右美托咪定对肾缺血再灌注后肝损伤的保护作用及机制研究

发布时间：2018-05-26 17:40

本文选题：缺血再灌注 + 肝损伤　；参考：《锦州医科大学》2017年硕士论文

【摘要】：目的探讨不同剂量右美托咪定(DEX)预处理对小鼠肾脏IRI导致肝损伤的保护作用及机制。方法动物实验:40只8周龄健康雄性C57 BL/6J小鼠,随机均分为5组:sham组、IRI组、25μg/kg Dex组(低剂量)、50μg/kg Dex组(中剂量)和100μg/kg Dex组(高剂量),每组8只。除sham组外,其余各组小鼠均在背部中线两侧肾区切口,用玻璃分针钝性分离肾脏周围肌肉组织和肾筋膜,用无创伤动脉夹轻轻夹闭双侧肾脏动脉血管导致肾缺血,进行造模。于手术后24h处死小鼠,收集血液、肾脏和肝脏组织。检测小鼠肾功能和肝功能指标,HE染色观察小鼠肾脏和肝脏病理改变,WST-1法检测小鼠肝脏组织总超氧化物歧化酶(SOD)活力,TBA法测定小鼠肝脏组织丙二醛(MDA)含量,比色法测定小鼠肾脏组织谷胱甘肽过氧化物酶(GSH-PX)含量,ELISA法检测了小鼠肝脏匀浆液内炎症因子TNF-α、MCP-1、和IL-6以及抗炎症因子IL-10含量,Western-blot方法检测分析小鼠肝脏组织Caspase3、Bcl-2和Bax蛋白表达情况。结果1.Dex预处理后可以明显改善由于肾脏IRI导致的肾功能改变:与sham相比较,IR组小鼠血浆中Scr,Bun和Cys-C显著升高(P0.01);与IR组相比较,25ug/kg Dex组小鼠血浆中Bun下降不明显(P0.05),Scr和Cys-C明显降低(P0.05);50 ug/kg、100 ug/kg Dex组小鼠血浆中Scr,Bun和Cys-C明显降低(P0.05)。2.Dex预处理后可以明显改善由于肾脏IRI导致的肝功能改变:与sham相比较,IR组小鼠血浆中AST和ALT的含量均显著升高(P0.01);与IR组相比较,25ug/kg Dex、50 ug/kg、100 ug/kg Dex组小鼠血浆中AST和ALT含量均下降比较明显(P0.05)。3.Dex预处理后可以明显改善由于肾脏IRI导致的肾脏结构病变情况:与sham相比较,IR组小鼠病变明显,HE染色显示肾脏皮质和髓质交界处肾小管细胞坏死脱落;与IR组相比较,25ug/kg Dex、50 ug/kg Dex、100 ug/kg Dex组小鼠肾脏病变明显得到改善。4.Dex预处理后可以明显改善由于肾脏IRI导致的肝脏结构病变情况:与sham相比较,IR组小鼠病变明显,HE染色显示IR组小鼠肝脏大量炎性细胞浸润,肝窦扩张,少量空泡形成,肝细胞排列紊乱。与IR组相比较,25ug/kg Dex、50 ug/kg Dex、100 ug/kg Dex组小鼠肝脏病变明显得到改善。5.Dex预处理后可以明显提高肝脏对氧自由基的清除能力,提高组织内总抗氧化能力:与sham相比较,IR组小鼠肝脏中SOD活力和GSH-PX活性明显降低(P0.05),MDA含量显著升高(P0.01);与IR组相比较,25ug/kg Dex、50 ug/kg DEX、100 ug/kg Dex组小鼠肝脏中SOD活力和GSH-PX活性明显升高(P0.05),MDA含量显著降低(P0.05)。6.Dex预处理后可以明显改善肝脏组织的抗炎症能力:与sham相比较,IR组小鼠肝脏中TNF-α、IL-6、IL-10和MCP-1含量均升高(P0.05);与IR组相比较,25ug/kg Dex、50 ug/kg、100 ug/kg Dex预处理组小鼠肝脏TNF-α、MCP-1和IL-6含量明显下降(P0.05),IL-10含量升高明显(P0.01)。7.Dex预处理后可以明显改善肝脏组织的细胞凋亡情况:与sham相比较,IR组小鼠肝脏中Bcl-2蛋白表达量下降(P0.01),Bax和Caspase 3蛋白表达量升高(P0.01);与IR组相比较,25ug/kg Dex组小鼠肝脏中Bax蛋白表达量下降(P0.05),Caspase 3和Bcl-2蛋白表达下降不明显(P0.05);50 ug/kg DEX、100 ug/kg Dex组小鼠肝脏中Bcl-2蛋白表达量升高(P0.05),Bax和Caspase 3蛋白表达量下降(P0.05)。结论1.肾脏缺血再灌注可导致肝脏损伤。2.DEX对肾脏IR导致肝脏损伤的保护作用可以通过提高肝脏抗氧化和抗凋亡能力来发挥的。3.50 ug/kg、100 ug/kg Dex预处理对肾脏IR导致的肝脏损伤的保护作用比较明显。
[Abstract]:Objective to investigate the protective effect and mechanism of different doses of dexmedetomidine (DEX) preconditioning on liver injury induced by IRI in mice kidney. Method animal experiment: 40 healthy male C57 BL/6J mice of 8 weeks old were randomly divided into 5 groups: sham group, IRI group, 25 mu g/kg Dex group (low dose), 50 micron Dex group (middle dose) and 100 mu g/kg Dex group (Gao Jiliang), 8 rats in each group, 8 rats in each group. In addition to group sham, all the mice in the other groups were incisive in the bilateral renal region of the middle back of the back, with a blunt separation of the surrounding muscle tissue and the fascia of the kidney with a glass needle. The renal ischemia was caused by the unwound artery clamp and the renal ischemia was created by the unwound artery clamp. The mice were killed and the kidney and liver tissues were collected after the operation. The kidney and the liver tissues were collected and the kidney was collected to detect the kidney of the mice. The pathological changes of kidney and liver in mice were observed by HE staining. The activity of total superoxide dismutase (SOD) in liver tissues of mice was detected by WST-1. The content of malondialdehyde (MDA) in liver tissue of mice was measured by TBA, and the content of glutathione peroxidase (GSH-PX) in kidney tissue of mice was measured by colorimetry. The liver homogenization of mice was detected by ELISA method. The content of inflammatory factors TNF- alpha, MCP-1, IL-6, and anti-inflammatory factor IL-10 in the serous, and the Western-blot method to detect the expression of Caspase3, Bcl-2 and Bax protein in the liver tissues of mice. Results the renal function changes caused by IRI were obviously improved by 1.Dex pretreatment. Compared with group IR, the decrease of Bun in the plasma of 25ug/kg Dex group was not obvious (P0.05), and Scr and Cys-C decreased significantly (P0.05); 50 ug/kg, 100 ug/kg Dex group mice plasma The content of AST and ALT in plasma increased significantly (P0.01). Compared with the IR group, the levels of AST and ALT in the plasma of 25ug/kg Dex, 50 ug/kg and 100 ug/kg Dex mice were significantly decreased. Compared with the IR group, the renal lesions of the 25ug/kg Dex, 50 ug/kg Dex, and the 100 ug/kg Dex group were obviously improved by the improvement of.4.Dex preconditioning, and the pathological changes in the kidney caused by IRI were obviously improved. Compared with the sham phase, the IR group had obvious lesions, HE staining showed A large number of inflammatory cells in the liver of IR mice were infiltrated, the hepatic sinusoids dilated, a small amount of vacuoles formed and the liver cells were arranged in disorder. Compared with the IR group, 25ug/kg Dex, 50 ug/kg Dex, and the liver lesions of the 100 ug/kg Dex mice were obviously improved by the improvement of.5.Dex preconditioning, and the total antioxidant capacity in the liver was improved and the total antioxidant capacity in the tissues was improved. Compared with sham, the activity of SOD and the activity of GSH-PX in the liver of IR mice decreased significantly (P0.05), and the content of MDA increased significantly (P0.01). Compared with the IR group, 25ug/kg Dex, 50 ug/kg DEX, the activity and activity of the liver in the liver of the 100 mice were significantly increased. Compared with sham, the levels of TNF- alpha, IL-6, IL-10 and MCP-1 in the liver of IR mice increased (P0.05), and compared with those in the IR group, 25ug/kg Dex, 50 ug/kg, and the 100 ug/kg pretreated mice were significantly improved. The apoptosis of liver tissue: compared with sham, the expression of Bcl-2 protein in the liver of IR mice decreased (P0.01), and the expression of Bax and Caspase 3 protein increased (P0.01). Compared with the IR group, the expression of Bax protein in the liver of the 25ug/kg Dex group decreased (P0.05), and the decline of the 3 and the protein expression was not obvious; 50 The expression of Bcl-2 protein in the liver of /kg Dex mice increased (P0.05) and the expression of Bax and Caspase 3 protein decreased (P0.05). Conclusion 1. renal ischemia-reperfusion can lead to the protective effect of liver injury.2.DEX on kidney IR induced liver damage, which can be achieved by improving the liver antioxidant and anti withering ability of.3.50 ug/kg, 100 ug/kg. The protective effect of Li on liver injury caused by IR is obvious.
【学位授予单位】：锦州医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R614

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