摘要
目的探讨针刺对卵巢储备功能减退模型大鼠动情周期的影响及可能作用机制。方法将40只动情周期正常的SD雌性大鼠随机分为空白组、模型组、针刺组、激素组,每组10只。除空白组外,其余3组大鼠予雷公藤多苷片混悬液按50mg/(kg·d)连续灌胃14天建立卵巢储备功能减退大鼠模型。针刺组在每日雷公藤多苷灌胃1h后进行针刺干预,选取"肾俞""关元""中脘"穴,每次行针10s,共留针10min。同时激素组进行雌激素、孕激素序贯疗法。实验第8天起观察大鼠动情周期,第15天采用HE染色法观察各组大鼠卵巢中卵泡、黄体发育情况;检测大鼠血清激素[包括抗苗勒管激素(AMH)、雌二醇(E2)、卵泡刺激素(FSH)]及炎性因子[包括肿瘤坏死因子α(TNF-α)、白细胞介素21(IL-21)、白细胞介素10(IL-10)]水平;检测卵巢组织中转录因子核因子E2相关因子2(Nrf2)、血红素氧合酶1(HO-1)蛋白表达及Nrf2、HO-1、核苷酸结合寡聚化结构域受体3(NLRP3)mRNA相对表达量。结果与空白组比较,模型组大鼠的动情周期延长或紊乱,生长卵泡颗粒细胞层减少,黄体萎缩,闭锁卵泡和炎性浸润增多;血清AMH、E2、IL-10水平降低,FSH、TNF-α、IL-21水平升高,卵巢组织中Nrf2、HO-1蛋白和mRNA表达降低,NLRP3 mRNA升高(P<0.01)。与模型组比较,针刺组及激素组生长卵泡颗粒细胞层增多,闭锁卵泡和炎性浸润减少;血清AMH、E2、IL-10升高,FSH、IL-21下降;卵巢组织中Nrf2、HO-1蛋白和mRNA增高,NLRP3 mRNA降低(P<0.05或P<0.01)。针刺组和激素组各指标差异无统计学意义(P>0.05)。结论针刺能够恢复卵巢储备功能减退模型大鼠的动情周期,有效调节大鼠性激素水平,可能通过Nrf2/HO-1信号通路抑制NLRP3 mRNA表达,从而抑制炎症反应。
Objective To explore the effects and possible mechanism of acupuncture on the estrous cycle of rat model of hypo-ovarian reserve function. Methods A total of 40 SD female rats with normal estrus cycle were randomly divided into a blank group, a model group, an acupuncture group and a hormone group, with 10 rats in each group. Except for the blank group, rats were given tripterygium wilfordii polyglycoside tablet suspension at 50 mg/(kg·d) for 14 days to establish hypo-ovarian reserve dysfunction model rat. In the acupuncture group, acupuncture intervention was performed one hour after gavage of tripterygium wilfordii polyglycoside every day, and the acupoints of "Shenshu"(BL23), "Guanyuan"(CV4) and Zhongwan(CV12) were selected, the needling was given for 10 s each time with retension for 10 minutes. Meanwhile, the hormone group was given sequence therapy of estrogen and progesterone. The estrous cycle of rats was observed from the 8 th day of the experiment. On the 15 th day. HE staining method was used to observe the development of follicles and corpus luteum in the ovaries of each group of rats;serum hormones including anti-Müllerian hormone(AMH), estradiol(E2), follicle stimulating hormone(FSH) and inflammatory factors including tumor necrosis factor alpha(TNF-α), interleukin 21(IL-21), interleukin 10(IL-10) levels were detected. The level of transcription factor nuclear factor E2 related factor 2(Nrf2), heme oxygenase 1(HO-1) protein expression and Nrf2, HO-1, nucleotide binding oligomerization domain receptor 3(NLRP3) relative mRNA expression quantity were detected. Results Compared with the blank group, the model group had prolonged or disordered estrous cycle, decreased granulosa cell layer of growth follicle, atrophy of corpus luteum, increased atresia follicle and inflammatory infiltration;serum AMH, E2, IL-10 levels decreased, the levels ofFSH, TNF-α and IL-21 increased, the expression of Nrf2, HO-1 protein and mRNA in ovarian tissue decreased, and the NLRP3 mRNA increased(P<0.01). Compared with the model group, the growth follicle granular cell layer increased in the acupuncture group and the hormone group, the atresia follicles and inflammatory infiltration decreased;serum AMH, E2, IL-10 increased, FSH, IL-21 decreased;Nrf2, HO-1 and mRNA protein in ovarian tissue increased, NLRP3 mRNA decreased(P<0.05 or P<0.01). There was no significant difference in all indicators between the acupuncture group and the hormone group(P>0.05). Conclusion Acupuncture can restore the estrous cycle of rats with hypo-ovarian reserve function and effectively regulate the level of sex hormones in rats. The action may be related to inhibition of the NLRP3 mRNA expression by Nrf2/HO-1 signaling pathway to inhibit the inflammatory response.
作者
卢鸽
解子婧
汪倩
李茜
李红晓
沈梅红
LU Ge;XIE Zijing;WANG Qian;LI Qian;LI Hongxiao;SHEN Meihong(School of Acupuncture and Tuina/Regimen and Rehabilitation College,Nanjing University of Chinese Medicine,Nanjing,210023)
出处
《中医杂志》
CSCD
北大核心
2020年第15期1350-1356,共7页
Journal of Traditional Chinese Medicine
基金
国家自然科学基金(81774408,81973957)。
关键词
卵巢储备功能减退
针刺
动情周期
性激素
炎症因子
转录因子核因子E2相关因子2
血红素氧合酶1
diminished ovarian reserve
acupuncture
estrous cycle
sex hormones
inflammatory factors
transcription factor nuclear factor E2 related factor 2
heme oxygenase 1
