發(fā)布時間：2018-06-09 20:37

  本文選題：壽胎丸 + 滋養(yǎng)層細(xì)胞�。� 參考：《南方醫(yī)科大學(xué)》2017年碩士論文

【摘要】：研究背景及目的自然流產(chǎn)(Spontaneous abortion,SA)是妊娠常見并發(fā)癥,按自然流產(chǎn)發(fā)展的不同階段,可分為先兆流產(chǎn)、難免流產(chǎn)、不全流產(chǎn)、完全流產(chǎn)。對自然流產(chǎn)患者及時有效的治療,可進(jìn)一步影響妊娠結(jié)局。故自然流產(chǎn)預(yù)防及治療所展開的研究具有一定的社會意義。清代醫(yī)家張錫純在《醫(yī)學(xué)衷中參西錄》創(chuàng)立的壽胎丸方現(xiàn)廣泛用于自然流產(chǎn)的防治。前期實(shí)驗(yàn)研究發(fā)現(xiàn)壽胎丸含藥血清呈劑量依賴性地增強(qiáng)自然流產(chǎn)患者滋養(yǎng)層細(xì)胞的增殖活性、侵襲遷移力,減少細(xì)胞凋亡。基于此,本課題探討壽胎丸含藥血清對自然流產(chǎn)患者滋養(yǎng)層細(xì)胞凋亡及凋亡蛋白Bcl-2、Bax、Fas、FasL、Survivin表達(dá)的影響,以期進(jìn)一步探討壽胎丸防治自然流產(chǎn)作用機(jī)制及其作用靶點(diǎn)。研究方法1、將36只SPF級雌性SD大鼠隨機(jī)分為3組,分別予壽胎丸水煎液(6.25g/kg)、地屈孕酮水溶液(2.08 mg/kg)、等量生理鹽水灌胃,制備壽胎丸含藥血清、地屈孕酮含藥血清及空白血清。2、體外分離培養(yǎng)正常妊娠人工流產(chǎn)者及難免流產(chǎn)患者滋養(yǎng)層細(xì)胞(Trophoblastic cells,TB),采用免疫細(xì)胞化學(xué)法檢測貼壁細(xì)胞中細(xì)胞角蛋白(Cytokeratin-7,CK-7)和波形蛋白(Vimentin)表達(dá),證實(shí)為滋養(yǎng)層細(xì)胞后進(jìn)行后續(xù)實(shí)驗(yàn)。3、不同濃度(5%、10%、20%)壽胎丸含藥血清干預(yù)自然流產(chǎn)患者滋養(yǎng)層細(xì)胞,空白血清及地屈孕酮含藥血清分別作為空白對照及陽性對照。4、采用流式細(xì)胞術(shù)(Flow cytometry,FCM)檢測各組滋養(yǎng)層細(xì)胞凋亡情況。5、采用蛋白質(zhì)免疫印跡法(Western blot)檢測各組凋亡相關(guān)蛋白Bcl-2、Bax、Fas、FasL、Survivin表達(dá)的差異并分析其相關(guān)性。6、統(tǒng)計(jì)學(xué)方法:采用SPSS20.0統(tǒng)計(jì)學(xué)軟件進(jìn)行分析,計(jì)量資料以(x±s)表示。根據(jù)實(shí)驗(yàn)設(shè)計(jì)的不同采用不同統(tǒng)計(jì)方法,以P0.05為差異有統(tǒng)計(jì)學(xué)意義。研究結(jié)果1、與正常對照組比較,自然流產(chǎn)患者滋養(yǎng)層細(xì)胞早期凋亡、總凋亡比例明顯升高(P0.05)。2、與正常對照組相比,自然流產(chǎn)患者滋養(yǎng)層細(xì)胞Bcl-2、Survivin蛋白表達(dá)量減少,Fas、FasL、Bax蛋白表達(dá)量增多(P0.01)。3、與空白血清對照組相比,壽胎丸含藥血清、地屈孕酮含藥血清能降低自然流產(chǎn)患者滋養(yǎng)層細(xì)胞早期凋亡、總凋亡比例(p0.01)。4、與空白血清對照組相比,壽胎丸含藥血清、地屈孕酮含藥血清能上調(diào)Bcl-2、Survivin蛋白水平,下調(diào)Fas、FasL、Bax蛋白水平(P0.01)。5、壽胎丸含藥血清對自然流產(chǎn)患者滋養(yǎng)層細(xì)胞凋亡及凋亡蛋白的影響呈劑量依賴性(p0.05)。6、Bcl-2與Survivin存在密切的正相關(guān)關(guān)系,Bax、Fas、FasL與Survivin存在密切的負(fù)相關(guān)關(guān)系(P0.01)。結(jié)論壽胎丸含藥血清呈劑量依賴性減少自然流產(chǎn)患者滋養(yǎng)層細(xì)胞凋亡,上調(diào)Bcl-2、Survivin蛋白,下調(diào)Fas、FasL、Bax蛋白,從內(nèi)源性途徑、外源性途徑抑制滋養(yǎng)層細(xì)胞凋亡是壽胎丸防治自然流產(chǎn)可能作用機(jī)制之一。
[Abstract]:Background and objective spontaneous abortion is a common complication of pregnancy. It can be divided into threatened abortion, incomplete abortion and complete abortion according to different stages of the development of spontaneous abortion. Timely and effective treatment of spontaneous abortion patients can further affect the outcome of pregnancy. Therefore, the study of prevention and treatment of spontaneous abortion has certain social significance. Zhang Xichun, a doctor in Qing Dynasty, founded Shou-Tye Pill prescription in Shen Xilu, widely used in the prevention and treatment of spontaneous abortion. In previous studies, it was found that the serum of Shoufeiwan enhanced the proliferation activity, invasion and migration of trophoblastic cells in a dose-dependent manner, and reduced the apoptosis of trophoblast cells in a dose-dependent manner. Based on this, the purpose of this study was to investigate the effect of Shou-Feiwan on the apoptosis of trophoblastic cells and the expression of apoptosis protein Bcl-2OBax-Fas-FasL+ survivin in the trophoblast of spontaneous abortion patients, in order to further explore the mechanism and targets of Shou-Feiwan in the prevention and treatment of spontaneous abortion. Methods 1. Thirty-six SPF female Sprague-Dawley rats were randomly divided into 3 groups. They were given Shou-Tye Pill decoction (6.25g / kg / kg), Diproprogesterone solution (2.08 mg / kg), and the same amount of normal saline intragastric perfusion to prepare the serum of Shou-Tye Pill. Trophoblastic cells were isolated from trophoblastic cells of normal pregnancy induced abortion and inevitable abortion in vitro. Cytokeratin cytokeratin-7 (CK-7) and vimentin (Vimentin) in adherent cells were detected by immunocytochemistry. It was confirmed that for the trophoblastic cells, the following experiments were carried out. 3. Different concentrations of the trophoblastic cells were used to intervene in the trophoblast cells of the spontaneous abortion patients with the serum of Shou-Feiwan. Blank serum and prodroxyprogesterone serum were used as blank control and positive control respectively. Flow cytometry (FCM) was used to detect the apoptosis of trophoblastic cells in each group. Western blot was used to detect apoptosis correlation in each group. The difference of expression of FasLN survivin in FasL of Bcl-2P and FasL was analyzed and its correlation was analyzed. 6. Statistical method: SPSS 20.0 statistical software was used to analyze the expression of survivin, and the expression of survivin was analyzed by SPSS 20.0 software. The metrological data are expressed as x 鹵s. According to the different design of the experiment using different statistical methods, with P0.05 as the difference has statistical significance. Results 1.Compared with the normal control group, the early apoptosis of trophoblastic cells in patients with spontaneous abortion was significantly higher than that in the normal control group, and the ratio of total apoptosis was significantly higher than that in the normal control group. The expression of Bcl-2P survivin protein in trophoblast cells of patients with spontaneous abortion was decreased. The expression of Fas-FasL- Bax protein was increased. Compared with the control group, the serum of Shou-Feiwan and Divergesterone could reduce the early apoptosis of trophoblast cells in the patients with spontaneous abortion, compared with the control group, Shoufeiwan and Diproprogesterone could reduce the early apoptosis of trophoblastic cells. Compared with the control group, the serum of Shou-Li Wan and Diproprogone could upregulate the level of Bcl-2P survivin protein. The level of FasL / Bax protein was down-regulated (P 0.01). 5. The effect of Shouti pills on the apoptosis and apoptotic protein of trophoblast cells in spontaneous abortion patients was found to be in a dose-dependent manner. There was a close positive correlation between FasL and survivin. There was a close negative correlation between FasL and survivin (P 0.01). Conclusion the serum of Shou-Feiwan can decrease the apoptosis of trophoblastic cells in a dose-dependent manner, up-regulate the expression of Bcl-2 survivin protein and down-regulate Fas-FasL- Bax protein in spontaneous abortion patients. Inhibition of trophoblastic cell apoptosis by exogenous pathway is one of the possible mechanisms of Shou-Feiwan in the prevention and treatment of spontaneous abortion.
【學(xué)位授予單位】：南方醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R714.21

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 李亞;劉新玉;王俊玲;劉昱磊;滕輝;;壽胎丸含藥血清對自然流產(chǎn)患者滋養(yǎng)層細(xì)胞生物學(xué)特性的影響[J];中國中西醫(yī)結(jié)合雜志;2016年05期

2 陳健媚;郭姣;;中藥血清藥理學(xué)研究進(jìn)展[J];廣東藥學(xué)院學(xué)報(bào);2016年03期

3 史銀花;;壽胎丸治療習(xí)慣性流產(chǎn)臨床研究[J];亞太傳統(tǒng)醫(yī)藥;2016年05期

4 馬云秀;楊國嶸;郭群;張巍巍;趙甲升;;早期自然流產(chǎn)患者胎盤絨毛滋養(yǎng)細(xì)胞中Fas、Survivin、Ki-67的表達(dá)及意義[J];中國優(yōu)生與遺傳雜志;2015年07期

5 白雪;李冬梅;郝妍;鄭春楊;尹江峰;;實(shí)時熒光定量法檢測壽胎丸對實(shí)驗(yàn)性流產(chǎn)大鼠的安胎作用[J];中國婦幼保健;2015年13期

6 馬亦良;黃娟;白軍;;胎盤細(xì)胞凋亡與早產(chǎn)流產(chǎn)關(guān)系研究[J];延安大學(xué)學(xué)報(bào)(醫(yī)學(xué)科學(xué)版);2014年01期

7 李慧芳;雷磊;譚展望;羅蕾;盧麗麗;;壽胎丸對復(fù)發(fā)性流產(chǎn)模型小鼠子宮蛻膜HSP27和Tf表達(dá)的影響[J];中華中醫(yī)藥雜志;2014年03期

8 李肖鳳;李偉莉;張濤;;300例先兆流產(chǎn)中醫(yī)證候調(diào)查分析[J];安徽中醫(yī)藥大學(xué)學(xué)報(bào);2014年01期

9 司馬玲;姜群英;馮桂萍;蔡霞;;反復(fù)自然流產(chǎn)絨毛滋養(yǎng)層細(xì)胞中survivin、Bax的表達(dá)及其與細(xì)胞凋亡的關(guān)系研究[J];細(xì)胞與分子免疫學(xué)雜志;2013年11期

10 孫宇;趙紅;;自然流產(chǎn)患者心理問題及其干預(yù)的研究現(xiàn)狀[J];中華護(hù)理雜志;2013年07期

相關(guān)博士學(xué)位論文 前3條

1 繆江霞;續(xù)斷有效部位的提取及其補(bǔ)腎安胎的藥效學(xué)研究[D];廣州中醫(yī)藥大學(xué);2012年

2 黃長盛;桑寄生及其有效部位的補(bǔ)腎安胎藥效學(xué)研究[D];廣州中醫(yī)藥大學(xué);2012年

3 劉新玉;菟絲子及其有效部位提取物補(bǔ)腎安胎藥效研究[D];廣州中醫(yī)藥大學(xué);2012年

相關(guān)碩士學(xué)位論文 前2條

1 宋錕;早期先兆流產(chǎn)的中醫(yī)證型分布及其與孕酮、β-hCG的相關(guān)性研究[D];北京中醫(yī)藥大學(xué);2016年

2 張列轉(zhuǎn);自然流產(chǎn)小鼠胎盤細(xì)胞凋亡及其相關(guān)調(diào)控基因Fas/FasL，，Bcl-2/Bax的表達(dá)[D];山西醫(yī)科大學(xué);2007年

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