Wang Qian, Lu Ge, Xie Zi-Jing, Li Hong-Xiao, Shen Mei-Hong
College of Acupuncture-Moxibustion and Tuina, Nanjing University of Chinese Medicine, Nanjing 210023, Jiangsu Province, China.
Zhongguo Zhen Jiu. 2021 Jan 12;41(1):53-8. doi: 10.13703/j.0255-2930.20191128-k0005.
To observe the effect of moxibustion on Nrf2/HO-1 signaling pathway in rats with diminished ovarian reserve (DOR), and to explore the protective mechanism of moxibustion on ovarian reserve function.
Forty SD rats were randomly divided into a blank group, a model group, a moxibustion group and a hormone group, 10 rats in each group. The rats in the model group, moxibustion group and hormone group were treated with intragastric administration of tripterysium glycosides turbid liquid to prepare DOR model. The rats in the blank group were treated with intragastric administration of sodium chloride solution with the same volume, once a day for 14 days. The rats in the hormone group were treated with hormone sequential therapy for 14 days from the day of modeling; the rats in the moxibustion group were treated with moxibustion at bilateral "Shenshu" (BL 23) or "Guanyuan" (CV 4) and "Zhongwan" (CV 12) from the day of modeling, and the two groups acupoints were alternated every other day, 10 min each time, for 14 consecutive days. The estrus cycle was observed every day by vaginal exfoliated cell smear, and the estrus cycle disorder rate in each group was calculated. After the intervention, the HE staining was used to observe the histological morphology of ovaries; ELISA was used to detect the contents of follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E), anti Mullerian hormone (AMH), superoxide dismutase (SOD) and malondialdehyde (MDA); the protein levels of Nrf2 and HO-1 in ovarian tissue were detected by immunohistochemistry; real-time PCR (TaqMan probe method) was used to detect the expression of Nrf2 and HO-1 mRNA.
Compared with the blank group, the rate of estrus cycle disorder in the model group was increased (<0.01); compared with the model group, the rate of estrus cycle disorder in the moxibustion group and hormone group was decreased (<0.01). Compared with the blank group, the serum contents of FSH, LH and MDA in the model group were increased (<0.01), and the serum contents of E, AMH and SOD were decreased (<0.01). Compared with the model group, the serum contents of FSH, LH and MDA in the moxibustion group and hormone group were decreased (<0.01, <0.05), and the serum contents of E, AMH and SOD were increased (<0.01). Compared with the blank group, the protein and mRNA expression of Nrf2 and HO-1 in the model group were decreased (<0.01); compared with the model group, the protein and mRNA expressions of Nrf2 and HO-1 in the moxibustion group and hormone group were increased (<0.01).
Moxibustion could reduce the rate of estrus cycle disorder, improve the level of serum sex hormones and antioxidant stress in DOR rats, and the mechanism may be related to the regulation of Nrf2/HO-1 signaling pathway.
观察艾灸对卵巢储备功能下降（DOR）大鼠核因子E2相关因子2（Nrf2）/血红素加氧酶-1（HO-1）信号通路的影响，探讨艾灸对卵巢储备功能的保护机制。
将40只SD大鼠随机分为空白组、模型组、艾灸组和激素组，每组10只。模型组、艾灸组和激素组大鼠采用灌胃雷公藤多苷混悬液制备DOR模型。空白组大鼠灌胃等体积氯化钠溶液，每日1次，连续14天。激素组大鼠自造模之日起采用激素序贯疗法治疗14天；艾灸组大鼠自造模之日起在双侧“肾俞”（BL 23）或“关元”（CV 4）及“中脘”（CV 12）进行艾灸，两组穴位隔日交替，每次10分钟，连续14天。每日通过阴道脱落细胞涂片观察动情周期，计算每组动情周期紊乱率。干预后，采用苏木精-伊红（HE）染色观察卵巢组织形态学；采用酶联免疫吸附测定（ELISA）法检测卵泡刺激素（FSH）、黄体生成素（LH）、雌二醇（E）、抗苗勒管激素（AMH）、超氧化物歧化酶（SOD）和丙二醛（MDA）含量；采用免疫组织化学法检测卵巢组织中Nrf2和HO-1蛋白水平；采用实时荧光定量聚合酶链反应（TaqMan探针法）检测Nrf2和HO-1 mRNA表达。
与空白组比较，模型组动情周期紊乱率升高（<0.01）；与模型组比较，艾灸组和激素组动情周期紊乱率降低（<0.01）。与空白组比较，模型组血清FSH、LH和MDA含量升高（<0.01），血清E、AMH和SOD含量降低（<0.01）。与模型组比较，艾灸组和激素组血清FSH、LH和MDA含量降低（<0.01，<0.05），血清E、AMH和SOD含量升高（<0.01）。与空白组比较，模型组Nrf2和HO-1蛋白及mRNA表达降低（<0.01）；与模型组比较，艾灸组和激素组Nrf2和HO-1蛋白及mRNA表达升高（<0.01）。
艾灸可降低DOR大鼠动情周期紊乱率，提高血清性激素水平及抗氧化应激能力，其机制可能与调控Nrf2/HO-1信号通路有关。
