本文選題：右美托咪定 + 氧化應(yīng)激�。� 參考：《大連醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:本文旨在研究右美托咪定對(duì)梗阻性黃疸大鼠肝臟氧化應(yīng)激反應(yīng)以及對(duì)HO-1蛋白表達(dá)的影響。方法:健康清潔級(jí)SD雄性大鼠40只,體重230～270g,采用隨機(jī)數(shù)字表法,將大鼠分為4組,每組10只,依次分別為假手術(shù)組(S組)、梗阻性黃疸大鼠模型組(OJ組)、右美托咪定治療組(D組,右美托咪定30ug/kg)、右美托咪定+Znpp組(D+Z組,右美托咪定30ug/kg,Znpp30mg/kg)。腹腔注射2%苯巴比妥鈉溶液50mg/kg麻醉下,OJ組、D組和D+Z組行膽總管結(jié)扎(bileductligation,BDL),而S組大鼠不結(jié)扎膽總管。OJ組用生理鹽水治療,D組和D+Z組大鼠分別于取材前三天腹腔給予30ug/kg的右美托咪定注射液,連續(xù)給藥三天,D+Z組在每次給予右美托咪定注射液半小時(shí)前,均腹腔注射30mg/kg的Znpp。在黃疸大鼠模型成立14天后檢測(cè)并記錄:1)心臟取血后采用全自動(dòng)生化分析儀檢測(cè)血清肝功能;2)采用ELISA法檢測(cè)肝臟SOD活性、MDA含量;3)HE染色法觀察肝臟病理組織學(xué)變化;4)采用免疫組化檢測(cè)肝臟HO-1蛋白表達(dá);5)Westen-blot檢測(cè)HO-1蛋白水平的表達(dá);6)Realtime-PCR 法檢測(cè) HO-lmRNA。結(jié)果:1.根據(jù)大鼠肝臟生化功能檢測(cè)結(jié)果判斷大鼠模型制作較成功。2.與S組相比,OJ組、D組和D+Z組血清ALT和AST水平顯著增高,(p0.01);與OJ組和D+Z組相比,D組血清ALT和AST水平顯著降低,其差異有統(tǒng)計(jì)學(xué)意義(p0.01);與OJ組相比,D+Z組血清AST和ALT差異無(wú)統(tǒng)計(jì)學(xué)意義(p0.05)。3.大鼠肝臟組織(MDA、SOD)的變化:在術(shù)后14天,與S組相比OJ組、D組、D+Z組的MDA含量顯著增高,而SOD活性顯著降低,其差異有統(tǒng)計(jì)學(xué)意義(p0.05);D組與OJ組和D+Z組比較MDA含量降低、SOD活性升高,差異有統(tǒng)計(jì)學(xué)意義(p0.05),D+Z組和OJ組相比MDA含量升高、SOD活性降低差異有意義(p0.05)。4.肝臟病理學(xué)觀察:S組大鼠肝小葉結(jié)構(gòu)正常,未見(jiàn)壞死肝細(xì)胞,肝細(xì)胞結(jié)構(gòu)排列整齊;D組、D+Z組和OJ組可見(jiàn)肝臟組織不同程度的壞死;OJ組和D+Z組的大鼠肝組織出現(xiàn)嚴(yán)重的病理組織學(xué)損傷改變,包括明顯的肝細(xì)胞變性,廣泛但不同程度的肝細(xì)胞壞死,肝小葉結(jié)構(gòu)被破壞、假小葉形成;D組大鼠肝組織損害明顯改善,肝小葉分界較清晰,肝索排列相對(duì)整齊,絕大部分僅少量炎癥細(xì)胞浸潤(rùn)。5.免疫組化檢測(cè)肝臟HO-1蛋白表達(dá):OJ組與S組相比大鼠肝臟內(nèi)HO-1陽(yáng)性染色結(jié)果明顯加強(qiáng),HO-1在肝內(nèi)陽(yáng)性結(jié)果染色顆粒及細(xì)胞數(shù)量明顯增多,多種細(xì)胞內(nèi)都有表達(dá);與OJ組相比,D組顯示大鼠肝臟HO-1染色陽(yáng)性染色結(jié)果進(jìn)一步加強(qiáng),陽(yáng)性染色結(jié)果的顆粒和細(xì)胞數(shù)量進(jìn)一步增多;D+Z組與OJ組比較結(jié)果顯示大鼠肝臟HO-1陽(yáng)性結(jié)果細(xì)胞染色顆粒以及陽(yáng)性面積未見(jiàn)明顯差異。6.Western-blot方法顯示,與S組相比,OJ組、D組和D+Z組HO-1蛋白表達(dá)增加(p0.05);與D組相比,D+Z組和OJ組Western-blot方法顯示HO-1蛋白表達(dá)明顯降低差異顯著(p0.05);而D+Z組與OJ組比較HO-1蛋白表達(dá)無(wú)明顯差異(p0.05)。7.Realtime-PCR方法檢測(cè)大鼠肝臟HO-1mRNA:RT-PCR方法顯示,與S組相比,OJ組HO-1m RNA表達(dá)增加(P0.05);與OJ組相比,D組HO-1mRNA表達(dá)進(jìn)一步增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。D+Z組與D組比較HO-1mRNA表達(dá)減少,差異有統(tǒng)計(jì)學(xué)意義(P0.05),但與OJ組比較差異無(wú)意義(P0.05)。結(jié)論:1.右美托咪定可通過(guò)抑制氧化應(yīng)激反應(yīng)減少梗阻性黃疸大鼠肝臟損傷。2.右美托咪定通過(guò)上調(diào)HO-1mRNA及其蛋白表達(dá)對(duì)梗阻性黃疸導(dǎo)致的肝損傷起保護(hù)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.

【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R614
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本文編號(hào)：1842603