發(fā)布時間：2018-05-31 06:15

  本文選題：梔子提取物 + 酒精性肝損害��； 參考：《遵義醫(yī)學院》2014年碩士論文

【摘要】：目的：構建SD大鼠酒精性肝病的動物模型，并灌胃梔子提取物進行干預，然后通過測試實驗大鼠的血清脂質總膽固醇(TC)、甘油三酯(TG)、谷丙氨酸氨基轉移酶(ALT)、天冬氨酸氨基轉移酶(AST)的水平，測定肝臟指數及肝臟組織中谷胱甘肽過氧化物酶(GSH-Px)、丙二醛(MDA)的含量，再觀測肝臟組織腫瘤壞死因子-α (TNF-α)、白細胞介素-6(IL-6)蛋白表達和基因轉錄水平以及大鼠肝細胞脂肪變情況，以便探研酒精性肝病的病理機制及梔子提取物對其病變的影響。 方法：五十只健康的SD雄性大鼠，隨機分為五組（n=10），正常對照組（對照組）、酒精性肝病模型組（模型組）、梔子提取物低劑量組（低劑量組）、梔子提取物中劑量組（中劑量組）和梔子提取物高劑量組（高劑量組）。除了對照組大鼠給予普通飼料和清潔飲用水外，其余各組均每天給予高脂飼料和56%乙醇15ml/kg每天灌胃，各藥物劑量組同時每天再添加低、中、高劑量的梔子提取物灌胃，，共連續(xù)飼養(yǎng)8周。然后分別麻醉動物，取材制備實驗動物標本。（1）采取肝臟用電子天平稱重，計算肝指數；（2）采用生化法檢測血清的TC、TG、ALT和AST，并用試劑盒檢測大鼠肝臟中GSH-Px和MDA的含量；（3）通過HE染色、免疫組化染色法分別用光鏡觀察肝組織的形態(tài)變化和TNF-α、IL-6的表達變化陽性指標；（4）提取肝臟組織的RNA，通過熒光定量RT-PCR法分別檢測肝組織中TNF-α和IL-6mRNA的基因轉錄水平。 結果：（1）動物用藥8周后，計算大鼠肝指數，發(fā)現(xiàn)模型組較對照組的肝指數增高（P＜0.05），各藥物組肝指數較模型組則明顯降低（P＜0.05）。（2）模型組大鼠血清的TC、TG含量明顯高于對照組（P＜0.05），各藥物組與模型組比較，血清脂質明顯降低（P＜0.05），模型組肝功能ALT、AST值明顯較高于對照組（P＜0.05），各藥物組肝功能指標則低于模型組（P＜0.05）；肝臟MDA含量模型組明顯高于對照組（P＜0.05），各藥物組MDA含量較模型組顯著降低（P＜0.05）；模型組GSH-Px含量明顯低于對照組（P＜0.05），藥物組GSH-Px含量明顯比模型組提高（P＜0.05）。（3）HE染色光鏡下觀察，各藥物組的肝細胞脂肪變程度相比模型組大鼠而言有明顯改善；免疫組化染色結果顯示模型組較對照組大鼠肝臟組織的TNF-α、IL-6表達增多（P＜0.05），各藥物組明顯比模型組降低（P＜0.05）。（4）RT-PCR結果表明，模型組肝組織內TNF-α、IL-6mRNA的基因表達量均明顯高于對照組，也高于各藥物組（P＜0.05）。 結論：（1）成功建立了SD大鼠酒精性肝臟損害的模型，并應用了梔子提取物來干預改善模型組大鼠的肝損害。（2）模型組大鼠的血清TC、TG、ALT、AST水平和MDA含量升高，GSH-Px含量下降；利用梔子提取物來進行藥物干預，可以使血清TC、TG、ALT、AST水平和MDA含量降低，GSH-Px含量升高。（3）模型組大鼠肝組織的TNF-α、IL-6基因轉錄和蛋白表達升高，采用梔子提取物的藥物干預可使其降低。 總的來看，模型組SD大鼠的血清脂質的TC和TG水平升高,肝臟的ALT、AST和MDA水平升高，而GSH-Px含量降低，以及肝組織的TNF-α和IL-6基因轉錄表達增高，與酒精性肝臟損害有關；梔子提取物可能是通過下調TNF-α和IL-6轉錄表達、促進脂肪代謝、清除肝組織內自由基等，從而發(fā)揮保肝的治療作用。
[Abstract]:Objective: to construct an animal model of alcoholic liver disease in SD rats, and to interfere with the extract of Gardenia jasminoides gardenia, and then test the level of serum lipid total cholesterol (TC), triglyceride (TG), alanine aminotransferase (ALT) and aspartate aminotransferase (AST), and determine the liver index and glutathione peroxy in the liver tissue. The content of chemical enzyme (GSH-Px) and malondialdehyde (MDA), the expression of tumor necrosis factor - alpha (TNF- alpha), the expression of interleukin -6 (IL-6) protein and gene transcription level, and the fatty change of rat liver cells, in order to explore the pathological mechanism of alcoholic liver disease and the effect of the extract of Gardenia jasminoides on its pathological changes.
Methods: Fifty healthy SD male rats were randomly divided into five groups (n=10), normal control group (control group), alcoholic liver disease model group (model group), low dose group of Gardenia extract (low dose group), middle dose group of Gardenia extract (medium dose group) and high dose group of Gardenia extract (high dose group). Except for control group, rats were given ordinary diet. And the other groups were given high fat diet and 56% ethanol 15ml/kg every day. Each drug dose group added low, medium and high doses of Gardenia extract for 8 weeks. Then the animals were reared for 8 weeks. Then, the animals were anaesthetized and prepared to prepare the experimental animal specimens. (1) the liver scales were weighed and the hepatic finger was calculated. (2) TC, TG, ALT and AST were detected by biochemical method, and the content of GSH-Px and MDA in rat liver was detected by the kit. (3) the morphological changes of liver tissue and the positive index of TNF- A and IL-6 expression were observed by HE staining and immunohistochemical staining method respectively. (4) the RNA of the liver tissue was extracted by the fluorescent quantitative RT-PCR method. The transcriptional levels of TNF- and IL-6mRNA genes were detected in liver tissues.
Results: (1) after 8 weeks of drug use, the liver index of rats was calculated and the liver index of the model group was higher than that of the control group (P < 0.05). The liver index of each group was significantly lower than that in the model group (P < 0.05). (2) the serum TC, TG content of the model group was significantly higher than that of the control group (P < 0.05). The serum lipids were significantly lower than those in the model group (P < < < < 0. 0.05) in the model group, the liver function ALT, AST value was significantly higher than the control group (P < 0.05), the liver function index of each drug group was lower than that of the model group (P < 0.05), the liver MDA content model group was significantly higher than the control group (P < 0.05), the MDA content in each drug group was significantly lower than the model group (P < 0.05), and the GSH-Px content in the model group was significantly lower than that of the control group (P < 0.05). The drug was significantly lower than the control group (P < 0.05). The content of GSH-Px in the group was significantly higher than that in the model group (P < 0.05). (3) under the HE staining light microscope, the fatty change degree of the liver cells in each drug group was significantly improved compared with the model group. The immunohistochemical staining results showed that the expression of TNF- a and IL-6 in the liver tissue of the model group increased (P < 0.05), and the drug groups were obviously compared with the model group. The group decreased (P < 0.05). (4) the results of RT-PCR showed that the gene expression of TNF- alpha and IL-6mRNA in the liver tissue of the model group was significantly higher than that in the control group, and was higher than that of the drug groups (P < 0.05).
Conclusion: (1) the model of alcoholic liver damage in SD rats was successfully established and the extract of Gardenia jasminoides was used to intervene to improve the liver damage in the model rats. (2) the serum levels of TC, TG, ALT, AST and MDA in the model group were increased and the content of GSH-Px decreased, and the serum TC, TG, ALT and AST levels could be made by using the extract of Gardenia jasminoides. And the content of MDA decreased and the content of GSH-Px increased. (3) the transcriptional and protein expression of TNF- alpha and IL-6 gene in the rat liver tissue of the model group increased, and the drug intervention by the extract of Gardenia could reduce it.
In total, the level of serum lipid TC and TG in the model group SD rats increased, the level of ALT, AST and MDA in the liver increased, but the content of GSH-Px decreased, and the transcriptional expression of TNF- A and IL-6 genes in the liver tissues increased, which was related to the alcoholic liver damage. The extract of Gardenia may be through down regulation of TNF- A and IL-6 transcription, promoting fat metabolism, and In addition to the free radicals in liver tissue, the therapeutic effect of liver protection is realized.
【學位授予單位】：遵義醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R285.5

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