壮医滋勒调经汤干预卵巢储备功能低下大鼠模型的实验研究

发布时间：2018-04-30 00:34

本文选题：滋勒调经汤 + 卵巢储备功能低下　；参考：《广西中医药大学》2017年硕士论文

【摘要】：目的:通过建立卵巢储备功能低下(diminished ovarian reserve,DOR)大鼠模型,以滋勒调经汤进行干预,通过观察卵巢形态、大鼠性激素和OSM、M-CSF细胞因子等相关指标,初步探讨壮医滋勒调经汤改善卵巢储备功能低下的作用机制。方法:选取性成熟、动情周期正常的70只雌性SD大鼠,用随机方法分为造模组(60只)和空白对照组(10只)。造模组予雷公藤多甙(gluc-osida tripterygii,TWP)按照50mg/(kg·d)剂量,15ml(kg·d)体积灌胃2周造模,自灌胃的第6天始,每天行阴道脱落细胞学涂片观察记录,对大鼠模型进行评价。造模成功后,用随机方法将造模组分为TWP模型组、坤泰胶囊组、补佳乐组、滋勒调经汤低剂量组、滋勒调经汤中剂量组、滋勒调经汤高剂量组,连续15天灌胃给药干预,并行阴道脱落细胞学涂片观察记录。治疗结束后分批将大鼠麻醉,腹主动脉取血分离血清,立即摘除卵巢及子宫称重并甲醛固定。观察大鼠存活及一般情况、体重、动情周期、子宫及卵巢指数、卵巢病理组织形态,用放免法测各组大鼠血清(follicle-stimulating hormone,FSH)、黄体生成素(luteinizing hormone,LH)、雌二醇激素(estradiol,E_2)水平,用免疫组化法检测卵巢组织中抑瘤素(oncostatin M,OSM)、巨噬细胞集落刺激因子(macrophage colony stimulating factor,M-CSF)水平表达。结果:1.TWP模型组动情周期数远低于空白对照组,差异具有统计学意义(P0.001),各治疗组动情周期数较TWP模型组比均不同程度增加,差异具有统计学意义(P0.05),其中滋勒调经汤高剂量组动情周期数最接近空白对照组。2.与空白对照组比较,TWP模型组大鼠卵巢、子宫指数均较低,差异具有统计学意义(P0.05,P0.001)。各治疗组同TWP模型组比,卵巢、子宫指数均不同程度增加,差异均具有统计学意义(P0.05,P0.001)。滋勒调经汤低剂量组大鼠卵巢指数低于坤泰胶囊组,且具有统计学意义(P0.05),滋勒调经汤高剂量组大鼠子宫指数高于坤泰胶囊组,差异具有统计学意义(P0.05)。滋勒调经汤低剂量组比补佳乐组大鼠子宫指数低,差异具有统计学意义(P0.05)。滋勒调经汤高剂量组卵巢、子宫指数高于低剂量组,其子宫指数高于中剂量组,差异均具有统计学意义(P0.05)。3.与空白对照组比较,TWP模型组生长卵泡、成熟卵泡及黄体均少,闭锁卵泡多,差异均具有统计学意义(P0.001),滋勒调经汤低剂量组生长卵泡、成熟卵泡及黄体数目均少于空白对照组,且差异均具有统计学意义(P0.05)。坤泰胶囊组、补佳乐组、滋勒调经汤中、高剂量组同TWP模型组比较,生长卵泡、成熟卵泡及黄体数目均有所增加,闭锁卵泡不同程度减少,差异均有统计学意义(P0.05,P0.001),滋勒调经汤低剂量组生长卵泡多于TWP模型组,差异具有统计学意义(P0.05),其成熟卵泡、黄体数目及闭锁卵泡虽有改善,但差异不具有统计学意义(P0.05)。滋勒调经汤高剂量组在成熟卵泡及黄体数目方面多于其低剂量组,差异具有统计学意义(P0.05)。4.各治疗组同TWP模型组比较,镜下见卵巢结构清晰,各级卵泡和黄体数目多,颗粒细胞层数多,与空白对照组接近,闭锁卵泡及白体少见。5.TWP模型组大鼠血清FSH、LH水平高,E_2水平低,与空白对照组比较差异具有统计学意义(P0.001),滋勒调经汤低剂量组经过治疗后,大鼠FSH水平有所下降,E_2水平有所增加,但与空白对照组相比,差异具有统计学意义(P0.05)。各治疗组大鼠经过治疗后,血清FSH、LH水平均不同程度下降,E_2水平不同程度上升,与TWP模型组比较,其中坤泰胶囊组、补佳乐组、滋勒调经汤中、高剂量组差异均有统计学意义(P0.05,P0.001)。滋勒调经汤高剂量组血清E_2水平高于其低剂量组,差异具有统计学意义(P0.05)。6.滋勒调经汤低剂量组、TWP模型组卵巢OSM IOD值与空白对照组比,差异具有统计学意义(P0.001),各治疗组卵巢OSM IOD值均明显增加,与TWP模型组相比较,差异均具有统计学意义(P0.001)。滋勒调经汤低剂量组中OSM IOD值低于坤泰胶囊组和补佳乐组,差异均具有统计学意义(P0.05)。滋勒调经汤高剂量组OSM IOD值高于其低剂量组,差异具有统计学意义(P0.05)。7.滋勒调经汤低剂量组、TWP模型组卵巢M-CSF IOD值与空白对照组比,差异具有统计学意义(P0.001),各治疗组卵巢M-CSF IOD值均明显增加,与TWP模型组相比较,差异均具有统计学意义(P0.001)。滋勒调经汤低剂量组中M-CSF IOD值低于坤泰胶囊组和滋勒调经汤高剂量组(P0.05)。结论:滋勒调经汤可增加DOR大鼠卵巢、子宫脏器指数,增加各级卵泡及黄体数目,减少闭锁卵泡,可通过降低血清FSH、LH水平,升高E_2水平来调节性激素,增加卵巢组织中OSM、M-CSF表达,促进卵泡生长发育,这可能是其防治DOR的机制之一。坤泰胶囊、补佳乐及滋勒调经汤均能改善卵巢储备功能,促进卵泡生长发育及成熟。滋勒调经汤的疗效呈一定剂量依赖性,其中剂量与坤泰胶囊、补佳乐效果最相当,是三个剂量中最适宜的剂量。
[Abstract]:Objective: through the establishment of diminished ovarian reserve (DOR) rat model and the intervention of Zi Ling Jing Jing decoction, by observing the ovarian morphology, rat sex hormone and OSM, M-CSF cytokine and other related indexes, the mechanism of Zhuang Zi Ling Jing Jing soup to improve the ovarian reserve function was preliminarily discussed. 70 female SD rats with normal mature and estrous cycles were randomly divided into a model group (60) and a blank control group (10). The model group was given gluc-osida tripterygii (TWP) in accordance with the dosage of 50mg/ (kg d), 15ml (kg d) volume for 2 weeks, and from the sixth day of the gastric perfusion, the vaginal cytological smears were observed every day. After the model was successful, the model group was divided into TWP model group, Kun Tai capsule group, bu Jia Le group, the low dose group of tisai tonifying soup, the dose group of tisai tonifying soup, the high dose group of tisai tonifying soup, 15 days continuous perfusion of the stomach and the observation records of the smear smear of the vaginal exfoliative cytology. The rats were anaesthetized and the abdominal aorta was extracted from the blood serum and immediately removed the weight of the ovary and uterus and fixed with formaldehyde. The survival and general condition of the rats, the body weight, the estrous cycle, the uterus and ovary index, the pathological tissue morphology of the ovary were observed, and the serum (follicle-stimulating hormone, FSH), and the luteinizing hormone (luteinizing hormon) were measured by radioimmunoassay. E, LH), the level of estradiol (E_2), the expression of oncostatin M (OSM) in ovarian tissue and the level of macrophage colony stimulating factor (macrophage colony stimulating factor, M-CSF) were detected by immunohistochemistry. The number of estrous cycles in the treatment group was increased in different degrees compared with the TWP model group, and the difference was statistically significant (P0.05). The number of estrous cycles in the high dose group of Taige tonifying soup was closest to that of the blank control group.2. and the blank control group. The ovarian index of the TWP model group was lower, the difference was statistically significant (P0.05, P0.001). The differences were statistically significant (P0.05, P0.001). Compared with the TWP model group, the ovary and the uterus index were all increased in different degrees, and the difference was statistically significant (P0.05, P0.001). The ovary index of the low dose group of Tzi Tong Jing Decoction group was lower than that of the Kun Tai capsule group, and had statistical significance (P0.05). The uterus index of the high dose group of Taige tonifying Decoction group was higher than that of the Kun Tai capsule group, and the difference was statistically significant. P0.05. The uterus index of the low dose group of Taige tonifying soup was lower than that of the pincy group. The difference was statistically significant (P0.05). The uterus index of the high dose group of Taigan tonifying soup was higher than the low dose group, and the uterus index was higher than that in the middle dose group. The difference was statistically significant (P0.05).3. and the blank control group, the TWP model group grew follicles, The mature follicle and corpus luteum were less, the atresia follicular follicles were many, the differences were statistically significant (P0.001). The growth of follicles in the low dose group of Taigan tonifying soup, the number of mature follicles and corpus luteum were less than that in the blank control group, and the difference was statistically significant (P0.05). The high dose group was compared with the TWP model group in the Kun Tai capsule group, the Tinca group and the tigel tonifying soup. The growth of follicles, the number of mature follicles and corpus luteum increased, the atresia follicle decreased in varying degrees, and the difference was statistically significant (P0.05, P0.001). The growth of follicles in the low dose group of Taige tonifying soup was more than that of the TWP model group. The difference was statistically significant (P0.05). The number of mature follicles, the number of corpus luteum and the atresia follicle were improved. Study significance (P0.05). The number of mature follicles and corpus luteum in the high dose group of Taige tonifying soup was more than that of the low dose group. The difference was statistically significant (P0.05) compared with the TWP model group (.4.), the ovarian structure was clear, the number of follicles and corpus luteum at all levels, the number of granulocyte layers were more, and the number of granulocytes was close to the blank control group. In the rare.5.TWP model group, the serum FSH, the LH level and the E_2 level were low, and the difference between the control group and the blank control group was statistically significant (P0.001). After the treatment, the FSH level of the rats was decreased and the E_2 level increased, but the difference was statistically significant (P0.05) compared with the blank group (P0.05). After the treatment, the level of serum FSH and LH decreased in varying degrees, and the level of E_2 increased in different degrees. Compared with the TWP model group, the differences in the high dose group of the high dose group of Kun Tai capsule group, bu Jia Le group and Taigan tonifying soup were statistically significant (P0.05, P0.001). The serum E_2 level was higher than that of the low dose group, and the difference was statistically significant. Study significance (P0.05).6. Taigan soup low dose group, TWP model group ovarian OSM IOD value and blank control group, the difference was statistically significant (P0.001), the value of OSM IOD in each group was significantly increased, compared with the TWP model group, the difference was statistically significant (P0.001). The OSM IOD in the low dose group of tgtd group was lower than the Kun Tai capsule The difference was statistically significant (P0.05). The OSM IOD value of the high dose group of Taige tonifying Decoction group was higher than that of the low dose group, the difference was statistically significant (P0.05) in the low dose group of.7. Taigan tonifying soup, the value of M-CSF IOD in the TWP model group was compared with the blank control group, the difference was statistically significant (P0.001), and the IOD value of the ovarian M-CSF in each treatment group. Compared with TWP model group, the difference was statistically significant (P0.001). The M-CSF IOD value in the low dose group was lower than that of Kun Tai capsule group and Zige tonifying Decoction (P0.05). Conclusion: Zige tonifying decoction can increase the ovary of DOR rats, the index of uterus organs, increase the number of follicles and corpus luteum at all levels, and reduce the atresia follicle. By reducing the serum FSH, LH level and increasing the level of E_2 to regulate sex hormones and increase the expression of OSM and M-CSF in the ovarian tissue and promote the growth and development of follicles, this may be one of the mechanisms for the prevention and control of DOR. The dosage was the most suitable for the three doses.

【学位授予单位】：广西中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R29;R-332

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