天津医药

天津医药 ›› 2022, Vol. 50 ›› Issue (6): 588-594.doi: 10.11958/20212673

• 实验研究 • 上一篇    下一篇

青蒿琥酯对多囊卵巢综合征模型大鼠卵巢组织形态的影响

韦奕，张淑芬，黄梦颖，马艳△   

  1. 南华大学附属第一医院妇产科（邮编421001）
  • 收稿日期:2021-12-02 修回日期:2022-02-23 出版日期:2022-06-15 发布日期:2023-12-20
  • 通讯作者: 韦奕 E-mail:weiyiyi957@163.com
  • 作者简介:统计见二修
  • 基金资助:
    湖南省卫生健康委科研立项课题（20201984）

The effects of artesunate on ovarian tissue morphology in rats with polycystic ovary syndrome

WEI Yi, ZHANG Shufen, HUANG Mengying, MA Yan△   

  1. Department of Obstetrics and Gynecology, the First Hospital Affiliated to Nanhua University, Hengyang 421001, China
  • Received:2021-12-02 Revised:2022-02-23 Published:2022-06-15 Online:2023-12-20

PDF (PC)

325

摘要： 摘要：目的　基于p38丝裂原活化蛋白激酶（p38 MAPK）/核因子-κB（NF-κB）信号通路探究青蒿琥酯（ART）对多囊卵巢综合征（PCOS）模型大鼠卵巢组织形态的影响。方法　对雌性SD大鼠皮下注射经过注射油剂稀释的脱氢表雄酮（1次/d），连续28 d，建立PCOS大鼠模型。造模成功的120只大鼠随机分为PCOS组、达英-35组、ART低剂量（25 mg/kg，ART-L）组、ART高剂量（50 mg/kg，ART-H）组、ART-H+p38 MAPK特异性激活剂茴香霉素（anisomycin）组，每组24只；另选取24只大鼠皮下注射等量生理盐水作为正常组。达英-35组、ART-L组、ART-H组、ART-H+anisomycin组大鼠连续28 d给予相应药物，正常组给予与PCOS组大鼠等量生理盐水，1次/d。吉姆萨染色观察大鼠动情周期阴道细胞形态；全自动生化分析仪检测大鼠空腹血糖（FPG）水平；酶联免疫吸附试验检测大鼠血清空腹胰岛素（FINS）、卵泡刺激素（FSH）、促黄体生成素（LH）、雌二醇（E2）、睾酮（T）、白细胞介素（IL）-18、肿瘤坏死因子（TNF）-α、IL-1β水平，计算胰岛素抵抗指数（HOMA-IR）；苏木素-伊红染色、透射电子显微镜分别观察卵巢组织病理情况和超微结构；Western blot法检测卵巢组织p38 MAPK/NF-κB通路蛋白表达。结果　与正常组比较，PCOS组大鼠动情周期紊乱，血清LH、T、FPG、FINS、HOMA-IR、IL-1β、TNF-α、IL-18水平、卵巢质量及卵泡数量、卵巢病理现象及损伤程度、磷酸化p38 MAPK（p-p38 MAPK）/p38 MAPK、磷酸化NF-κB p65（p-NF-κB p65）/NF-κB p65蛋白表达增加，FSH、E2水平降低（P＜0.05）；与PCOS组比较，达英-35组、ART-L组、ART-H组血清LH、T、FPG、FINS、HOMA-IR、IL-1β、TNF-α、IL-18水平、卵巢质量及卵泡数量、卵巢病理现象及损伤程度、p-p38 MAPK/p38 MAPK、p-NF-κB p65/NF-κB p65蛋白表达降低，FSH、E2水平增加（P＜0.05）；与达英-35组比较，ART-H组上述指标差异无统计学意义（P＞0.05）；与ART-H组比较，ART-H+anisomycin组血清LH、T、FPG、FINS、HOMA-IR、IL-1β、TNF-α、IL-18水平、卵巢质量及卵泡数量、卵巢病理现象及损伤程度、p-p38 MAPK/p38 MAPK、p-NF-κB p65/NF-κB p65表达增加，FSH、E2水平降低（P＜0.05）。结论　ART可能通过抑制p38 MAPK/NF-κB炎症通路活化，从而恢复PCOS大鼠性激素水平，改善卵巢组织形态及功能。

关键词: 多囊卵巢综合征；疾病模型, 动物；p38丝裂原活化蛋白激酶类；NF-κB；茴香霉素；青蒿琥酯；胰岛素抵抗

Abstract: Abstract: Objective　To explore the effect of artesunate (ART) on the ovarian tissue morphology of a rat model of polycystic ovary syndrome (PCOS) based on the p38 mitogen-activated protein kinase (p38 MAPK)/nuclear factor-κB (NF-κB) signaling pathway. Methods　The PCOS rat model was established by subcutaneous injection of dehydroepiandrosterone diluted with injection oil (once a day) for 28 days on SD female rats. A total of 120 rats with successful modeling were randomly divided into the PCOS group, the Diane-35 group, the ART low dose (25 mg/kg, ART-L) group, the ART high dose (50 mg/kg, ART-H) group and the ART-H+p38 MAPK specific activator (anisomycin) group, with 24 rats in each group. Another 24 rats were subcutaneously injected with the same amount of normal saline as the normal group. The rats in the Diane-35 group, the ART-L group, the ART-H group and the ART-H+anisomycin group were given corresponding drugs for 28 days, and the rats in the normal group and the PCOS group were given the same amount of normal saline, once a day. Giemsa staining was used to observe morphology of vaginal cells in ratestrous cycle. The fasting blood glucose (FPG) levels in rats were measured by automatic biochemical analyzer. Enzyme linked immunosorbent assay was used to detect the serum levels of fasting insulin (FINS), follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), testosterone (T), interleukin (IL)-18, tumor necrosis factor-α (TNF-α), and IL-1β. The insulin resistance index (HOMA-IR) was calculated. Ovarian histopathology and ultrastructure were observed by hematoxylin-eosin staining and transmission electron microscope, respectively. Western blot assay was used to detect the expression of p38 MAPK/NF-κB pathway protein in ovarian tissue. Results　Compared with the normal group, rats in the PCOS group had abnormal estrous cycle, decreased serum levels of LH, T, FPG, FINS, HOMA-IR, IL-1β, TNF-α and IL-18, decreased ovarian weight and number of follicles, heavier degrees of ovarian pathology and damage, increased expression levels of phosphorylated p38 MAPK (p-p38 MAPK)/p38 MAPK, phosphorylated NF-κB p65 (p-NF-κB p65)/NF-κB p65 protein, and reduced levels of FSH and E2 (P＜0.05). Compared with the PCOS group, the serum levels of LH, T, FPG, FINS, HOMA-IR, IL-1β, TNF-α and IL-18 were decreased, ovarian weight and number of follicles were decreased, the degrees of ovarian pathology and damage were aggravated, the expression levels of p-p38 MAPK/p38 MAPK, p-NF-κB p65/NF-κB p65 protein were reduced, and FSH and E2 levels were increased in the Diane-35 group, the ART-L group and the ART-H group (P＜0.05). Compared with the Diane-35 group, there were no significant differences in the above indicators in the ART-H group (P＞0.05). Compared with the ART-H group, the serum levels of LH, T, FPG, FINS, HOMA-IR, IL-1β, TNF-α and IL-18 were increased, ovarian weight and number of follicles were increased, the degrees of ovarian pathology and damage were aggravated, expression levels of p-p38 MAPK/p38 MAPK, p-NF-κB p65/NF-κB p65 protein were increased, and the levels of FSH and E2 were reduced in the ART-H+anisomycin group (P＜0.05). Conclusion　ART may restore the level of sex hormones and improve the morphology and function of ovarian tissue in PCOS rats by inhibiting the p38 MAPK/NF-κB inflammatory pathway.

Key words: polycystic ovary syndrome, disease models, animal, p38 mitogen-activated protein kinases, NF-kappa B, Anisomycin, Artesunate, insulin resistance