慢性乙型肝炎肝脏炎症和纤维化中CaM、Bad蛋白变化影响的研究

发布时间：2018-06-10 04:14

本文选题：凋亡因子 + 炎症　；参考：《郑州大学》2013年硕士论文

【摘要】：背景凋亡(apoptosis)是个别细胞程序性细胞死亡的表现形式,是由体内外因素出发细胞内预存的死亡程序而导致的细胞主动性死亡方式。凋亡在生物胚胎发生发育、成熟细胞新旧交替、激素依赖性生理退化、萎缩和老化以及化学诱导的细胞死亡都发挥着不可替代的重要作用。乙型肝炎病毒(HBV)慢性感染是我国重大的公共卫生问题,尽管我国长期以来在肝炎防治中取得显著成效,但HBV慢性感染仍是危害最严重的传染病之一,肝细胞凋亡在慢性乙型肝炎患者病情进展中起着举足轻重的地位,碎片状坏死是慢性乙型肝炎早期特征性病理改变,毛玻璃样变是肝炎、肝纤维化及肝硬化的必然进程,目前已证实,该病理改变实质即为凋亡。经典的凋亡途径包括两种,即内源性途径(intrinsic pathway,又称线粒体凋亡途径)和外源性途径(extrinsic pathway,又称死亡受体凋亡途径)。近年来研究证明,除线粒体以外,内质网(endoplasmic reticulum, ER)等细胞器也成为细胞凋亡信号整合的主要位点。研究发现,ER可能是细胞内诱导凋亡的一个场所,因此,内质网应激(endoplasmic reticulum stress, ERS)反应性凋亡途径作为一种新的凋亡途径,成为凋亡研究的热点之一。慢性乙型肝炎病理分级(grading, G)与分期(staging, S)是临床上评估肝脏炎症活动程度和纤维增生程度的指标,其中乙型肝炎病毒是诱发肝脏炎症和纤维化的始动因素,肝细胞转归主要通过细胞坏死和凋亡,其中不同炎症程度诱发细胞凋亡是肝细胞受到影响的重要因素,然而在不同程度下肝脏炎症和纤维化之间的变化关系对细胞凋亡影响如何,目前见到报道较少,本文着重采用免疫组织化学SP法观察内质网凋亡途径中两种关键的细胞凋亡因子：钙调蛋白(calmodulin, CaM)和Bad蛋白在慢性乙型肝炎患者不同病理分级和分期的表达情况,通过肝脏炎症分级和纤维化分期影响两种凋亡因子,以期更深一步地了解肝脏病理进展中肝细胞凋亡的信号转导变化,分析内质网凋亡途径在慢性乙型肝炎中的地位和意义,为慢性乙型肝炎的治疗提供理论基础。目的观察比较CaM和Bad在慢性乙型肝炎(Chronic Hepatitis B, CHB)不同炎症分级和纤维化分期中的表达情况,探讨两种因子在不同肝脏炎症活动和纤维增生程度中的变化规律。方法病例收集：收集2010年6月至2012年12月在郑州大学第一附属医院感染科确诊为CHB并同期进行肝组织病理学检查的病例,排除合并其他疾病如其他病毒性肝炎、肝硬化失代偿期、酒精性肝病、脂肪肝、自身免疫性肝炎、原发性胆汁性肝硬化、肝癌；或合并严重的心、脑、肺、肾等脏器实质性病变。实验方法：根据病理科医师诊断的不同分级分期,取肝脏活组织标本制作蜡块,进行苏木素-伊红HE染色,采用免疫组织化学SP法,检测CaM和Bad蛋白在肝脏的表达水平,采集图像后采用软件分析系统计算平均灰度值。统计学分析：采用SPSS17.0与EXCEL2007统计软件进行数据分析,计量资料以均数±标准差(x±s)表示。采用秩和检验比较不同病理分级分期凋亡因子表达的差异性,采用Spearman等级相关分析法判断肝细胞凋亡因子与肝脏病理学分级之间的相关性。结果 1.炎症分级G0、G1、G2、G3、G4间的CaM表达水平分别为86.570±2.00,115.827±9.344,136.389±8.694,147.111±10.266,170.904±5.060,秩和检验提示5组炎症分级之间CaM表达水平的差别有统计学意义(P0.05)。 2.纤维化分期S0、S1、S2、S3、S4间的CaM表达水平分别为82.939±6.300,118.318±9.119,137.439±±4.963,145.81±±7.082,169.919±10.258,秩和检验提示5组纤维化分期之间CaM表达水平的差别有统计学意义(P0.05)。 3.炎症分级G0、G1、G2、G3、G4间的Bad表达水平分别为72.849±±2.287,105.780±±2.672,121.206±±5.423,137.649±±5.953,166.435±±3.900,秩和检验提示5组炎症分级之间Bad表达水平的差别有统计学意义(P0.05)。 4.纤维化分期S0、S1、S2、S3、S4间的Bad表达水平分别为72.115±±3.325,105.454±4.815,121.332±±5.069,135.812±6.148,168.071±3.651,秩和检验提示5组纤维化分期之间Bad表达水平的差别有统计学意义(P0.05)。 5.CaM与炎症分级、纤维化分期相关,相关系数分别为0.332,0.361。 6.Bad与炎症分级、纤维化分期相关,相关系数分别为0.429,0.432。 7.炎症分级、纤维化分期是CaM与Bad肝细胞表达水平的独立危险因素。结论不同肝脏病理分级CaM的表达水平不同,CaM与肝脏的炎症和纤维化程度呈正相关。同时不同肝脏病理分级Bad的表达水平不同,Bad与肝脏的炎症和纤维化程度也呈正相关,表明在慢性乙型肝炎中,炎症或者纤维化程度级别的递增,都独立对内质网凋亡途径具有明显促进作用,因此内质网介导的肝细胞凋亡在肝脏病理进展中可能起重要作用,为临床治疗提供理论依据。
[Abstract]:Background

Apoptosis is the expression of cell death in individual cells , and is a cell - active death method caused by the internal and external factors . Apoptosis plays an important role in the development of biological embryos , new and old cells of mature cells , hormone - dependent physiological degeneration , atrophy and aging , and chemical - induced cell death .

Hepatitis B virus ( HBV ) chronic infection is a major public health problem in China , but chronic hepatitis B infection is one of the most serious infectious diseases . In recent years , it has been proved that , in addition to mitochondria , the organelle , such as endoplasmic reticulum ( ER ) , is the main site of apoptosis signal integration . It is found that ER may be a place to induce apoptosis in cells . Therefore , the apoptotic pathway of endoplasmic reticulum stress ( ERS ) is one of the hot spots of apoptosis .

Chronic hepatitis B pathological grading ( grading , G ) and staging ( S ) are the indexes of evaluating the degree of hepatic inflammation and fibrosis .

Purpose

To investigate the expression of CaM and Bad in different stages of inflammation and fibrosis of chronic hepatitis B ( B ) , and to explore the changes of the two factors in different liver inflammatory activities and degree of fibrous hyperplasia .

method

Case collection : collection of cases of diagnosis of liver histopathology in the First Affiliated Hospital of Zhengzhou University from June 2010 to December 2012 , excluding other diseases such as other viral hepatitis , decompensated liver cirrhosis , alcoholic liver disease , fatty liver , autoimmune hepatitis , primary biliary cirrhosis , and liver cancer ;
or combined with severe heart , brain , lung , kidney and other visceral organs .

Methods : The liver biopsy specimens were used to make wax block according to different stages of the diagnosis of the medical practitioners . The staining was carried out by using the immunohistochemical SP method . The expression level of CaM and Bad protein in the liver was detected , and the average gray value was calculated by the software analysis system after the image was acquired .

Statistical analysis : The data were analyzed by SPSS 17.0 and EXCEL2007 statistical software . The data were expressed by mean 卤 standard deviation ( x 卤 s ) . The correlation between hepatocyte apoptosis factor and liver pathological grade was determined by means of rank sum test .

Results

1 . The levels of CaM expression in G0 , G1 , G2 , G3 , G4 were 86.570 卤 2.00 , 115.827 卤 9.344 , 136.389 卤 8.694 , 147.111 卤 10.266 , 170.904 卤 5.060 respectively .

2 . The levels of CaM expression between stages S0 , S1 , S2 , S3 and S4 were 82.939 卤 6.300 , 118.318 卤 9.119 , 137.439 卤 4.963 , 145.81 卤 7.082 , 169.919 卤 10.258 , respectively .

3 . Bad expression levels were 72.849 卤 2.287 , 105.780 卤 2.672 , 121.206 卤 5.423 , 137.649 卤 5.953 , 166.435 卤 3.900 , 147.649 卤 5.953 , 166.435 卤 3.900 , P < 0.05 ) .

4 . Bad expression levels between stages S0 , S1 , S2 , S3 and S4 were 72.115 卤 3.325 , 105.454 卤 4.815 , 121.332 卤 5.69 , 135.812 卤 6.148 , 168.071 卤 3.651 , and the rank sum test suggested that there was significant difference in Bad expression level between 5 groups of fibrosis stages ( P0.05 ) .

5 . CaM was correlated with inflammation grade and fibrosis stage . The correlation coefficients were 0.332 and 0.361 , respectively .

6 . Bad was correlated with inflammation grade and fibrosis stage . The correlation coefficients were 0.429 and 0.432 , respectively .

7 . Inflammatory stage , stage of fibrosis is independent risk factor of CaM and Bad hepatocyte expression level .

Conclusion

The expression levels of CaM in different liver and pathological grades are different , CaM and liver inflammation and fibrosis degree are positively correlated . At the same time , the expression levels of Bad and liver inflammation and fibrosis are positively correlated , indicating that in the chronic hepatitis B , the level of inflammation or fibrosis promotes independently , and therefore , the endoplasmic reticulum - mediated apoptosis may play an important role in the pathological progress of the liver , thus providing a theoretical basis for clinical treatment .
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R512.62;R575.2

【参考文献】