| 李晓珂,韩亚光,韩延华,王宝龙,梁 霄,杨 婧.黄芩苷对多囊卵巢综合征卵巢颗粒细胞增殖和凋亡的影响及机制[J].,2024,(21):4007-4013 |
| 黄芩苷对多囊卵巢综合征卵巢颗粒细胞增殖和凋亡的影响及机制 |
| Effects of Baicalin on the Proliferation and Apoptosis of Ovarian Granulosa Cells in Polycystic Ovary Syndrome |
| 投稿时间：2024-06-04 修订日期：2024-06-30 |
| DOI：10.13241/j.cnki.pmb.2024.21.002 |
| 中文关键词: 黄芩苷 多囊卵巢综合征 颗粒细胞 细胞增殖 细胞凋亡 |
| 英文关键词: Baicalin Polycystic ovary syndrome Granulosa cell Cell multiplication Apoptosis |
| 基金项目:国家自然科学基金项目（81603418）；黑龙江省自然科学基金项目（LH2023H060） |
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| 中文摘要: |
| 摘要 目的：探讨黄芩苷（baicalin, BAI）对多囊卵巢综合征（polycystic ovary syndrome, PCOS）大鼠卵巢颗粒细胞增殖和凋亡的影响及机制。方法：将48只雌性SD大鼠随机分为对照组、PCOS组、BAI低、中、高剂量组及阳性对照组，每组8只。通过皮下注射（dehydroepiandros-terone, DHEA）建立PCOS模型，BAI低、中、高剂量组大鼠分别给予黄芩苷（12.5、25、50 mg/kg）灌胃；阳性对照组大鼠给予（10 mg/kg维替泊芬）腹腔注射；对照组和PCOS组大鼠给予等体积0.9% NaCl溶液代替。每日1次，持续4周。采用（enzyme-linked immunosorbent assay, ELISA）法检测大鼠血清中性激素水平；HE染色观察大鼠卵巢病理学变化。造模成功后，提取卵巢颗粒细胞，3-（4,5-二甲基噻唑-2）-2，5-二苯基四氮唑溴盐法（3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide, MTT）法检测卵巢颗粒细胞增殖情况；Annexin V-FITC/PI双染色法和TUNEL染色法检测凋亡情况；蛋白质印记法检测凋亡相关蛋白Bcl-2、Bax，P13K/AKT通路相关蛋白P13K、AKT、p-P13K、p-AKT的相对表达。结果：提取的卵巢颗粒细胞呈成纤维细胞样、贴壁良好，特异性蛋白FSHR均呈阳性表达。与对照组相比，PCOS组大鼠卵巢结构紊乱，出血、充血及囊状卵泡数量增多，白膜增厚；血清睾酮（testosterone, T）、黄体生成素（luteinizing hormone, LH）水平、卵巢颗粒细胞凋亡率及Bax蛋白表达显著升高（P<0.05），而血清雌二醇（estradiolm, E2）、卵泡刺激素（follicle stimulating hormone, FSH）水平、卵巢颗粒细胞生存活力、Bcl-2、p-P13K、及p-AKT蛋白表达显著下降（P<0.05）；与PCOS组相比，BAI低、中、高剂量组多囊特征恢复明显、囊肿显著减少、卵巢结构及颗粒细胞层排列完整；血清E2、FSH水平、卵巢颗粒细胞生存活力、Bcl-2、p-P13K、及p-AKT蛋白表达呈剂量依赖性显著上调（P<0.05），而血清T、LH水平、卵巢颗粒细胞凋亡率及Bax蛋白表达呈剂量依赖性显著下调（P<0.05）。结论：黄芩苷可能通过激活P13K/AKT信号通路促进卵巢颗粒细胞增殖并抑制其凋亡，进而减轻PCOS症状。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effect of baicalin (BAI) on the proliferation and apoptosis of ovarian granulosa cells in polycystic ovary syndrome (PCOS) and its mechanisms. Methods: Forty-eight female SD rats were randomly divided into the control group, PCOS group, BAI low, middle and high dose groups and positive control group, with 8 rats in each group. PCOS model was established by subcutaneous injection of DHEA, and rats in the low, medium, and high dose groups of BAI were given baicalin (12.5, 25, and 50 mg/kg) by gavage. The Positive control group was given intraperitoneal injection (10 mg/kg) of Vitipofen; The control group and PCOS group were given equal volume 0.9% NaCl solution instead. Once a day for 4 weeks. The serum sex hormone levels were detected by enzyme-linked immunosorbant assay (ELISA). The pathological changes of ovary were observed by HE staining. After successful modeling, ovarian granulosa cells were extracted, and their proliferation was detected by MTT assay. Apoptosis was detected by AnnexinV-FITC/PI double staining and TUNEL staining. The relative expressions of BCL2 and BAX, P13K/AKT pathway related proteins P13K, AKT, p-P13K and p-AKT were detected by western blot. Results: The extracted ovarian granulosa cells are fibroblast-like and adhered well, with positive expression of specific proteins FSHR. Compared with the control group, the ovarian structure of rats in the PCOS group is disrupted, with an increase in bleeding, congestion, and the number of cystic follicles, as well as thickening of the white membrane. The levels of serum testosterone (T) and luteinizing hormone (LH), the apoptosis rate of ovarian granulosa cells and the expression of Bax protein are significantly increased (P<0.05), while the levels of Estradiolm (E2) and follicle stimulating hormone (FSH), the viability of ovarian granulosa cells, the expression of Bcl-2, p-P13K, and p-AKT protein are significantly decreased (P<0.05). Compared with PCOS group, BAI groups have obvious recovery of polycystic features, significant reduction in cysts, and complete arrangement of ovarian structure and granular cell layer. The levels of E2 and FSH in serum, the viability of ovarian granulosa cells, the expressions of Bcl-2, p-P13K and p-AKT protein are significantly increased in a dose-dependent manner (P<0.05), while the levels of T and LH in serum, the apoptosis rate of ovarian granulosa cells and the expression of Bax protein are significantly decreased in a dose-dependent manner (P<0.05). Conclusion: Baicalin may promote the proliferation and inhibit the apoptosis of ovarian granulosa cell, which may alleviate PCOS by activating the P13K/AKT signaling pathway. |
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