Ghrelin對酒精性肝損傷的保護(hù)作用及其機(jī)制研究
發(fā)布時間：2018-12-11 18:30
【摘要】：目的: Ghrelin是生長激素釋放促分泌素受體的一種內(nèi)源性配體,最近研究表明其對非酒精脂肪肝具有保護(hù)作用,基于以上研究基礎(chǔ)，我們采用了乙醇誘導(dǎo)小鼠酒精性肝損傷模型，研究不同劑量的Ghrelin對酒精性肝損傷小鼠體內(nèi)轉(zhuǎn)氨酶（ALT、AST）、氧自由基（MDA、SOD、GSH-Px）、炎性因子（TNF-α、IL-1β、IFN-γ和MCP-1）及肝臟病理學(xué)的影響。 研究方法：設(shè)正常對照組、模型組、Ghrelin給藥組(5，10，20μg·kg-1)。將動物按體重分層隨機(jī)分組，每組6只。模型組及Ghrelin給藥組給予含5%乙醇的液體飼料，自由喂養(yǎng)4周，最后一次按5g·kg-1的劑量給予酒精灌胃一次，建立酒精性肝損傷模型。給藥組每天向小鼠腹腔內(nèi)注射濃度分別為5，10，20μg·kg-1的Ghrelin。正常組及模型組予以等量的生理鹽水。酒精灌胃9小時后自小鼠后眼眶靜脈叢采血，拉頸處死動物，立即摘取肝臟標(biāo)本進(jìn)行有關(guān)檢測。在誘導(dǎo)酒精性肝損傷模型的基礎(chǔ)上，用分光光度法檢測血清中ALT、AST和肝勻漿MDA、SOD、GSH-Px含量；Real-time PCR法檢測肝臟中細(xì)胞因子TNF-α、IL-1β等的表達(dá)。HE染色檢測肝臟病理改變。 結(jié)果:在序貫給予酒精飼料4周后，模型組血漿ALT、AST水平升高，而在序貫期間預(yù)防給藥，分別給予ip Ghrelin(5，10，20μg·kg-1)均能降低血漿中升高的ALT、AST水平，在Ghrelin的3個劑量中以20μg·kg-1為最明顯; HE染色發(fā)現(xiàn)，模型組肝內(nèi)出現(xiàn)彌漫性脂肪病變，肝血竇充血明顯，有少許炎細(xì)胞浸潤。Ghrelin處理可減輕炎癥細(xì)胞浸潤，但對脂肪病變影響不明顯;模型組肝勻漿MDA水平升高，GSH-Px和SOD活性降低。預(yù)防給予ip Ghrelin(5，10，20μg·kg-1)均能降低肝勻漿中升高M(jìn)DA的水平，并能提高肝勻漿GSH-Px和SOD水平;與正常組相比，模型組肝臟炎癥細(xì)胞因子如TNF-α、IL-1β、IFN-和MCP-1明顯升高。預(yù)防給予ip Ghrelin(5，10，20μg·kg-1)均能降低上述炎性細(xì)胞因子的mRNA表達(dá)。 結(jié)論：本研究首次在酒精性肝損傷動物模型上發(fā)現(xiàn)Ghrelin的抗炎作用，，Ghrelin可抑制酒精引起的肝臟炎癥細(xì)胞因子的表達(dá)。而Ghrelin通過何種機(jī)制抑制炎性細(xì)胞因子的表達(dá)值得進(jìn)一步研究。
[Abstract]:Objective: Ghrelin is an endogenous ligand of growth hormone releasing secretin receptor. Recent studies have shown that it has protective effect on non-alcoholic fatty liver. Based on the above research, we used ethanol to induce alcoholic liver injury in mice. To study the effects of different doses of Ghrelin on transaminase (ALT,AST), oxygen free radical (MDA,SOD,GSH-Px), inflammatory factors (TNF- 偽, IL-1 尾, IFN- 緯 and MCP-1) and liver pathology in mice with alcoholic liver injury. Methods: normal control group, model group and Ghrelin administration group (5 ~ 1020 渭 g kg-1) were set up. Animals were randomly divided into 6 groups according to their body weight. The model group and the Ghrelin group were given 5% ethanol as a liquid diet. The rats were fed freely for 4 weeks, and the last dose of 5 g kg-1 was used to establish the model of alcoholic liver injury. The mice in the administration group were injected intraperitoneally with Ghrelin. of 20 渭 g / kg-1 at the concentration of 5 渭 g / min each day. The normal group and the model group were given the same amount of normal saline. The blood was collected from the orbital venous plexus of the mice after 9 hours of gastric perfusion, and the animals were killed by pulling the neck, and the liver samples were removed immediately for detection. Based on the model of alcoholic liver injury, serum ALT,AST and liver homogenate MDA,SOD,GSH-Px were determined by spectrophotometry. The expressions of cytokines TNF- 偽 and IL-1 尾 in liver were detected by Real-time PCR method. The pathological changes of liver were detected by HE staining. Results: after 4 weeks of sequential administration of alcohol feed, the plasma ALT,AST level in the model group was increased, while during the sequential administration, the plasma ALT,AST level was significantly decreased by ip Ghrelin (10 ~ 10 渭 g / kg-1). Among the three doses of Ghrelin, 20 渭 g kg-1 was the most obvious. HE staining showed that there were diffuse fatty lesions in the liver of the model group, the hepatic sinusoidal congestion was obvious, and a few inflammatory cells were infiltrated. Ghrelin treatment could reduce the inflammatory cell infiltration, but had no obvious effect on the fatty lesions. In the model group, the level of MDA in liver homogenate increased and the activities of GSH-Px and SOD decreased. Prophylactic administration of ip Ghrelin (10 渭 g kg-1) decreased the level of MDA in liver homogenate, and increased the levels of GSH-Px and SOD in liver homogenate. Compared with the normal group, the inflammatory cytokines such as TNF- 偽, IL-1 尾, IFN- and MCP-1 increased significantly in the model group. Prophylaxis of ip Ghrelin (5 (10 ~ 10 渭 g kg-1) decreased the mRNA expression of above inflammatory cytokines. Conclusion: in this study, the anti-inflammatory effect of Ghrelin was first found in alcoholic liver injury animal model. Ghrelin can inhibit the expression of inflammatory cytokines in the liver induced by alcohol. The mechanism by which Ghrelin inhibits the expression of inflammatory cytokines deserves further study.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R575.5
本文編號：2373021
[Abstract]:Objective: Ghrelin is an endogenous ligand of growth hormone releasing secretin receptor. Recent studies have shown that it has protective effect on non-alcoholic fatty liver. Based on the above research, we used ethanol to induce alcoholic liver injury in mice. To study the effects of different doses of Ghrelin on transaminase (ALT,AST), oxygen free radical (MDA,SOD,GSH-Px), inflammatory factors (TNF- 偽, IL-1 尾, IFN- 緯 and MCP-1) and liver pathology in mice with alcoholic liver injury. Methods: normal control group, model group and Ghrelin administration group (5 ~ 1020 渭 g kg-1) were set up. Animals were randomly divided into 6 groups according to their body weight. The model group and the Ghrelin group were given 5% ethanol as a liquid diet. The rats were fed freely for 4 weeks, and the last dose of 5 g kg-1 was used to establish the model of alcoholic liver injury. The mice in the administration group were injected intraperitoneally with Ghrelin. of 20 渭 g / kg-1 at the concentration of 5 渭 g / min each day. The normal group and the model group were given the same amount of normal saline. The blood was collected from the orbital venous plexus of the mice after 9 hours of gastric perfusion, and the animals were killed by pulling the neck, and the liver samples were removed immediately for detection. Based on the model of alcoholic liver injury, serum ALT,AST and liver homogenate MDA,SOD,GSH-Px were determined by spectrophotometry. The expressions of cytokines TNF- 偽 and IL-1 尾 in liver were detected by Real-time PCR method. The pathological changes of liver were detected by HE staining. Results: after 4 weeks of sequential administration of alcohol feed, the plasma ALT,AST level in the model group was increased, while during the sequential administration, the plasma ALT,AST level was significantly decreased by ip Ghrelin (10 ~ 10 渭 g / kg-1). Among the three doses of Ghrelin, 20 渭 g kg-1 was the most obvious. HE staining showed that there were diffuse fatty lesions in the liver of the model group, the hepatic sinusoidal congestion was obvious, and a few inflammatory cells were infiltrated. Ghrelin treatment could reduce the inflammatory cell infiltration, but had no obvious effect on the fatty lesions. In the model group, the level of MDA in liver homogenate increased and the activities of GSH-Px and SOD decreased. Prophylactic administration of ip Ghrelin (10 渭 g kg-1) decreased the level of MDA in liver homogenate, and increased the levels of GSH-Px and SOD in liver homogenate. Compared with the normal group, the inflammatory cytokines such as TNF- 偽, IL-1 尾, IFN- and MCP-1 increased significantly in the model group. Prophylaxis of ip Ghrelin (5 (10 ~ 10 渭 g kg-1) decreased the mRNA expression of above inflammatory cytokines. Conclusion: in this study, the anti-inflammatory effect of Ghrelin was first found in alcoholic liver injury animal model. Ghrelin can inhibit the expression of inflammatory cytokines in the liver induced by alcohol. The mechanism by which Ghrelin inhibits the expression of inflammatory cytokines deserves further study.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R575.5
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本文編號：2373021
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