Shuai Ling, She Jiajie, Diao Ruiying, Zhao Huihui, Liu Xuemin, Hu Qingyu, Li Dongdong, Su Danna, Ye Xiaofeng, Guo Yan, Zhong Mei, Wang Liping
Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical University, Guangzhou, China.
The First Affiliated Hospital of Shenzhen University, Reproductive Medicine Centre, Shenzhen Second People's Hospital, Shenzhen, China.
Am J Reprod Immunol. 2023 Apr;89(4):e13686. doi: 10.1111/aji.13686. Epub 2023 Feb 19.
The role of hydroxychloroquine (HCQ) in premature ovarian insufficiency (POI) remains unclear. The purpose of this study was to evaluate the effect of HCQ on ovarian function in mice with POI and to clarify its potential mechanisms.
POI was induced in mice by injection with zona pellucida 3 peptide (pZP3), and HCQ was administered intragastrically. Stages of the estrous cycle were determined using vaginal cytology. The ovarian structure was observed under a microscope after hematoxylin-eosin staining. The levels of serum hormones and anti-ZP antibody (aZPAb) were measured using enzyme-linked immunosorbent assay (ELISA). The expression levels of CD4, CD45, and ZP2, ZP3 were determined using immunofluorescence and immunohistochemistry, respectively. The T regulatory (Treg)/ T helper 17 (Th17) cell ratio was analyzed using flow cytometry analysis. Western blotting was performed to assess the expression levels of proteins, transcription factors and cytokines.
Administration of HCQ to mice with POI greatly restored their estrus cycle. In the treatment group compared to the POI group, estradiol (E ) levels were higher, and follicle stimulating hormone (FSH) levels were lower. In addition, following pZP3, HCQ treatment increased ZP2 and ZP3 expression. Additionally, by inhibiting the activation of the TLR7 signaling pathway, HCQ attenuated the infiltration of inflammatory cells and prevented the activated naive CD4 T cells from developing into Th17 cells.
Our findings showed that HCQ effectively restored ovarian function by altering the Treg/Th17 cell ratio in mice with POI, indicating that HCQ maybe a promising therapeutic method for patients with POI.
羟氯喹（HCQ）在卵巢早衰（POI）中的作用尚不清楚。本研究旨在评估HCQ对POI小鼠卵巢功能的影响，并阐明其潜在机制。
通过注射透明带3肽（pZP3）诱导小鼠发生POI，并对其进行HCQ灌胃给药。采用阴道细胞学检查确定发情周期阶段。苏木精-伊红染色后在显微镜下观察卵巢结构。采用酶联免疫吸附测定（ELISA）检测血清激素和抗ZP抗体（aZPAb）水平。分别采用免疫荧光和免疫组织化学法测定CD4、CD45以及ZP2、ZP3的表达水平。采用流式细胞术分析调节性T（Treg）细胞/辅助性T细胞17（Th17）细胞比例。进行蛋白质印迹法以评估蛋白质、转录因子和细胞因子的表达水平。
对POI小鼠给予HCQ可极大恢复其发情周期。与POI组相比，治疗组的雌二醇（E）水平较高，卵泡刺激素（FSH）水平较低。此外，在给予pZP3后，HCQ治疗可增加ZP2和ZP3的表达。此外，通过抑制TLR7信号通路的激活，HCQ可减轻炎性细胞浸润，并阻止活化的初始CD4 T细胞分化为Th17细胞。
我们的研究结果表明，HCQ通过改变POI小鼠的Treg/Th17细胞比例有效恢复了卵巢功能，这表明HCQ可能是一种有前景的POI患者治疗方法。
