本文選題：補(bǔ)腎安胎沖劑 + 復(fù)發(fā)性流產(chǎn)　； 參考：《安徽中醫(yī)藥大學(xué)》2016年碩士論文

【摘要】：目的:通過(guò)以VEGF信號(hào)通路為切入點(diǎn),觀(guān)察研究補(bǔ)腎安胎沖劑對(duì)復(fù)發(fā)性流產(chǎn)模型小鼠子宮及胚胎發(fā)育情況,胚胎丟失率及檢測(cè)蛻膜組織VEGF、VEGFR2、RAS、MAPK蛋白表達(dá),明確補(bǔ)腎健脾中藥治療RSA在妊娠胚胎的血管重鑄方面的作用機(jī)理,為中醫(yī)藥補(bǔ)腎安胎沖劑治療RSA提供一定的理論依據(jù)。方法:以雌鼠CBA/J與雄鼠DBA/2建立復(fù)發(fā)性流產(chǎn)小鼠模型,雌鼠CBA/J與雄鼠BALB/c建立正常妊娠小鼠模型,雌雄鼠按照2∶1合籠交配,適應(yīng)性喂養(yǎng)1周后。造模的妊娠小鼠共計(jì)60只,分為六組:正常妊娠組,復(fù)發(fā)性流產(chǎn)模型妊娠組,西藥治療組,中藥低劑量治療組,中藥中劑量治療組,中藥高劑量治療組。從檢測(cè)出小鼠妊娠的第1天算起,予以灌胃處理:復(fù)發(fā)性流產(chǎn)模型妊娠組即是模型組,予胃中灌入稀釋后的蒸餾水;西藥治療組飼喂稀釋的黃體酮膠囊,黃體酮膠囊的劑量為0.4mg/kg.d;中藥低劑量治療組即是補(bǔ)腎安胎沖劑低劑量組,飼喂稀釋后的補(bǔ)腎安胎沖劑,沖劑的劑量為3.9g/kg.d;中藥中劑量治療組即是補(bǔ)腎安胎沖劑中劑量組,予以喂入稀釋后的補(bǔ)腎安胎沖劑,沖劑的劑量為11.7g/kg.d;中藥高劑量治療組即是補(bǔ)腎安胎沖劑高劑量組,飼喂稀釋后的補(bǔ)腎安胎沖劑,沖劑的劑量為35.1g/kg.d,連續(xù)給藥共計(jì)15天。小鼠末次給藥24h后,予以處死,取出子宮,剖視觀(guān)察胚胎發(fā)育情況,計(jì)算胚胎丟失率,用Western blot法檢測(cè)妊娠小鼠蛻膜組織中VEGF、VEGFR2、RAS、MAPK的蛋白表達(dá)水平。結(jié)果:1.剖視觀(guān)察見(jiàn)正常妊娠組小鼠子宮及胚胎發(fā)育良好,顏色呈淡紅色,形態(tài)如串珠狀,質(zhì)地大小較均勻,宮腔內(nèi)未見(jiàn)顯著瘀血之處,子宮的著床點(diǎn)未見(jiàn)明顯的出血。模型組妊娠小鼠子宮及胚胎發(fā)育較差,顏色呈黑褐色,形態(tài)呈竹節(jié)樣改變,質(zhì)地不均勻,宮腔內(nèi)可見(jiàn)明顯的瘀血點(diǎn),或在胚胎的著床點(diǎn)處見(jiàn)到部分或完全消失的胚胎組織。西藥組、補(bǔ)腎安胎沖劑低、中、高劑量組子宮及胚胎質(zhì)地較為均勻,呈淡紅色,宮腔內(nèi)未見(jiàn)明顯的出血點(diǎn),胚胎發(fā)育較好。2.與正常妊娠組相比較,模型對(duì)照組胚胎丟失率顯著上升(P0.05)。與復(fù)發(fā)性流產(chǎn)模型妊娠組相比較,西藥治療組、補(bǔ)腎安胎沖劑中劑量、補(bǔ)腎安胎沖劑高劑量組顯著降低,有統(tǒng)計(jì)學(xué)差異(P0.05)。3.與正常妊娠組比較,復(fù)發(fā)性流產(chǎn)模型妊娠組VEGF表達(dá)含量顯著降低(P0.01)。與復(fù)發(fā)性流產(chǎn)模型妊娠組比較,西藥治療組、中藥低、中、高劑量治療組VEGF表達(dá)顯然升高(P0.01);與西藥治療組、中藥低劑量治療組、中藥中劑量治療組比較,中藥高劑量治療組的VEGF表達(dá)含量顯著升高(P0.05)。4.與正常妊娠組比較,復(fù)發(fā)性流產(chǎn)模型妊娠組VEGFR2表達(dá)含量顯著降低(P0.01)。與復(fù)發(fā)性流產(chǎn)模型妊娠組比較,西藥治療組、中藥低、中、高劑量治療組VEGFR2表達(dá)顯然升高(P0.01);與西藥治療組、中藥低劑量組、中藥中劑量治療組比較,中藥高劑量治療組的VEGFR2表達(dá)含量顯著升高(P0.05)。5.與正常妊娠組比較,復(fù)發(fā)性流產(chǎn)模型妊娠組RAS表達(dá)含量顯著降低(P0.01)。與復(fù)發(fā)性流產(chǎn)模型妊娠組比較,西藥治療組、中藥低、中、高劑量治療組RAS表達(dá)顯然升高(P0.01);與西藥治療組、中藥低劑量組、中藥中劑量治療組比較,中藥高劑量治療組的RAS表達(dá)含量顯著升高(P0.05)。6.與正常妊娠組比較,復(fù)發(fā)性流產(chǎn)模型妊娠組MAPK表達(dá)含量顯著降低(P0.01)。與復(fù)發(fā)性流產(chǎn)模型妊娠組比較,西藥治療組、中藥低、中、高劑量治療組MAPK表達(dá)顯然升高(P0.01);與西藥治療組、中藥低劑量組、中藥中劑量治療組比較,中藥高劑量治療組的MAPK表達(dá)含量顯著升高(P0.05)。結(jié)論:1、補(bǔ)腎安胎沖劑對(duì)復(fù)發(fā)性流產(chǎn)小鼠妊娠子宮和胚胎發(fā)育可能有一定的促進(jìn)作用。2、補(bǔ)腎安胎沖劑能顯著降低復(fù)發(fā)性流產(chǎn)小鼠模型的胚胎丟失率,達(dá)到保胎的療效。3、VEGF、VEGFR2、RAS、MAPK的蛋白印跡檢測(cè)結(jié)果,指出補(bǔ)腎安胎沖劑治療復(fù)發(fā)性流產(chǎn)發(fā)揮有效療效的作用機(jī)制可能是上調(diào)VEGF信號(hào)通路下游的RAS、MAPK蛋白表達(dá)水平,同時(shí)提高復(fù)發(fā)性流產(chǎn)小鼠蛻膜組織的VEGF、VEGFR2蛋白表達(dá)水平,改善復(fù)發(fā)性流產(chǎn)模型小鼠母胎界面血管生成,維持妊娠,有效降低了復(fù)發(fā)性流產(chǎn)小鼠的胚胎丟失率。
[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.
【學(xué)位授予單位】：安徽中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類(lèi)號(hào)】：R271

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