本文選題：阿魏酸 + 對(duì)乙酰氨基酚　； 參考：《重慶醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:研究阿魏酸(Ferulic acid,FA)對(duì)對(duì)乙酰氨基酚(acetaminophen,APAP)誘導(dǎo)的肝毒性的保護(hù)作用并探討其機(jī)制。方法:將Balb/c小鼠隨機(jī)分成六組,分別為空白對(duì)照組、阿魏酸對(duì)照組、模型組,低、中、高劑量的阿魏酸干預(yù)組。禁食過夜的小鼠腹腔注射APAP(350 mg/kg)建立急性肝損傷模型;分別給予10、30、100 mg/kg的阿魏酸每隔8h一次,連續(xù)灌胃3次,然后腹腔注射APAP,作為低、中、高劑量的阿魏酸干預(yù)組;以100 mg/kg的阿魏酸每隔8h一次,連續(xù)灌胃3次作為阿魏酸對(duì)照組。給予APAP18 h后檢測(cè)血清丙氨酸轉(zhuǎn)氨酶(alanine aminotransferase,ALT)、天冬氨酸轉(zhuǎn)氨酶(aspartate transaminase,AST)水平;肝臟組織石蠟切片蘇木精-伊紅(hematoxylin-eosin,HE)染色評(píng)價(jià)病理變化;末端脫氧核苷酸轉(zhuǎn)移酶介導(dǎo)d UTP缺口末端標(biāo)記測(cè)定法(Terminal deoxynucleoitidyl transferase-mediated d UTP nick end labeling,TUNEL)和Caspase-3活性測(cè)定評(píng)價(jià)凋亡情況;蛋白免疫印記法(western blot)和定量逆轉(zhuǎn)錄聚合酶鏈?zhǔn)椒磻?yīng)(quantitative reverse transcription polymerase chain reaction,q RT-PCR)檢測(cè)肝組織中細(xì)胞色P450 2E1(cytochrome P450 2E1,CYP 2E1)的表達(dá);谷胱甘肽(glutathione)的含量測(cè)定和過氧化氫酶(catalase,CAT)、超氧化物歧化酶(superoxide dismutase,SOD)的活性檢測(cè)評(píng)價(jià)肝臟的抗氧化能力;酶聯(lián)免疫吸附試驗(yàn)(enzyme-linked immunosorbent assay,ELISA)檢測(cè)血清中炎性介質(zhì)TNF-?和IL-1?的表達(dá);Western blot分析肝臟組織中Toll樣受體4(Toll-like receptor 4,TLR4)、p-IRAK1、p-I?B、p-p38蛋白表達(dá)水平。結(jié)果:1.阿魏酸呈劑量依賴性地降低了APAP誘導(dǎo)的ALT、AST水平。2.阿魏酸減輕了APAP誘導(dǎo)的肝臟病理改變和凋亡。HE染色結(jié)果顯示阿魏酸減少了APAP導(dǎo)致的壞死和出血程度。TUNEL染色和Caspase-3活性檢測(cè)結(jié)果表明了阿魏酸預(yù)處理后的細(xì)胞凋亡程度降低。3.阿魏酸顯著抑制了APAP誘導(dǎo)的CYP 2E1的m RNA和蛋白表達(dá)。4.阿魏酸明顯抑制了APAP誘導(dǎo)的氧化應(yīng)激反應(yīng)。阿魏酸干預(yù)后,肝組織GSH的含量顯著增多,SOD、CAT的活性顯著增強(qiáng)。5.阿魏酸顯著降低了APAP誘導(dǎo)的炎性因子TNF-?和IL-1?的表達(dá)。6.阿魏酸下調(diào)APAP誘導(dǎo)的TLR4、p-IRAK1、p-I?B、p-p38蛋白表達(dá)。結(jié)論:阿魏酸能減輕APAP誘導(dǎo)的肝毒性,這種保護(hù)作用可能與下調(diào)CYP 2E1表達(dá),負(fù)性調(diào)控TLR4信號(hào)通路,抑制炎癥反應(yīng)有關(guān)。
[Abstract]:Aim: to study the protective effect of Ferulic acidfon (FFA) on hepatic toxicity induced by acetaminophenophenol (APAP) and its mechanism. Methods: Balb/c mice were randomly divided into six groups: blank control group, ferulic acid control group, model group, low, medium and high dose ferulic acid intervention group. The model of acute liver injury was established by intraperitoneal injection of APAP(350 mg / kg in overnight fasting mice, the rats were given 10 ~ 30100 mg/kg of ferulic acid every 8 hours, and then intraperitoneally injected with ferulic acid as low, medium and high doses of ferulic acid. Ferulic acid (100 mg/kg) was used as the control group for 3 times, once every 8 h. Serum alanine aminotransferase (Alanine aminotransferase) and aspartate transaminase (aspartate transaminase) were detected after APAP18, and the pathological changes were evaluated by paraffin section of liver tissue. Terminal deoxynucleoitidyl transferase-mediated d UTP nick end labeling and Caspase-3 activity were used to evaluate the apoptosis of terminal deoxynucleoitidyl transferase-mediated d UTP nick end labelingn by terminal deoxynucleotidyl transferase mediated d UTP Nick end labeling assay. The expression of P450 2E1(cytochrome P450 2E1C CYP2E1) was detected by Western blot and quantitative reverse transcription polymerase chain reactionQ RT-PCRs in liver tissue by Western blot and quantitative reverse transcriptase polymerase chain reaction (RT-PCR). The determination of glutathione content and the activity of catalase CATX, superoxide dismutase (SOD) were used to evaluate the antioxidant ability of liver. The enzyme linked immunosorbent assay (Elisa) was used to detect the inflammatory mediators TNF-TNF in serum. And IL-1? The expression of Toll like receptor 4(Toll-like receptor 4 and TLR4 was analyzed by Western blot. The result is 1: 1. Ferulic acid decreased the level of alt induced by APAP in a dose-dependent manner. Ferulic acid alleviated the pathological changes and apoptosis induced by APAP. The results of HE staining showed that ferulic acid reduced the degree of necrosis and bleeding induced by APAP. Tunel staining and the detection of Caspase-3 activity showed that the apoptosis degree of the cells pretreated with ferulic acid was decreased by .3. Ferulic acid significantly inhibited the expression of m RNA and protein in CYP 2E1 induced by APAP. Ferulic acid significantly inhibited the oxidative stress induced by APAP. After ferulic acid intervention, the content of GSH in liver tissue increased significantly and the activity of cat in liver tissue increased significantly. Ferulic acid significantly reduced the inflammatory factor TNF-induced by APAP. And IL-1? 6. Ferulic acid down-regulated APAP induced expression of TLR4, p-IRAK1, p38. Conclusion: ferulic acid can attenuate the hepatotoxicity induced by APAP, which may be related to the down-regulation of CYP 2E1 expression, negative regulation of TLR4 signaling pathway and inhibition of inflammation.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R965

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 ;Protective effect of tea polyphenols against paracetamol-induced hepatotoxicity in mice is significanly correlated with cytochrome P450 suppression[J];World Journal of Gastroenterology;2009年15期

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本文編號(hào)：1874136