本文選題：雷公藤甲素 + Ntoch信號(hào)通路��； 參考：《青島大學(xué)》2017年碩士論文

【摘要】：目的:通過實(shí)驗(yàn)觀察雷公藤甲素對(duì)阿霉素腎病(ADR)大鼠足細(xì)胞凋亡的影響并初步探討雷公藤甲素減輕足細(xì)胞損傷的作用機(jī)制。方法:將45只雄性Wistar大鼠隨機(jī)分為對(duì)照組(N)、阿霉素腎病模型組(ADR)及雷公藤干預(yù)組(T)。其中ADR組和T組大鼠經(jīng)尾靜脈一次性注射阿霉素6.5mg/kg構(gòu)建阿霉素腎病大鼠模型,而N組大鼠則經(jīng)尾靜脈一次性注射同體積的生理鹽水。模型建立后,T組給予雷公藤甲素(200ug/kg·d)干預(yù)8周。于2、4、6、8周分別檢測(cè)24h尿蛋白定量(24h UTP),于8周末采血,觀察各組大鼠的生化指標(biāo)如血尿素氮(BUN)、肌酐(Scr)、胱抑素C(Cys C),并在光鏡、電鏡下觀察腎組織學(xué)改變,應(yīng)用TUNEL法檢測(cè)足細(xì)胞凋亡,免疫組織化學(xué)染色檢測(cè)腎母細(xì)胞瘤基因(WT-1)半定量計(jì)數(shù)足細(xì)胞,實(shí)時(shí)熒光定量聚合酶鏈反應(yīng)(RT-PCR)檢測(cè)足細(xì)胞標(biāo)志物突觸極蛋白Synaptopodin、凋亡相關(guān)因子p53及Notch1 m RNA的表達(dá)變化,蛋白質(zhì)印記法(western)測(cè)定Synaptopodin、Notch1、Hes1的蛋白表達(dá)。結(jié)果:(1)ADR組各時(shí)間點(diǎn)24h尿蛋白定量、BUN、Scr以及Cys C指標(biāo)均高于N組,差異有統(tǒng)計(jì)學(xué)意義(P0.05);而足細(xì)胞密度顯著降低(P0.05),腎小球系膜區(qū)增寬,且系膜細(xì)胞增生;T組Scr、Cys C、24h尿蛋白定量低于ADR組(P0.05);足細(xì)胞密度明顯增加(P0.05);同時(shí)觀察到腎小球病理改變程度較ADR組減輕。(2)8周末時(shí),ADR組Synaptopodin m RNA和蛋白表達(dá)降低,p53 m RNA表達(dá)升高,足細(xì)胞體積腫脹增大,凋亡足細(xì)胞數(shù)目比N組明顯增多(P0.05);與ADR組相比,T組Synaptopodin m RNA和蛋白表達(dá)增加,p53 m RNA表達(dá)下降,凋亡足細(xì)胞數(shù)目較ADR組明顯減少(P0.05)。(3)結(jié)合RT-PCR及western結(jié)果顯示ADR組Notch1 m RNA和蛋白的表達(dá)以及Hes1蛋白的表達(dá)均高于N組(P0.05),T組Notch1 m RNA和蛋白的表達(dá)及Hes1蛋白的表達(dá)低于ADR組(P0.05)。結(jié)論:1.阿霉素腎病大鼠模型中存在足細(xì)胞損傷、凋亡;在大鼠足細(xì)胞凋亡數(shù)增加的同時(shí)Notch信號(hào)通路的相關(guān)指標(biāo)Notch1、靶基因Hes1的表達(dá)量增加,而且凋亡相關(guān)因子p53的表達(dá)同樣增加,而足細(xì)胞標(biāo)志蛋白Synaptopodin表達(dá)減少;2.Notch信號(hào)通路可能通過激活其靶基因Hes,調(diào)節(jié)凋亡相關(guān)因子p53誘導(dǎo)足細(xì)胞的凋亡;3.應(yīng)用雷公藤甲素干預(yù)后發(fā)現(xiàn),足細(xì)胞凋亡數(shù)較阿霉素腎病模型組減少,而Notch信號(hào)通路的表達(dá)也相對(duì)下降;造成阿霉素腎病足細(xì)胞凋亡的原因可能與Notch信號(hào)通路過表達(dá)有關(guān),而雷公藤甲素可能通過抑制Notch信號(hào)通路過表達(dá)從而減輕足細(xì)胞凋亡。
[Abstract]:Aim: to observe the effect of triptolide on apoptosis of podocyte in adriamycin nephropathy rats and to explore the mechanism of triptolide on podocyte injury. Methods: 45 male Wistar rats were randomly divided into control group, adriamycin nephropathy model group and tripterygium wilfordii intervention group. ADR group and T group rats were injected adriamycin 6.5mg/kg into caudal vein to establish the rat model of adriamycin nephropathy, while group N rats were injected with the same volume of normal saline via tail vein. After the model was established, T group was treated with triptolide (200ugkg / d) for 8 weeks. Blood samples were collected at the end of 8 weeks to observe the biochemical parameters such as blood urea nitrogen bun, creatinine cinnamon, cystatin C(Cys cleave, renal histologic changes under light microscope and electron microscope, and the apoptosis of podocyte by TUNEL method. Immunohistochemical staining was used to detect the semi-quantitative counting of podocytes, and real-time fluorescence quantitative polymerase chain reaction (RT-PCR- PCR) was used to detect the expression of Synaptopodin, apoptosis-related factor p53 and Notch1 m RNA. The protein expression of Synaptopodin Notch1 Hes1 was determined by Western blot. Results the levels of BUNC and Cys C in 24 h urinary protein quantitation in the 1 / 1 ADR group were significantly higher than those in N group (P 0.05), while the density of podocyte decreased significantly and the glomerular Mesangial area widened. In addition, the 24-hour urinary protein quantity of Scr-Cys Cnn in Mesangial cell proliferation T group was lower than that of ADR group (P 0.05), and the density of podocyte was significantly increased compared with that of ADR group. At the end of 8 weeks, the expression of Synaptopodin m RNA and protein in ADR group was decreased, and the expression of p53 m RNA was decreased. The number of apoptotic podocytes in podocyte was significantly increased than that in N group (P 0.05), the expression of Synaptopodin m RNA and protein in T group was higher than that in ADR group, and the expression of p53 m RNA was decreased in T group. The expression of Notch1 m RNA and protein and the expression of Hes1 protein in ADR group were significantly higher than those in group N (P 0.05). The expression of Notch1 m RNA and protein and the expression of Hes1 protein were significantly lower in ADR group than in ADR group (P 0.05). The number of apoptotic podocytes was significantly lower than that in ADR group. The results showed that the expression of Notch1 m RNA and protein and the expression of Hes1 protein in ADR group were significantly higher than those in N group. Conclusion 1. In the rat model of adriamycin nephropathy, podocytes were injured and apoptotic, and the expression of target gene Hes1 and apoptosis-related factor p53 was also increased when the number of apoptosis in podocyte increased and the related index of Notch signaling pathway Notch1 was increased, and the expression of apoptosis-related factor p53 was also increased in the rat model of adriamycin nephropathy. The decrease of Synaptopodin expression of podocyte marker protein 2. Notch signaling pathway may regulate apoptosis induced by apoptosis-related factor p53 by activating its target gene Hes3. After the intervention of triptolide, it was found that the number of podocyte apoptosis was lower than that of adriamycin nephropathy model group, and the expression of Notch signal pathway was also relatively decreased. The possible cause of podocyte apoptosis in doxorubicin nephropathy was the overexpression of Notch signal pathway. Triptolide may reduce podocyte apoptosis by inhibiting overexpression of Notch signaling pathway.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R692

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 張全樂;張紅霞;黃海長(zhǎng);章友康;;特發(fā)性足細(xì)胞病發(fā)病機(jī)制研究進(jìn)展[J];中國(guó)中西醫(yī)結(jié)合腎病雜志;2016年07期

2 黃逸輝;于力;王輝陽;于生友;蔣羅;張瑤;;足細(xì)胞相關(guān)分子在多柔比星腎病大鼠模型中mRNA的表達(dá)及意義[J];中華實(shí)用兒科臨床雜志;2016年05期

3 劉麗秋;王珂;韓潤(rùn)鴻;宋起;;雷公藤甲素對(duì)PAN致足細(xì)胞損傷中自噬作用的影響[J];中國(guó)中西醫(yī)結(jié)合腎病雜志;2015年12期

4 萬磊;劉健;黃傳兵;諶曦;汪元;張皖東;劉磊;程園園;馮云霞;;雷公藤甲素對(duì)佐劑關(guān)節(jié)炎肺功能降低大鼠Notch通路受體和配體基因表達(dá)的影響[J];南方醫(yī)科大學(xué)學(xué)報(bào);2015年10期

5 朱影;薛駿;;他克莫司治療足細(xì)胞病的作用機(jī)制的研究進(jìn)展[J];國(guó)際泌尿系統(tǒng)雜志;2015年05期

6 朱彩鳳;朱斌;包自陽;李先法;陳洪宇;沈豐平;;雷公藤內(nèi)酯醇對(duì)IgA腎病大鼠蛋白尿和nephrin、podocin蛋白及mRNA表達(dá)的影響[J];中國(guó)中西醫(yī)結(jié)合腎病雜志;2015年02期

7 張金榮;王明軍;;足細(xì)胞損傷與蛋白尿發(fā)病機(jī)理研究進(jìn)展[J];臨床腎臟病雜志;2014年06期

8 袁琴;王秋月;;Notch信號(hào)轉(zhuǎn)導(dǎo)通路與糖尿病腎病的關(guān)系[J];國(guó)際內(nèi)分泌代謝雜志;2014年02期

9 支勇;曹式麗;;雷公藤治療腎臟病作用機(jī)制研究進(jìn)展[J];山西中醫(yī);2014年03期

10 史偉;談曉凡;趙星辰;陳源漢;;足細(xì)胞損傷信號(hào)通路的研究進(jìn)展[J];中華腎病研究電子雜志;2013年06期

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