基于线粒体裂变探讨艾灸对早发性卵巢功能不全小鼠卵巢功能的保护作用

罗佳; 尹尧丽; 沈梅红; 房延苏; 朱晨歌; 范彦良

doi:10.13703/j.0255-2930.20241126-k0006

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基于线粒体裂变探讨艾灸对早发性卵巢功能不全小鼠卵巢功能的保护作用

机制探讨 | 更新时间：2026-03-16

- 基于线粒体裂变探讨艾灸对早发性卵巢功能不全小鼠卵巢功能的保护作用
- Protective effect of moxibustion on ovarian function in mice with premature ovarian insufficiency based on mitochondrial fission
- 中国针灸 2026年46卷第3期页码：411-420
- 作者机构：
  
  南京中医药大学针灸推拿学院·养生康复学院，江苏南京　210023
  湖南医药学院，怀化　418000
- 作者简介：
  
  罗佳，南京中医药大学硕士研究生。E-mail：lj07182022@163.com
  ✉沈梅红，教授。E-mail：mhshen@njucm.edu.cn
- 基金信息：
  
  国家自然科学基金资助项目(82374599);江苏省中医药领军人才培养项目(SLJ0316);江苏省研究生科研创新计划项目(KYCX24_2189)
- DOI：10.13703/j.0255-2930.20241126-k0006
  中图分类号：
- 收稿：2024-11-26，
  
  网络首发：2025-04-29，
  
  纸质出版：2026-03-12
- 稿件说明：
移动端阅览
罗佳, 尹尧丽, 沈梅红, 等. 基于线粒体裂变探讨艾灸对早发性卵巢功能不全小鼠卵巢功能的保护作用[J]. 中国针灸, 2026,46(3):411-420.

LUO Jia, YIN Yaoli, SHEN Meihong, et al. Protective effect of moxibustion on ovarian function in mice with premature ovarian insufficiency based on mitochondrial fission[J]. Chinese Acupuncture & Moxibustion, 2026, 46(3): 411-420.
罗佳, 尹尧丽, 沈梅红, 等. 基于线粒体裂变探讨艾灸对早发性卵巢功能不全小鼠卵巢功能的保护作用[J]. 中国针灸, 2026,46(3):411-420. DOI： 10.13703/j.0255-2930.20241126-k0006.

LUO Jia, YIN Yaoli, SHEN Meihong, et al. Protective effect of moxibustion on ovarian function in mice with premature ovarian insufficiency based on mitochondrial fission[J]. Chinese Acupuncture & Moxibustion, 2026, 46(3): 411-420. DOI： 10.13703/j.0255-2930.20241126-k0006.

摘要

目的：

利用线粒体裂变激活剂，探讨艾灸对早发性卵巢功能不全（POI）小鼠卵巢功能的保护作用是否与卵巢颗粒细胞线粒体动力学、功能有关。

方法：

将80只动情周期规律的雌性ICR小鼠随机分为空白组、雷公藤多苷组（TG组）、艾灸组、羰基氰化物3-氯苯腙+艾灸组（CCCP+艾灸组），每组20只。除空白组外，其余组通过连续14 d灌胃TG混悬液制备POI模型。艾灸组于TG灌胃1 h后交替艾灸双侧“肾俞”或“中脘”“关元”穴，每日1次，干预14 d。CCCP+艾灸组在TG灌胃后腹腔注射CCCP（2.9 mg·kg

-1

·d

-1

），与艾灸组同样施灸。记录小鼠动情周期，计算卵巢指数；HE染色法观察小鼠卵巢组织形态，并进行卵泡计数；ELISA法检测小鼠血清性激素[卵泡刺激素（FSH）、雌二醇（E

）、抗缪勒管激素（AMH）

]

含量；免疫组化染色法和实时荧光定量PCR法检测卵巢线粒体动力学相关因子[动力蛋白相关蛋白1（Drp1）、线粒体裂变因子（Mff）、视神经萎缩1（Opa1）、线粒体融合蛋白 1（Mfn1）

]

的蛋白和mRNA表达；荧光素酶法和DCFH-DA荧光探针法分别检测颗粒细胞三磷酸腺苷（ATP）浓度和活性氧（ROS）荧光强度。

结果：

与空白组比较，TG组小鼠动情周期紊乱率升高（

0.01），卵巢体积较小，卵巢指数降低（

0.01），原始卵泡、生长卵泡、窦卵泡数量减少（

0.01），闭锁卵泡数量增多（

0.01），卵巢颗粒细胞排列疏松紊乱；血清FSH含量升高（

0.01），血清AMH、E

含量降低（

0.01）；卵巢Drp1、Mff蛋白颗粒细胞染色深，卵巢Opa1、Mfn1蛋白染色浅，卵巢Drp1、Mff平均吸光度及卵巢颗粒细胞Drp1、Mff mRNA表达升高（

0.01），Opa1、Mfn1平均吸光度及卵巢颗粒细胞Opa1、Mfn1 mRNA表达降低（

0.01），卵巢颗粒细胞ATP浓度降低（

0.01），ROS荧光强度升高（

0.01）。与TG组比较，艾灸组小鼠动情周期紊乱率降低（

0.01），卵巢体积较大，卵巢指数升高（

0.01），原始卵泡、生长卵泡、窦卵泡数量增加（

0.01），闭锁卵泡数量减少（

0.01），卵巢颗粒细胞排列较紧密有序；血清FSH含量降低（

0.01），血清AMH、E

含量升高（

0.01）；卵巢Opa1、Mfn1蛋白染色深，卵巢Drp1、Mff蛋白染色浅，卵巢Opa1、Mfn1平均吸光度及卵巢颗粒细胞Opa1、Mfn1 mRNA表达升高（

0.01，

0.05），卵巢Drp1、Mff平均吸光度及卵巢颗粒细胞Drp1、Mff mRNA表达降低（

0.05，

0.01），卵巢颗粒细胞ATP浓度升高（

0.01），ROS荧光强度降低（

0.01）。与艾灸组比较，CCCP+艾灸组小鼠动情周期紊乱率升高（

0.05），卵巢指数降低（

0.05），原始卵泡、生长卵泡、窦卵泡数量减少（

0.01，

0.05），闭锁卵泡数量增加（

0.01），卵巢颗粒细胞排列较疏松紊乱；血清FSH含量升高（

0.01），血清E

、AMH含量降低（

0.05）；卵巢Drp1、Mff蛋白染色深，卵巢Opa1、Mfn1蛋白染色浅，卵巢Drp1、Mff平均吸光度及卵巢颗粒细胞Drp1、Mff mRNA表达升高（

0.05），卵巢Opa1、Mfn1平均吸光度及卵巢颗粒细胞Opa1、Mfn1 mRNA表达降低（

0.05，

0.01），卵巢颗粒细胞ATP浓度降低（

0.01），ROS荧光强度升高（

0.01）。

结论：

艾灸可能通过抑制线粒体裂变，增加线粒体融合从而改善受损的线粒体功能以减缓TG所致的卵巢损伤，且此作用可被CCCP阻断。

Abstract

Objective

To investigate whether the protective effect of moxibustion on ovarian function in mice with premature ovarian insufficiency (POI) was associated with mitochondrial dynamics and function in ovarian granulosa cells

using a mitochondrial fission activator.

Methods

Eighty female ICR mice with regular estrous cycles were randomly divided into a blank group

a tripterygium glycosides group (TG group)

a moxibustion group

and a carbonyl cyanide m-chlorophenyl hydrazone plus moxibustion group (CCCP+moxibustion group)

with 20 mice in each group. Except for the blank group

POI model was established in the remained groups by intragastric administration of TG suspension for 14 consecutive days. In the moxibustion group

moxibustion was applied at bilateral "Shenshu" (BL23) or alternately at "Zhongwan" (CV12) and "Guanyuan" (CV4) 1 hour after TG administration

once every day for 14 days. The CCCP+moxibustion group received intraperitoneal injection of CCCP (2.9 mg·kg

-1

·d

-1

) after TG administration

followed by the same moxibustion protocol as the moxibustion group. Estrous cycles were recorded

and the ovarian index was calculated. Ovarian morphology was observed with HE staining

and follicle counts were conducted. Serum levels of follicle-stimulating hormone (FSH)

estradiol (E

)

and anti-Müllerian hormone (AMH) were measured using ELISA. Immunohistochemistry and real-time quantitative PCR were used to detect the protein and mRNA expression of mitochondrial dynamics-related factors (dynamin-related protein 1 [Drp1

]

mitochondrial fission factor [Mff

]

optic atrophy 1 [Opa1

]

and mitofusin 1 [Mfn1

]

) in the ovaries

respectively. ATP concentration in granulosa cells was determined by luciferase assay

and reactive oxygen species (ROS) fluorescence intensity was detected using DCFH-DA fluorescent probe method.

Results

Compared with the blank group

the mice in the TG group showed a significantly higher rate of estrous cycle disorder (

0.01)

smaller ovarian volume

lower ovarian index (

0.01)

reduced numbers of primordial

growing

and antral follicles (

0.01)

increased number of atretic follicles (

0.01)

and loose and disordered arrangement of granulosa cells; serum FSH level was elevated (

0.01)

while serum AMH and E

levels were reduced (

0.01). Drp1 and Mff protein staining in ovarian granulosa cells was intensified

while Opa1 and Mfn1 protein staining was lighter. The average optical densities of Drp1 and Mff and the mRNA expression of Drp1 and Mff were significantly increased (

0.01)

while the average optical densities of Opa1 and Mfn1 and the mRNA expression of Opa1 and Mfn1 were significantly decreased (

0.01). ATP concentration was reduced (

0.01)

and ROS fluorescence intensity was decreased (

0.01). Compared with the TG group

the mice in the moxibustion group exhibited lower rate of estrous cycle disorder (

0.01)

larger ovarian volume

higher ovarian index (

0.01)

increased numbers of primordial

growing

and antral follicles (

0.01)

reduced number of atretic follicles (

0.01)

and a more compact and orderly arrangement of granulosa cells; serum FSH level was decreased (

0.01)

while serum AMH and E

levels were increased (

0.01). Opa1 and Mfn1 protein staining was deepened

Drp1 and Mff protein staining was lightened

average optical densities and mRNA expression of Opa1 and Mfn1 were increased (

0.01

0.05)

and those of Drp1 and Mff were decreased (

0.05

0.01). ATP concentration was increased (

0.01)

and ROS fluorescence intensity was decreased (

0.01). Compared with the moxibustion group

the mice in the CCCP+moxibustion group showed an increased estrous cycle disorder rate (

0.05) and a decreased

ovarian index (

0.05). The numbers of primordial

growing

and antral follicles were reduced (

0.01

0.05)

while the number of atretic follicles was increased (

0.01). The arrangement of ovarian granulosa cells became looser and more disordered. Serum FSH level was elevated (

0.01)

while serum E

and AMH levels were decreased (

0.05). In the ovaries of mice in the CCCP+moxibustion group

Drp1 and Mff protein staining was intensified

whereas Opa1 and Mfn1 protein staining was lighter. The average optical densities of Drp1 and Mff

as well as the mRNA expression levels of Drp1 and Mff in granulosa cells

were increased (

0.05)

while the average optical densities of Opa1 and Mfn1 and the mRNA expression levels of Opa1 and Mfn1 in granulosa cells were decreased (

0.05

0.01). Additionally

the ATP concentration in ovarian granulosa cells was reduced (

0.01)

and the ROS fluorescence intensity was elevated (

0.01).

Conclusion

Moxibustion may alleviate TG-induced ovarian damage by inhibiting mitochondrial fission and promoting mitochondrial fusion

thereby improving impaired mitochondrial function; this protective effect can be blocked by CCCP.

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