右美托咪定对梗阻性黄疸大鼠肝脏氧化应激损伤的影响

发布时间：2018-05-04 10:00

  本文选题：右美托咪定 + 氧化应激　； 参考：《大连医科大学》2017年硕士论文

【摘要】：目的:本文旨在研究右美托咪定对梗阻性黄疸大鼠肝脏氧化应激反应以及对HO-1蛋白表达的影响。方法:健康清洁级SD雄性大鼠40只,体重230～270g,采用随机数字表法,将大鼠分为4组,每组10只,依次分别为假手术组(S组)、梗阻性黄疸大鼠模型组(OJ组)、右美托咪定治疗组(D组,右美托咪定30ug/kg)、右美托咪定+Znpp组(D+Z组,右美托咪定30ug/kg,Znpp30mg/kg)。腹腔注射2%苯巴比妥钠溶液50mg/kg麻醉下,OJ组、D组和D+Z组行胆总管结扎(bileductligation,BDL),而S组大鼠不结扎胆总管。OJ组用生理盐水治疗,D组和D+Z组大鼠分别于取材前三天腹腔给予30ug/kg的右美托咪定注射液,连续给药三天,D+Z组在每次给予右美托咪定注射液半小时前,均腹腔注射30mg/kg的Znpp。在黄疸大鼠模型成立14天后检测并记录:1)心脏取血后采用全自动生化分析仪检测血清肝功能;2)采用ELISA法检测肝脏SOD活性、MDA含量;3)HE染色法观察肝脏病理组织学变化;4)采用免疫组化检测肝脏HO-1蛋白表达;5)Westen-blot检测HO-1蛋白水平的表达;6)Realtime-PCR 法检测 HO-lmRNA。结果:1.根据大鼠肝脏生化功能检测结果判断大鼠模型制作较成功。2.与S组相比,OJ组、D组和D+Z组血清ALT和AST水平显著增高,(p0.01);与OJ组和D+Z组相比,D组血清ALT和AST水平显著降低,其差异有统计学意义(p0.01);与OJ组相比,D+Z组血清AST和ALT差异无统计学意义(p0.05)。3.大鼠肝脏组织(MDA、SOD)的变化:在术后14天,与S组相比OJ组、D组、D+Z组的MDA含量显著增高,而SOD活性显著降低,其差异有统计学意义(p0.05);D组与OJ组和D+Z组比较MDA含量降低、SOD活性升高,差异有统计学意义(p0.05),D+Z组和OJ组相比MDA含量升高、SOD活性降低差异有意义(p0.05)。4.肝脏病理学观察:S组大鼠肝小叶结构正常,未见坏死肝细胞,肝细胞结构排列整齐;D组、D+Z组和OJ组可见肝脏组织不同程度的坏死;OJ组和D+Z组的大鼠肝组织出现严重的病理组织学损伤改变,包括明显的肝细胞变性,广泛但不同程度的肝细胞坏死,肝小叶结构被破坏、假小叶形成;D组大鼠肝组织损害明显改善,肝小叶分界较清晰,肝索排列相对整齐,绝大部分仅少量炎症细胞浸润。5.免疫组化检测肝脏HO-1蛋白表达:OJ组与S组相比大鼠肝脏内HO-1阳性染色结果明显加强,HO-1在肝内阳性结果染色颗粒及细胞数量明显增多,多种细胞内都有表达;与OJ组相比,D组显示大鼠肝脏HO-1染色阳性染色结果进一步加强,阳性染色结果的颗粒和细胞数量进一步增多;D+Z组与OJ组比较结果显示大鼠肝脏HO-1阳性结果细胞染色颗粒以及阳性面积未见明显差异。6.Western-blot方法显示,与S组相比,OJ组、D组和D+Z组HO-1蛋白表达增加(p0.05);与D组相比,D+Z组和OJ组Western-blot方法显示HO-1蛋白表达明显降低差异显著(p0.05);而D+Z组与OJ组比较HO-1蛋白表达无明显差异(p0.05)。7.Realtime-PCR方法检测大鼠肝脏HO-1mRNA:RT-PCR方法显示,与S组相比,OJ组HO-1m RNA表达增加(P0.05);与OJ组相比,D组HO-1mRNA表达进一步增加,差异有统计学意义(P0.05)。D+Z组与D组比较HO-1mRNA表达减少,差异有统计学意义(P0.05),但与OJ组比较差异无意义(P0.05)。结论:1.右美托咪定可通过抑制氧化应激反应减少梗阻性黄疸大鼠肝脏损伤。2.右美托咪定通过上调HO-1mRNA及其蛋白表达对梗阻性黄疸导致的肝损伤起保护作用。
[Abstract]:Objective: To study the effect of dexmedetomidin on oxidative stress and expression of HO-1 protein in the liver of rats with obstructive jaundice. Methods: 40 healthy and clean SD male rats, weighing 230 to 270g, were divided into 4 groups, 10 rats in each group, and the rats in each group were respectively the sham operation group (group S) and the rat model of obstructive jaundice, respectively. Group (group OJ), right metoimidin treatment group (group D, right metomimidine 30ug/kg), right metoimidine group +Znpp (D+Z group, right metomomidine 30ug/kg, Znpp30mg/kg). Under the abdominal injection of 2% phenobarbital solution 50mg/kg anesthesia, OJ group, D group and D+Z group were ligation of common bile duct (bileductligation,) in the group of rats without ligation of common bile duct. Treatment, group D and group D+Z rats were given 30ug/kg's right metoimidin injection three days before taking the material, and continuous administration for three days. Group D+Z was injected with right metoimidin for half an hour each time, 30mg/kg Znpp. was injected into the Jaundice Rat Model for 14 days and recorded: 1) after the heart was taken blood, automatic biochemical analysis was used. Test serum liver function; 2) ELISA assay was used to detect liver SOD activity, MDA content, 3) HE staining method to observe pathological changes of liver histopathology; 4) immunohistochemical detection of liver HO-1 protein expression; 5) Westen-blot detection of HO-1 protein level expression; 6) Realtime-PCR method to detect the result of HO-lmRNA.: 1. based on rat liver biochemical function test results Compared with group S, the serum levels of ALT and AST in group OJ, D and D+Z were significantly higher in group OJ, D and D+Z than in group OJ and D+Z group, and the difference of serum ALT and serum levels in D group was significantly lower than that in group OJ and D+Z group. The changes of A, SOD): 14 days after the operation, the MDA content in group OJ, D group and D+Z group was significantly higher than that of group S, while SOD activity decreased significantly, and the difference was statistically significant (P0.05); D group was lower than OJ group and D+Z group. P0.05.4. liver pathology observation: the rat liver lobule in group S was normal, no necrotic liver cells were found, and the liver cell structure was neatly arranged. Group D, group D+Z and OJ group showed necrosis of liver tissue in different degrees, and serious pathological changes of pathological histology in the liver tissue of group OJ and D+Z group, including obvious liver cell degeneration, extensive but not. The liver necrosis of the same degree, the destruction of hepatic lobule structure and the formation of false lobule, the damage of liver tissue in the D group was obviously improved, the demarcation of hepatic lobule was clear, the arrangement of hepatic cord was relatively neat, and the vast majority of only a small amount of inflammatory cells infiltrated.5. immunohistochemistry to detect the expression of HO-1 protein in the liver, and the results of HO-1 positive staining in the liver of the group of the rats were compared with that of the S group. The positive results of the positive staining particles and cell number of HO-1 in the liver were significantly increased and the number of cells in all kinds of cells were expressed. Compared with the OJ group, the D group showed that the positive staining results of HO-1 staining in the liver of rats were further strengthened, and the number of particles and cells in the positive staining results increased further; the D +Z group compared with the OJ group, the results showed the positive liver HO-1 in the rat liver. Compared with the S group, the expression of HO-1 protein in OJ group, D group and D+Z group increased (P0.05) compared with the group of OJ, and the Western-blot method of D+Z group and OJ group showed significant difference in the expression of HO-1 protein expression compared with that of the group D, but there was no significant difference in the expression of the protein expression in the group of D+Z and OJ groups. Compared with the S group, the expression of HO-1m RNA in the OJ group increased (P0.05), compared with the group S, and the HO-1mRNA expression of the D group was further increased, compared with the group of OJ (P0.05), and the difference has statistical significance compared with that of the OJ group (P0.05), the difference has statistical significance, but the difference is worse than that of the group. P0.05. Conclusion: 1. dexmedetomidine can protect the liver injury caused by obstructive jaundice by inhibiting the oxidative stress reaction to reduce the liver damage of the obstructive jaundice rats.2. right metomimidine by up regulation of HO-1mRNA and its protein expression.

【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R614
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本文编号：1842603