰, 沈梅红#   

  1. 南京中医药大学针灸推拿学院 江苏 210023
  • 收稿日期:2020-07-23 出版日期:2021-03-30 发布日期:2021-03-30
  • 通讯作者: #沈梅红,女,博士,教授,博士生导师,主要研究方向:针灸防治卵巢相关疾病的效应机制,E-mail: mhshen@njucm.edu.cn
  • 作者简介:汪倩,女,在读硕士生
  • 基金资助:
    *国家自然科学基金面上项目(No. 81774408,No. 81973957),江苏省研究生科研创新计划(No.KYCX20_1541)

Inflammatory regulation mechanism of moxibustion on rats with diminished ovarian reserve based on Nrf2/HO-1 signaling pathway*

Wang Qian, Lu Ge, Li Hongxiao, Shi Ling, Liang Shangjie, Shen Meihong#   

  1. School of Acupuncture, Moxibustion and Tuina, Nanjing University of Chinese Medicine, Jiangsu 210023, China
  • Received:2020-07-23 Online:2021-03-30 Published:2021-03-30
  • Contact: Prof. Shen Meihong, Ph.D., Doctoral Supervisor. School of Acupuncture, Moxibustion and Tuina, Nanjing University of Chinese Medicine. No. 138 Xianlin Avenue, Qixia District, Nanjing 210023. E-mail: mhshen@njucm.edu.cn
  • Supported by:
    National Natural Science Foundation of China (No. 81774408, No. 81973957), Research and Innovation Program for Graduate Students in Jiangsu Province (No. KYCX20_1541) Ethical review: Animal Ethics Committee of Nanjing University of Chinese Medicine (No. 201909A027)

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摘要： 目的 探索艾灸对卵巢储备功能减退(DOR)大鼠模型卵巢的保护作用是否与核因子E2相关因子2(Nrf2)/血红素加氧酶1(HO-1)信号通路介导的炎性调节有关。方法 将50只雌性大鼠随机分为空白组、雷公藤多苷(GTW)组、艾灸组、全反式维甲酸(ATRA)组和二甲基亚砜(DMSO)组,每组10只。GTW组、艾灸组、ATRA组和DMSO组均采用雷公藤多苷片混悬液灌胃制备DOR大鼠模型;空白组每日灌胃等体积生理盐水;艾灸组在每日灌胃1h后给予双侧“肾俞”穴或“关元”“中脘”穴艾灸治疗,2组穴隔日交替,每次10min;ATRA组在灌胃30min后腹腔注射ATRA混悬液,30min后同艾灸组进行艾灸干预;DMSO组在灌胃30min后腹腔注射同体积DMSO溶液,其余同ATRA组。每日上午观察大鼠的体重及动情周期;干预结束后,计算大鼠卵巢脏器指数;HE染色观察大鼠卵巢组织学形态结构;ELISA法检测血清中性激素以及炎性因子的变化;免疫组化和PCR分别检测卵巢组织中Nrf2、HO-1蛋白及mRNA的表达情况。结果 与空白组相比,GTW组大鼠卵巢指数明显下降(P<0.05),动情周期紊乱率显著升高(P<0.01),血清卵泡刺激素(FSH)、睾酮(T)、干扰素-γ(INF-γ)水平上升(均P<0.01),抗苗勒管激素(AMH)、白介素4(IL-4)水平下降(均P<0.01),卵巢中Nrf2和HO-1蛋白(均P<0.01)及mRNA(均P<0.05)的表达显著减少;与GTW组相比,艾灸组和DMSO组大鼠卵巢指数呈上升趋势(均P<0.05),动情周期紊乱率明显下降(均P<0.01),血清FSH、T、INF-γ水平减少(均P<0.01),AMH(P<0.01,P<0.05)、IL-4(均P<0.01)水平增加,卵巢中Nrf2和HO-1蛋白及mRNA表达显著增多(均P<0.01);与艾灸组相比,ATRA组大鼠卵巢指数下降(P<0.01),动情周期紊乱率升高(P<0.01),血清FSH、T、INF-γ水平上升(均P<0.01),AMH、IL-4水平下降(P<0.05,P<0.01),卵巢中Nrf2和HO-1蛋白(均P<0.01)及mRNA(均P<0.05)的表达减少。结论 艾灸可通过激活Nrf2/HO-1信号通路减缓GTW所致DOR大鼠的炎症损伤,发挥卵巢保护作用。

关键词: 艾灸, 卵巢储备功能减退, 炎性调控, Nrf2/HO-1信号通路, 大鼠

Abstract: Objective To explore whether the protective effect of moxibustion on ovary of rats with diminished ovarian reserve (DOR) is related to the inflammatory regulation mediated by nuclear factor erythroid2-related factor 2 (Nrf2) /hemeoxygenase1 (HO-1) signaling pathway. Methods Fifty female rats were randomly divided into blank group, multi-glycosides of Tripterygium wilfordii (GTW) group, moxibustion group, all-trans retinoic acid (ATRA) group and dimethyl sulfoxide (DMSO) group, 10 in each group. The GTW group, moxibustion group, ATRA group and DMSO group were treated with Tripterygium wilfordii polyglycoside tablet suspension by gavage to establish DOR rat model. The blank group were given the same amount of normal saline by gavage daily. The moxibustion group was treated by moxibustion at bilateral BL23 on alternate days and CV4 and CV12 on the other days for 10 minutes each time one hour after intragastric administration. The ATRA group received intraperitoneal injection of ATRA suspension 30 minutes after intragastric administration and moxibustion intervention together with the moxibustion group 30 minutes later. The DMSO group received intraperitoneal injection of the same volume of DMSO solution 30 minutes after intragastric administration and the same moxibustion treatment as the ATRA group. The weight and the estrous cycle of rats were observed every morning. After the intervention, the coefficients of ovarian organs of the rats were calculated, the histological structure of ovaries was observed with HE staining, the changes in serum sex hormones and inflammatory factors were detected by ELISA, the expression levels of Nrf2, HO-1 protein and mRNA were detected by immunohistochemistry and PCR respectively. Results Compared with the blank group, the ovarian coefficients of rats in the GTW group decreased significantly (P<0.05), the detection rate of estrous cycle disorder increased significantly (P<0.01), the serum levels of FSH, T, INF-γ increased (all P<0.01), the levels of AMH(P<0.01, P<0.05) and IL-4(all P<0.01) decreased, and the expression levels of Nrf2 and HO-1 protein (all P<0.01) and mRNA (all P<0.05) in ovarian tissues decreased significantly. Compared with the GTW group, the ovarian coefficients of rats in the moxibustion group and the DMSO group increased (all P<0.05), the detection rates of estrous cycle disorder decreased (all P<0.01), the serum levels of FSH, T, INF-γ decreased (all P<0.01), the levels of AMH (P<0.01,p<0.05)and IL-4 increased (all P<0.01), the expression levels of Nrf2 and HO-1 protein and mRNA in ovarian tissues increased significantly (all P<0.01). Compared with the moxibustion group, the ovarian coefficients of rats in the ATRA group decreased (P<0.01), the detection rate of estrus cycle disorder increased (P<0.01), the serum levels of FSH, T, INF-γ increased (all P<0.01), the levels of AMH and IL-4 decreased (P<0.05, P<0.01), the expression levels of Nrf2 and HO-1 protein (all P<0.01) and mRNA (all P<0.05) in ovarian tissues decreased. Conclusion Moxibustion can attenuate the inflammatory injury induced by GTW in DOR rats and protect the ovary by activating Nrf2 / HO-1 signaling pathway.

Key words: moxibustion, diminished ovarian reserve, inflammatory regulation, Nrf2/HO-1 signaling pathway, rats

中图分类号: 

  • R246.3