肾虚痰湿型多囊卵巢综合征病证结合动物模型的建立研究

发布时间：2018-02-28 19:12

本文关键词： 来曲唑脱氢表雄酮大鼠模型肾虚痰湿肥胖多囊卵巢综合征　出处：《成都中医药大学》2016年硕士论文　论文类型：学位论文

【摘要】：[实验目的]筛选出符合具有中医肾虚痰湿特征信息PCOS病证结合即临床与代谢特征的最优动物模型。并研究Bax、Bcl-2、GDF-9、BMPR-2在PCOS大鼠卵巢中的表达水平,初步探讨肥胖型PCOS发病机制。[实验方法]健康42d龄、23d龄SD雌性大鼠各70只,连续5天行阴道分泌物涂片检查,选取动情周期正常的为入组标准。将42d龄大鼠随机分为空白对照组、高脂模型组、Letrozole模型组、Letrozole肥胖模型组；将23d龄大鼠随机分为空白对照组、高脂模型组、DHEA模型组、DHEA肥胖模型组。分别在Letrozole、 DHEA基础上加用高脂乳剂,连续造模处理21天后,通过观察大鼠一般情况、动情周期,检测大鼠用药前后体重差、脏器指数、胰岛素(FINS、2hINS、FPG、 2hBPG、HOMA指数)水平、血清性激素(E2、T、LH、FSH、LH/FSH)水平、血脂(TC、TG、HDL、LDL)水平并将卵巢送检。HE染色后,在显微镜下观察各组大鼠卵巢组织,用免疫组化法检测Bax、Bcl-2、GDF-9、BMPR-2在PCOS大鼠卵巢中的表达水平。[实验结果](1)对动情周期的影响Letrozole模型组、Letrozole肥胖模型组、DHEA模型组、DHEA肥胖模型组大鼠均出现动情周期紊乱,或动情周期延长,或持续处于动情期。大鼠排卵率明显降低,差异有统计学意义(P0.01)。(2)对体重的影响Letrozole模型组、Letrozole肥胖模型组、DHEA模型组、DHEA肥胖模型组大鼠用药前后体重差及大鼠处死时体重升高明显,差异有统计学意义(P0.01)。(3)对脏器指数的影响Letrozole模型组、Letrozole肥胖模型组大鼠子宫脏器指数下降明显,卵巢脏器指数升高明显,差异有统计学意义(P0.01)。DHEA模型组、DHEA肥胖模型组大鼠子宫脏器指数升高,差异有统计学意义(P0.05)。(4)对血糖和胰岛素的影响Letrozole模型组、Letrozole肥胖模型组大鼠FINS、FPG值升高,差异有统计学意义(P0.05),2hFINS升高明显,差异有统计学意义(P0.01)。DHEA模型组、DHEA肥胖模型组大鼠FINS. HOMA指数升高,差异有统计学意义(P 0.05),2hFINS升高明显,差异有统计学意义(P0.01)。(5)对性激素的影响Letrozole;模型组、Letrozole肥胖模型组大鼠E2、LH/FSH值升高,差异有统计学意义(P0.05),T、LH值升高明显,差异有统计学意义(P0.01)。DHEA模型组、DHEA肥胖模型组大鼠E2、T、LH、FSH值升高明显,差异有统计学意义(P0.01)。(6)对血脂的影响Letrozole模型组、Letrozole肥胖模型组大鼠TC、TG值升高,HDL值降低,差异有统计学意义(P0.05)。DHEA模型组、DHEA肥胖模型组大鼠TC、 TG值升高,差异有统计学意义(P0.05)。(7)对卵巢形态学影响Letrozole模型组、Letrozole肥胖模型组大鼠卵巢初级卵泡数、次级卵泡数、成熟卵泡数、黄体数均减少,差异有统计学意义(P0.05),囊状卵泡数增多,差异有统计学意义(P0.05),闭锁卵泡数增多明显,差异有统计学意义(P＜0.01)。DHEA模型组、DHEA肥胖模型组大鼠卵巢成熟卵泡数、黄体数减少,差异有统计学意义(P0.05),闭锁卵泡数增多明显,差异有统计学意义(P0.01)。(8) Bax、Bcl-2的表达研究Letrozole模型组、Letrozole肥胖模型组、DHEA模型组、DHEA肥胖模型组均出现Bax呈现高水平表达,Bc1-2呈现低水平表达,差异无统计学意义(P0.05)。(9) GDF-9、BMPR-2的表达研究Letrozole模型组、Letrozole肥胖模型组、DHEA模型组、DHEA肥胖模型组GDF-9、BMPR-2均呈现低水平表达,差异无统计学意义(P0.05)[实验结论](1) Letrozole改良法和DHEA改良法均较符合具有中医肾虚痰湿特征信息病证结合PCOS动物模型。(2) Letrozole改良法偏重于研究PCOS大鼠血脂、胰岛素抵抗、卵巢形态学改变。(3)DHEA改良法偏重于研究PCOS大鼠血清性激素水平、卵巢形态学改变。(4)推测Bax在肥胖型PCOS动物模型中呈现高水平表达,Bcl-2、GDF-9、 BMPR-2均呈现低水平表达。
[Abstract]:Objective : To screen out the optimal animal model for syndrome of PCOS ( PCOS ) with syndrome of kidney - deficiency and phlegm - dampness syndrome ( PCOS ) , and to study the expression level of Bax , Bcl - 2 , GDF - 9 and BMPR - 2 in the ovary of PCOS rats . The rats were randomly divided into blank control group , high fat model group , Letrozole model group and Letrozole obesity model group . The rats were randomly divided into blank control group , high fat model group , Letrozole model group and Letrozole obesity model group . The rats were randomly divided into blank control group , high fat model group , Letrozole model group and Letrozole obesity model group . After HE staining , the expression level of Bax , Bcl - 2 , GDF - 9 and BMPR - 2 in the ovary of PCOS rats was observed by immunohistochemistry . ( 2 ) The effects of the Letrozole model group , the Letrozole obesity model group , the model group of Letrozole , the model group of Letrozole , the model group of Letrozole , the model group of Letrozole , the model group of Letrozole in obese model group , the model group of Letrozole obesity model group , the model group of Letrozole obesity model group , the model group of Letrozole obesity model group and the model group of Letrozole obese rats were significantly higher than those in the model group of Letrozole model group , the difference was significant ( P0.01 ) . There was significant difference in serum levels of E2 , LH and FSH in the model group and Letrozole obesity model group ( P0.05 ) . The expression of Bax and BMPR - 2 in obese model group was significantly higher than that in Letrozole model group ( P0.05 ) . ( 4 ) Bcl - 2 , GDF - 9 and BMPR - 2 were expressed at low level in obese women with PCOS .

【学位授予单位】：成都中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R271.9

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