【摘要】：脂聯(lián)素是近來(lái)發(fā)現(xiàn)的一種脂肪組織特異性產(chǎn)生的細(xì)胞因子,它除了調(diào)節(jié)糖脂代謝、改善胰島素抵抗外,還能通過(guò)抑制腫瘤壞死因子α(TNF-α)的產(chǎn)生與作用來(lái)調(diào)節(jié)機(jī)體的炎癥反應(yīng)。病毒性肝炎的主要致病機(jī)理就是活化的T細(xì)胞產(chǎn)生以TNF-α為主的細(xì)胞因子介導(dǎo)對(duì)肝細(xì)胞的免疫性損傷。因此,本文采用Con A誘導(dǎo)性肝損傷小鼠模型進(jìn)行脂聯(lián)素干預(yù)實(shí)驗(yàn),以探討脂聯(lián)素是否對(duì)肝臟的免疫性損傷具有保護(hù)作用。研究的主要內(nèi)容包括:①?gòu)闹窘M織克隆小鼠脂聯(lián)素基因,并在大腸桿菌內(nèi)表達(dá);②構(gòu)建小鼠脂聯(lián)素真核表達(dá)載體pAA-neo-mAd,并在COS-7細(xì)胞中驗(yàn)證其表達(dá);③將表達(dá)載體pAA-neo-mA與pCI-neo-mAd導(dǎo)入小鼠體內(nèi),驗(yàn)證它們?cè)诟蝺?nèi)的表達(dá)及表達(dá)效率;④在將載體pAA-neo-mAd導(dǎo)入小鼠體內(nèi),并獲得重組脂聯(lián)素在肝臟內(nèi)的高效穩(wěn)定表達(dá)的基礎(chǔ)上,進(jìn)行Con A致小鼠免疫性肝損傷的脂聯(lián)素干預(yù)實(shí)驗(yàn),通過(guò)肝臟組織學(xué)檢查、血清ALT、TNF-α濃度測(cè)定等比較表達(dá)載體組與空載體組(陰性對(duì)照)肝損傷程度的不同,并分析脂聯(lián)素、TNF-α及ALT之間的相關(guān)性。 研究結(jié)果發(fā)現(xiàn),表達(dá)載體組小鼠的肝臟損傷明顯輕于空載體組(兩組ALT比較,P0.0001),其血清TNF-α濃度也低于空載體組(P0.0001)。初步表明脂聯(lián)素對(duì)Con A導(dǎo)致的肝臟免疫性損傷具有保護(hù)作用,其作用機(jī)制可能是通過(guò)抑制TNF-α的產(chǎn)生。 本研究將流體動(dòng)力法(Hydrodynamics-based procedure)應(yīng)用于小鼠脂聯(lián)素的體內(nèi)表達(dá),成功地將脂聯(lián)素真核表達(dá)載體導(dǎo)入了小鼠肝臟,獲得了重組脂聯(lián)素在肝內(nèi)的高效穩(wěn)定表達(dá)。并通過(guò)脂聯(lián)素對(duì)Con A致肝臟免疫性損傷的干預(yù)實(shí)驗(yàn)初步表明,脂聯(lián)素對(duì)Con A導(dǎo)致的T細(xì)胞介導(dǎo)的肝臟免疫性損傷具有保護(hù)作用。這一發(fā)現(xiàn)為脂聯(lián)素在病毒性肝炎臨床防治方面的研究提供了重要的理論依據(jù)。
[Abstract]:Adiponectin is a recently discovered cytokine specifically produced by adipose tissue. In addition to regulating glucose and lipid metabolism, adiponectin improves insulin resistance. It can also regulate the inflammatory response by inhibiting the production and action of tumor necrosis factor 偽 (TNF- 偽). The main pathogenetic mechanism of viral hepatitis is the immune damage induced by activated T cells by cytokines mediated by TNF- 偽. In order to investigate whether adiponectin has protective effect on immune injury of liver, adiponectin was used to induce liver injury in Con A mice. The main contents of this study were as follows: (1) the adiponectin gene was cloned from adipose tissue and expressed in E. coli; (2) the eukaryotic expression vector of mouse adiponectin (pAA-neo-mAd,) was constructed and its expression was verified in COS-7 cells. (3) the expression vectors pAA-neo-mA and pCI-neo-mAd were introduced into mice to verify their expression and expression efficiency in liver. 4 on the basis of introducing the vector pAA-neo-mAd into mice and obtaining the high and stable expression of recombinant adiponectin in the liver, the adiponectin intervention experiment of immune liver injury induced by Con A in mice was carried out. The liver histological examination and serum ALT, were carried out. The degree of liver injury was compared between the expression vector group and the empty vector group (negative control group), and the correlation among adiponectin, TNF- 偽 and ALT was analyzed. The results showed that the liver injury in the expression vector group was significantly lighter than that in the empty vector group (P 0.0001), and the serum TNF- 偽 concentration was lower in the expression vector group than in the empty vector group (P 0.0001). It is suggested that adiponectin has protective effect on hepatic immune injury induced by Con A, and its mechanism may be by inhibiting the production of TNF- 偽. In this study, the hydrodynamic method (Hydrodynamics-based procedure) was applied to the expression of adiponectin in mice. The eukaryotic expression vector of adiponectin was successfully introduced into mouse liver, and the high and stable expression of recombinant adiponectin in the liver was obtained. The intervention of adiponectin on Con A induced liver immune injury showed that adiponectin had protective effect on T cell mediated hepatic immune injury induced by Con A. This discovery provides an important theoretical basis for the study of adiponectin in clinical prevention and treatment of viral hepatitis.
【學(xué)位授予單位】：中國(guó)人民解放軍軍事醫(yī)學(xué)科學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2005
【分類號(hào)】：R392

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 ;Insulin expression in livers of diabetic mice mediated by hydrodynamics-based administration[J];World Journal of Gastroenterology;2004年04期

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本文編號(hào)：2402929