Ghrelin对酒精性肝损伤的保护作用及其机制研究
发布时间：2018-12-11 18:30
【摘要】：目的: Ghrelin是生长激素释放促分泌素受体的一种内源性配体,最近研究表明其对非酒精脂肪肝具有保护作用,基于以上研究基础，我们采用了乙醇诱导小鼠酒精性肝损伤模型，研究不同剂量的Ghrelin对酒精性肝损伤小鼠体内转氨酶（ALT、AST）、氧自由基（MDA、SOD、GSH-Px）、炎性因子（TNF-α、IL-1β、IFN-γ和MCP-1）及肝脏病理学的影响。 研究方法：设正常对照组、模型组、Ghrelin给药组(5，10，20μg·kg-1)。将动物按体重分层随机分组，每组6只。模型组及Ghrelin给药组给予含5%乙醇的液体饲料，自由喂养4周，最后一次按5g·kg-1的剂量给予酒精灌胃一次，建立酒精性肝损伤模型。给药组每天向小鼠腹腔内注射浓度分别为5，10，20μg·kg-1的Ghrelin。正常组及模型组予以等量的生理盐水。酒精灌胃9小时后自小鼠后眼眶静脉丛采血，拉颈处死动物，立即摘取肝脏标本进行有关检测。在诱导酒精性肝损伤模型的基础上，用分光光度法检测血清中ALT、AST和肝匀浆MDA、SOD、GSH-Px含量；Real-time PCR法检测肝脏中细胞因子TNF-α、IL-1β等的表达。HE染色检测肝脏病理改变。 结果:在序贯给予酒精饲料4周后，模型组血浆ALT、AST水平升高，而在序贯期间预防给药，分别给予ip Ghrelin(5，10，20μg·kg-1)均能降低血浆中升高的ALT、AST水平，在Ghrelin的3个剂量中以20μg·kg-1为最明显; HE染色发现，模型组肝内出现弥漫性脂肪病变，肝血窦充血明显，有少许炎细胞浸润。Ghrelin处理可减轻炎症细胞浸润，但对脂肪病变影响不明显;模型组肝匀浆MDA水平升高，GSH-Px和SOD活性降低。预防给予ip Ghrelin(5，10，20μg·kg-1)均能降低肝匀浆中升高MDA的水平，并能提高肝匀浆GSH-Px和SOD水平;与正常组相比，模型组肝脏炎症细胞因子如TNF-α、IL-1β、IFN-和MCP-1明显升高。预防给予ip Ghrelin(5，10，20μg·kg-1)均能降低上述炎性细胞因子的mRNA表达。 结论：本研究首次在酒精性肝损伤动物模型上发现Ghrelin的抗炎作用，，Ghrelin可抑制酒精引起的肝脏炎症细胞因子的表达。而Ghrelin通过何种机制抑制炎性细胞因子的表达值得进一步研究。
[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- 伪, IL-1 尾, IFN- 纬 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- 伪 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- 伪, 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.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R575.5
本文编号：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- 伪, IL-1 尾, IFN- 纬 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- 伪 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- 伪, 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.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R575.5
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