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目的:探讨苦瓜总皂苷对卵巢功能是否有有益作用及其可能的作用机制,以促进我国妇科学的发展。方法:选取雌性SD大鼠60只,随机分为正常对照组,模型对照组,苦瓜总皂苷低(10 mg·kg~(-1)·d~(-1))、中(20 mg·kg~(-1)·d~(-1))和高剂量(40 mg·kg~(-1)·d~(-1))治疗组以及阳性药物逍遥丸干预组,大鼠在适应性喂养2周后,除正常对照组,其余5组给予颈背部皮下1.25 mg丙酸睾丸酮一次性注射造模,待大鼠阴道开口后,检测其阴道上皮是否有持续角化来判定排卵障碍模型大鼠是否成功。造模成功后给予相应药物治疗,正常对照组和模型对照组只给予对应量生理盐水灌胃;治疗6周后处死大鼠,收集血清和卵巢组织,测定其血清中胆红素、总抗氧化能力(T-AOC)和丙二醛(MDA);同时免疫组化、Western以及半定量PCR观察卵巢组织Caspase-3蛋白的表达特点。结果:与正常组相比,模型组总胆红素、直接胆红素以及丙二醛明显升高(P<0.05),总抗氧化能力明显下降(P<0.05),苦瓜总皂苷治疗后,相较于模型组,3个治疗组以及逍遥丸干预组总胆红素、直接胆红素以及丙二醛明显下降(P<0.05),其中中剂量和高剂量组和正常对照组一致;此外苦瓜总皂苷治疗后,卵巢颗粒细胞免疫组化、Western blot以及半定量PCR检测Caspase-3结果表明,模型组卵巢细胞Caspase-3表达量较多,而3个治疗组均可以明显的减少卵巢细胞Caspase-3蛋白表达,且高剂量组效果优于低剂量组,中高剂量组优于逍遥丸治疗组。结论:苦瓜总皂苷能改善排卵障碍模型大鼠卵巢的氧化损伤和细胞凋亡,推测其是通过此途径来改善卵巢功能,同时其效果优于目前临床中常用的逍遥丸。
Objective: To investigate whether the bitter melon total saponins have beneficial effects on ovarian function and its possible mechanism to promote the development of gynecology in our country. Methods: Sixty female Sprague-Dawley rats were randomly divided into normal control group, model control group, low bitter melon (10 mg · kg -1 · d -1), medium (20 mg · kg -1 (-1) · d -1 and high dose (40 mg · kg -1 · d -1) groups, as well as the intervention group of Xiaoyao Wan, Two weeks later, except for the normal control group, the other five groups were injected subcutaneously with 1.25 mg of testosterone propionate on the back of the neck. After the vagina was opened, the vaginal epithelium was examined for persistent keratinization to determine the ovulation disorder model rats whether succeed. After the model was established successfully, the corresponding medicine was given. The normal control group and the model control group were only given the corresponding amount of normal saline. After 6 weeks of treatment, the rats were sacrificed and the serum and ovarian tissues were collected. The serum bilirubin, total antioxidant (T-AOC) and malondialdehyde (MDA). Meanwhile, the expression of Caspase-3 protein in ovarian tissue was detected by immunohistochemistry, Western and semi-quantitative PCR. Results: Compared with normal group, total bilirubin, direct bilirubin and malondialdehyde in model group were significantly increased (P <0.05), total antioxidant capacity decreased significantly (P <0.05) Compared with the model group, the levels of total bilirubin, direct bilirubin, and malondialdehyde in the three treatment groups and the Xiaoyao Pill intervention group were significantly decreased (P <0.05), and the middle and high dose groups were the same as the normal control group; in addition Caspase-3 was detected by immunohistochemistry of ovarian granulosa cells, Western blot and semiquantitative PCR after treatment with total saponins of Momordia charantia. The results showed that the expression of Caspase-3 in the model group was higher than that in the untreated group, and all the three treatment groups could significantly reduce the ovarian cells Caspase-3 protein expression, and high-dose group is better than the low-dose group, medium-high dose group is better than the Xiaoyao Pill group. Conclusion: The total saponin of Momordica charantia can improve ovarian oxidative damage and apoptosis in ovariectomized rats. It is presumed that this method can improve ovarian function and its effect is superior to the commonly used Xiaoyao Wan in clinic.