| 吴思怡,张观梅,王逸轩,丁彩飞,陈晓菲,雷蕾,周卢姗,方越,宋晓波,王晨晔.菟丝子-丹参药对通过线粒体功能途径抑制多囊卵巢综合征大鼠卵巢颗粒细胞凋亡研究[J].浙江中西医结合杂志,2025,35(11): |
| 菟丝子-丹参药对通过线粒体功能途径抑制多囊卵巢综合征大鼠卵巢颗粒细胞凋亡研究 |
| Cuscuta-Salvia inhibits polycystic ovary syndrome rat ovarian granulosa cells apoptosis through mitochondrial function pathway |
| 投稿时间：2025-02-02 修订日期：2025-06-13 |
| DOI： |
| 中文关键词: 多囊卵巢综合征 线粒体功能 颗粒细胞凋亡 菟丝子-丹参 卵泡发育 卵巢形态 |
| 英文关键词:polycystic ovary syndrome mitochondrial function granulosa cells apoptosis Cuscuta-Salvia follicular development ovarian morphology |
| 基金项目:]浙江省中医药科技计划项目(2021ZB210、2024ZF109)；浙江中医药大学科研项目(2021FSYYZY30)；浙大预研项目(2022009)；中医药治疗妇科内分泌与代谢疾病创新团队(2022SJCXTD) |
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| 中文摘要: |
| 目的 探究菟丝子-丹参药对对多囊卵巢综合征（PCOS）大鼠卵巢颗粒细胞凋亡及线粒体功能的影响。方法 40只雌性SD大鼠随机分为对照组、模型组、二甲双胍组和药对组（菟丝子-丹参组），每组10只。除对照组外均使用来曲唑灌胃法造模21天，其中二甲双胍组予二甲双胍药物溶液50mg/kg灌胃、药对组予菟丝子-丹参提取物溶液0.1ml/10g灌胃，对照组、模型组予以等量生理盐水灌胃，持续干预21天。记录大鼠体重、卵巢质量，检测空腹血糖；苏木素-伊红（HE）染色观察大鼠卵巢组织形态学变化；TUNEL染色观察大鼠原代卵巢颗粒细胞凋亡情况。免疫组织化学( IHC )染色和蛋白免疫印迹法（WB）检测大鼠卵巢组织BAX、BCL-2蛋白表达。mPTP法检测线粒体通透性转换孔开放程度；ATP生物发光法检测细胞能量代谢水平；COXIV法检测细胞色素c氧化酶的功能性活性；JC-1法检测线粒体膜电位变化。结果 与对照组比较，模型组大鼠的卵巢质量[ （104.33±1.53）mg比 （159.67±6.03）mg，P<0.05]、大鼠体质量[ （290.33±4.51）mg比 （327.00±33.29）mg，P＞0.05]及空腹血糖水平[ （4.43±0.25）mmol/L 比 （5.87±0.55mmol/L，P<0.05]均升高。通过HE染色及TUNEL染色提示，与模型组相比，药对组大鼠的卵巢组织形态改善、卵泡发育较好、动情周期紊乱、卵巢颗粒细胞凋亡数减少[ （0.19±0.03）%比 （0.16±0.03）%，P＞0.05]。免疫组织化学染色显示，与模型组相比，药对组大鼠的BAX蛋白表达[ （0.28±0.09）比 （0.21±0.07），P＞0.05]显著上升，BCL-2蛋白表达 [ （0.13±0.4）比 （0.18±0.07），P＞0.05]显著下降。Western blotting也显示，与模型组相比，药对组大鼠的BAX/GADPH[ （1.10±0.03）比 （0.86±0.05），P＜0.05]显著上升，BCL-2/GADPH[ （0.90±0.07）比 （1.50±0.09），P＜0.05]显著下降。线粒体功能各项实验中显示，与模型组相比，药对组大鼠的mPTP[ （35.07±7.70）比 （37.28±9.18），P＞0.05]、ATP[ （0.935±0.014）μm比 （1.448±0.072）μm，P＜0.05]、COXIV[ （0.0117±0.0028）μmol比 （0.0232±0.0022）μmol，P＜0.05]、JC-1水平[ （2.819±1.586）%比 （5.338±1.011）%，P＞0.05]均显著升高。
结论 菟丝子-丹参药对可通过恢复线粒体功能、抑制卵巢颗粒细胞凋亡，促进卵泡发育情况，起到治疗PCOS的作用。 |
| 英文摘要: |
| Abstract: Objective To explore the effects of Cuscuta-Salvia groups on the apoptosis of ovarian granulosa cells and mitochondrial function in rats with polycystic ovary syndrome (PCOS).
Methods Forty female SD rats were randomly divided into the control group, the model group, the metformin group and the drug pair group (Cuscuta chinensis - Salvia miltiorrhiza group), with 10 rats in each group. Except for the control group, all others were modeled by the letrozole gavage method for 21 days. Among them, the metformin group was given a 50mg/kg metformin drug solution by gavage, the drug pair group was given a 0.1ml/10g Cuscuta chinensis - Salvia miltiorrhiza extract solution by gavage, and the control group and the model group were given the same amount of normal saline by gavage. The intervention lasted for 21 days. Record the body weight and ovarian mass of rats and detect fasting blood glucose; Hematoxylin-eosin (HE) staining was used to observe the morphological changes of ovarian tissue in rats; TUNEL staining was used to observe the apoptosis of primary ovarian granulosa cells in rats. The protein expressions of BAX and BCL-2 in rat ovarian tissues were detected by immunohistochemical (IHC) staining and Western blotting (WB). The opening degree of mitochondrial permeability transition pores was detected by the mPTP method; The energy metabolism level of cells was detected by ATP-bioluminescence assay; The functional activity of cytochrome c oxidase was detected by the COXIV method; The changes of mitochondrial membrane potential were detected by JC-1 method.Results Compared with the control group, the ovarian mass of rats in the model group [(104.33±1.53) mg vs. (159.67±6.03) mg, P<0.05] and the body weight of rats [(290.33±4.51) mg vs. (327.00±33.29) mg] Both the fasting blood glucose level [(4.43±0.25) mmol/L vs. (5.87±0.55mmol/L, P<0.05] increased. HE staining and TUNEL staining indicated that, compared with the model group, the ovarian tissue morphology of rats in the drug pair group was improved, follicular development was better, estrous cycle was disordered, and the number of apoptotic ovarian granulosa cells was reduced [(0.19±0.03) % vs. (0.16±0.03) %, P > 0.05]. Immunohistochemical staining showed that compared with the model group, the expression of BAX protein in the drug pair group of rats [(0.28±0.09) vs. (0.21±0.07), P > 0.05] increased significantly, and the expression of BCL-2 protein [(0.13±0.4) vs. (0.18±0.07), P > 0.05] decreased significantly. Western blotting also showed that compared with the model group, the BAX/GADPH of rats in the drug pair group [(1.10±0.03) vs. (0.86±0.05), P < 0.05] increased significantly, and BCL-2/GADPH[(0.90±0.07) vs. (1.50±0.09), [P < 0.05] Decreased significantly. In various experiments of mitochondrial function, it was shown that compared with the model group, the mPTP[(35.07±7.70) vs. (37.28±9.18), P > 0.05] and ATP[(0.935±0.014) μm vs. (1.448±0.072) μm] of rats in the drug pair group P < 0.05], COXIV[(0.0117±0.0028) μmol vs. (0.0232±0.0022) μmol, P < 0.05], and JC-1 levels [(2.819±1.586) % vs. (5.338±1.011) %, P > 0.05] were all significantly increased.Conclusion Cuscuta-Salvia can play a therapeutic role in PCOS by restoring mitochondrial function, inhibiting the apoptosis of ovarian granulosa cells and promoting the development of follicles. |
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