补肾安胎冲剂对复发性流产小鼠蜕膜组织VEGF、VEGFR2、RAS、MAPK蛋白表达影响的实验研究

发布时间：2018-05-27 04:06

本文选题：补肾安胎冲剂 + 复发性流产　；参考：《安徽中医药大学》2016年硕士论文

【摘要】：目的:通过以VEGF信号通路为切入点,观察研究补肾安胎冲剂对复发性流产模型小鼠子宫及胚胎发育情况,胚胎丢失率及检测蜕膜组织VEGF、VEGFR2、RAS、MAPK蛋白表达,明确补肾健脾中药治疗RSA在妊娠胚胎的血管重铸方面的作用机理,为中医药补肾安胎冲剂治疗RSA提供一定的理论依据。方法:以雌鼠CBA/J与雄鼠DBA/2建立复发性流产小鼠模型,雌鼠CBA/J与雄鼠BALB/c建立正常妊娠小鼠模型,雌雄鼠按照2∶1合笼交配,适应性喂养1周后。造模的妊娠小鼠共计60只,分为六组:正常妊娠组,复发性流产模型妊娠组,西药治疗组,中药低剂量治疗组,中药中剂量治疗组,中药高剂量治疗组。从检测出小鼠妊娠的第1天算起,予以灌胃处理:复发性流产模型妊娠组即是模型组,予胃中灌入稀释后的蒸馏水;西药治疗组饲喂稀释的黄体酮胶囊,黄体酮胶囊的剂量为0.4mg/kg.d;中药低剂量治疗组即是补肾安胎冲剂低剂量组,饲喂稀释后的补肾安胎冲剂,冲剂的剂量为3.9g/kg.d;中药中剂量治疗组即是补肾安胎冲剂中剂量组,予以喂入稀释后的补肾安胎冲剂,冲剂的剂量为11.7g/kg.d;中药高剂量治疗组即是补肾安胎冲剂高剂量组,饲喂稀释后的补肾安胎冲剂,冲剂的剂量为35.1g/kg.d,连续给药共计15天。小鼠末次给药24h后,予以处死,取出子宫,剖视观察胚胎发育情况,计算胚胎丢失率,用Western blot法检测妊娠小鼠蜕膜组织中VEGF、VEGFR2、RAS、MAPK的蛋白表达水平。结果:1.剖视观察见正常妊娠组小鼠子宫及胚胎发育良好,颜色呈淡红色,形态如串珠状,质地大小较均匀,宫腔内未见显著瘀血之处,子宫的着床点未见明显的出血。模型组妊娠小鼠子宫及胚胎发育较差,颜色呈黑褐色,形态呈竹节样改变,质地不均匀,宫腔内可见明显的瘀血点,或在胚胎的着床点处见到部分或完全消失的胚胎组织。西药组、补肾安胎冲剂低、中、高剂量组子宫及胚胎质地较为均匀,呈淡红色,宫腔内未见明显的出血点,胚胎发育较好。2.与正常妊娠组相比较,模型对照组胚胎丢失率显著上升(P0.05)。与复发性流产模型妊娠组相比较,西药治疗组、补肾安胎冲剂中剂量、补肾安胎冲剂高剂量组显著降低,有统计学差异(P0.05)。3.与正常妊娠组比较,复发性流产模型妊娠组VEGF表达含量显著降低(P0.01)。与复发性流产模型妊娠组比较,西药治疗组、中药低、中、高剂量治疗组VEGF表达显然升高(P0.01);与西药治疗组、中药低剂量治疗组、中药中剂量治疗组比较,中药高剂量治疗组的VEGF表达含量显著升高(P0.05)。4.与正常妊娠组比较,复发性流产模型妊娠组VEGFR2表达含量显著降低(P0.01)。与复发性流产模型妊娠组比较,西药治疗组、中药低、中、高剂量治疗组VEGFR2表达显然升高(P0.01);与西药治疗组、中药低剂量组、中药中剂量治疗组比较,中药高剂量治疗组的VEGFR2表达含量显著升高(P0.05)。5.与正常妊娠组比较,复发性流产模型妊娠组RAS表达含量显著降低(P0.01)。与复发性流产模型妊娠组比较,西药治疗组、中药低、中、高剂量治疗组RAS表达显然升高(P0.01);与西药治疗组、中药低剂量组、中药中剂量治疗组比较,中药高剂量治疗组的RAS表达含量显著升高(P0.05)。6.与正常妊娠组比较,复发性流产模型妊娠组MAPK表达含量显著降低(P0.01)。与复发性流产模型妊娠组比较,西药治疗组、中药低、中、高剂量治疗组MAPK表达显然升高(P0.01);与西药治疗组、中药低剂量组、中药中剂量治疗组比较,中药高剂量治疗组的MAPK表达含量显著升高(P0.05)。结论:1、补肾安胎冲剂对复发性流产小鼠妊娠子宫和胚胎发育可能有一定的促进作用。2、补肾安胎冲剂能显著降低复发性流产小鼠模型的胚胎丢失率,达到保胎的疗效。3、VEGF、VEGFR2、RAS、MAPK的蛋白印迹检测结果,指出补肾安胎冲剂治疗复发性流产发挥有效疗效的作用机制可能是上调VEGF信号通路下游的RAS、MAPK蛋白表达水平,同时提高复发性流产小鼠蜕膜组织的VEGF、VEGFR2蛋白表达水平,改善复发性流产模型小鼠母胎界面血管生成,维持妊娠,有效降低了复发性流产小鼠的胚胎丢失率。
[Abstract]:Objective: To observe the development of uterus and embryo, the rate of embryo loss and the expression of VEGF, VEGFR2, RAS and MAPK protein in the decidua tissue, by using the VEGF signal pathway as the breakthrough point, and to clarify the mechanism of the effect of the Chinese herbal medicine for invigorating the kidney and invigorating the spleen on the vascular recasting of pregnancy induced pregnancy. To provide a certain theoretical basis for the treatment of RSA by Bushen Antai granule. Methods: a mouse model of recurrent abortion was established in the female rat CBA/J and the male rat DBA/2. The female rat CBA/J and the male rat BALB/c established the normal pregnancy mice model. The male and female mice were copulated according to the 2: 1 cage, and after 1 weeks of adaptive feeding. The model pregnant mice were divided into six groups: normal pregnancy group, The recurrent abortion model pregnancy group, the western medicine treatment group, the traditional Chinese medicine low dose treatment group, the medium dose treatment group, the high dose Chinese medicine treatment group. From the first days of the pregnancy test, the recurrent abortion model pregnancy group was the model group, the diluted distilled water was injected into the stomach, and the western medicine treatment group was fed the diluted yellow. The dose of somatic ketone capsule, the dose of progesterone capsule is 0.4mg/kg.d, the low dose treatment group of traditional Chinese medicine is the low dose group of Bushen Antai granule, and the dosage of the diluted kidney and Antai granule is 3.9g/kg.d, the dosage group in the middle dose of Chinese traditional medicine is the dose group of the tonifying kidney antire granules, and the dosage of the decoction is given after the dilution. 11.7g/kg.d, the high dose group of traditional Chinese medicine was a high dose group of Bushen Antai granule, and the dosage of the diluted kidney and anfetus was 35.1g/kg.d, the dosage of the granule was 15 days. After the last dose of 24h, the mice were executed, the uterus was taken out, the embryo development situation was observed, the embryo loss rate was calculated, the pregnancy loss was detected by Western blot method. The pregnancy was detected by Western blot method. The protein expression level of VEGF, VEGFR2, RAS, MAPK in the rat Decidua Tissue. Results: 1. the uterus and embryo of the normal pregnancy group were well developed, the color of the uterus and the embryo were well developed, the color was light red, the shape was like the beads, the size of the texture was more uniform, the blood stasis of the uterus was not obvious in the uterus. The uterus and the uterus of the uterus were not obviously bleeding. The uterus of the model group of pregnancy mice and the uterus and the uterus and the uterus of the model group of pregnancy mice were not seen. The embryo development is poor, the color is black and brown, the shape is bamboo node change, the texture is uneven, the intrauterine obvious blood stasis is visible, or the embryo tissue which is partially or completely disappeared at the implantation point of the embryo. The western medicine group, the kidney and the high dose group of the uterus and the high dose group are more uniform, pale red, and the uterine cavity is not obvious. When compared with the normal pregnancy group, the embryo loss rate of the model control group was significantly higher than the normal pregnancy group (P0.05). Compared with the recurrent abortion model pregnancy group, the western medicine treatment group, the dosage of tonifying kidney anetin granule and the high dose group of Bushen antire granules were significantly reduced, and the difference was statistically significant (P0.05).3. was compared with the normal pregnancy group (P0.05). The expression of VEGF in the pregnant group with recurrent abortion was significantly decreased (P0.01). Compared with the recurrent abortion model pregnancy group, the expression of VEGF in the treatment group of Western medicine was obviously elevated (P0.01) in the low, middle and high dose treatment group. Compared with the western medicine treatment group, the low dose treatment group of traditional Chinese medicine, the dose treatment group in Chinese medicine, the VEGF expression in the high dose treatment group of Chinese medicine. Significantly higher (P0.05).4. compared with the normal pregnancy group, the expression of VEGFR2 in the recurrent abortion model group was significantly lower (P0.01). Compared with the recurrent abortion model pregnancy group, the western medicine treatment group, the low, middle, high dose treatment group of the Chinese medicine group obviously increased VEGFR2 expression (P0.01); with the western medicine treatment group, the low dose group of Chinese medicine, Chinese medicine medium dose treatment group Compared with the normal pregnancy group, the expression of VEGFR2 in the high dose treatment group was significantly higher (P0.05) than that in the normal pregnancy group. The expression of RAS in the recurrent abortion model group was significantly lower (P0.01). Compared with the recurrent abortion model pregnancy group, the western medicine treatment group, the low, middle and high dose treatment group of the western medicine treatment group obviously increased (P0.01), and the western medicine treatment group (P0.01); and the western medicine treatment group. Compared with the medium dose treatment group, the expression of RAS in the high dose treatment group of Chinese traditional medicine was significantly increased (P0.05).6. and the normal pregnancy group, the expression of MAPK in the recurrent abortion model group was significantly lower (P0.01). Compared with the recurrent abortion model pregnancy group, the Western Medicine treatment group, the Chinese medicine low, middle, high dose treatment group MAPK The expression of MAPK was obviously elevated (P0.01); compared with the western medicine treatment group, the low dose group of traditional Chinese medicine and the dose treatment group in the Chinese medicine, the expression of the high dose treatment group was significantly increased (P0.05). Conclusion: 1, the tonifying kidney Antai granule may have a certain promotion effect on the pregnancy uterus and embryo development of the recurrent abortion mice, and the tonifying kidney Antai granule can decrease significantly. The rate of embryo loss in mice with low recurrent abortion can achieve the curative effect of.3, VEGF, VEGFR2, RAS, MAPK, and points out that the mechanism of the effective effect of Bushen Antai granule in the treatment of recurrent abortion may be the up-regulation of the RAS, the expression of MAPK protein downstream of the VEGF signaling pathway and the increase of recurrent abortion in mice. The expression of VEGF and VEGFR2 protein in the decidual tissue improves the angiogenesis in the maternal fetal interface of recurrent abortion model mice and maintains pregnancy, which effectively reduces the rate of embryo loss in recurrent abortion mice.
【学位授予单位】：安徽中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R271

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