本文選題：凋亡因子 + 炎癥��； 參考：《鄭州大學(xué)》2013年碩士論文

【摘要】：背景 凋亡(apoptosis)是個(gè)別細(xì)胞程序性細(xì)胞死亡的表現(xiàn)形式,是由體內(nèi)外因素出發(fā)細(xì)胞內(nèi)預(yù)存的死亡程序而導(dǎo)致的細(xì)胞主動(dòng)性死亡方式。凋亡在生物胚胎發(fā)生發(fā)育、成熟細(xì)胞新舊交替、激素依賴性生理退化、萎縮和老化以及化學(xué)誘導(dǎo)的細(xì)胞死亡都發(fā)揮著不可替代的重要作用。 乙型肝炎病毒(HBV)慢性感染是我國(guó)重大的公共衛(wèi)生問題,盡管我國(guó)長(zhǎng)期以來(lái)在肝炎防治中取得顯著成效,但HBV慢性感染仍是危害最嚴(yán)重的傳染病之一,肝細(xì)胞凋亡在慢性乙型肝炎患者病情進(jìn)展中起著舉足輕重的地位,碎片狀壞死是慢性乙型肝炎早期特征性病理改變,毛玻璃樣變是肝炎、肝纖維化及肝硬化的必然進(jìn)程,目前已證實(shí),該病理改變實(shí)質(zhì)即為凋亡。經(jīng)典的凋亡途徑包括兩種,即內(nèi)源性途徑(intrinsic pathway,又稱線粒體凋亡途徑)和外源性途徑(extrinsic pathway,又稱死亡受體凋亡途徑)。近年來(lái)研究證明,除線粒體以外,內(nèi)質(zhì)網(wǎng)(endoplasmic reticulum, ER)等細(xì)胞器也成為細(xì)胞凋亡信號(hào)整合的主要位點(diǎn)。研究發(fā)現(xiàn),ER可能是細(xì)胞內(nèi)誘導(dǎo)凋亡的一個(gè)場(chǎng)所,因此,內(nèi)質(zhì)網(wǎng)應(yīng)激(endoplasmic reticulum stress, ERS)反應(yīng)性凋亡途徑作為一種新的凋亡途徑,成為凋亡研究的熱點(diǎn)之一。 慢性乙型肝炎病理分級(jí)(grading, G)與分期(staging, S)是臨床上評(píng)估肝臟炎癥活動(dòng)程度和纖維增生程度的指標(biāo),其中乙型肝炎病毒是誘發(fā)肝臟炎癥和纖維化的始動(dòng)因素,肝細(xì)胞轉(zhuǎn)歸主要通過(guò)細(xì)胞壞死和凋亡,其中不同炎癥程度誘發(fā)細(xì)胞凋亡是肝細(xì)胞受到影響的重要因素,然而在不同程度下肝臟炎癥和纖維化之間的變化關(guān)系對(duì)細(xì)胞凋亡影響如何,目前見到報(bào)道較少,本文著重采用免疫組織化學(xué)SP法觀察內(nèi)質(zhì)網(wǎng)凋亡途徑中兩種關(guān)鍵的細(xì)胞凋亡因子：鈣調(diào)蛋白(calmodulin, CaM)和Bad蛋白在慢性乙型肝炎患者不同病理分級(jí)和分期的表達(dá)情況,通過(guò)肝臟炎癥分級(jí)和纖維化分期影響兩種凋亡因子,以期更深一步地了解肝臟病理進(jìn)展中肝細(xì)胞凋亡的信號(hào)轉(zhuǎn)導(dǎo)變化,分析內(nèi)質(zhì)網(wǎng)凋亡途徑在慢性乙型肝炎中的地位和意義,為慢性乙型肝炎的治療提供理論基礎(chǔ)。 目的 觀察比較CaM和Bad在慢性乙型肝炎(Chronic Hepatitis B, CHB)不同炎癥分級(jí)和纖維化分期中的表達(dá)情況,探討兩種因子在不同肝臟炎癥活動(dòng)和纖維增生程度中的變化規(guī)律。 方法 病例收集：收集2010年6月至2012年12月在鄭州大學(xué)第一附屬醫(yī)院感染科確診為CHB并同期進(jìn)行肝組織病理學(xué)檢查的病例,排除合并其他疾病如其他病毒性肝炎、肝硬化失代償期、酒精性肝病、脂肪肝、自身免疫性肝炎、原發(fā)性膽汁性肝硬化、肝癌；或合并嚴(yán)重的心、腦、肺、腎等臟器實(shí)質(zhì)性病變。 實(shí)驗(yàn)方法：根據(jù)病理科醫(yī)師診斷的不同分級(jí)分期,取肝臟活組織標(biāo)本制作蠟塊,進(jìn)行蘇木素-伊紅HE染色,采用免疫組織化學(xué)SP法,檢測(cè)CaM和Bad蛋白在肝臟的表達(dá)水平,采集圖像后采用軟件分析系統(tǒng)計(jì)算平均灰度值。 統(tǒng)計(jì)學(xué)分析：采用SPSS17.0與EXCEL2007統(tǒng)計(jì)軟件進(jìn)行數(shù)據(jù)分析,計(jì)量資料以均數(shù)±標(biāo)準(zhǔn)差(x±s)表示。采用秩和檢驗(yàn)比較不同病理分級(jí)分期凋亡因子表達(dá)的差異性,采用Spearman等級(jí)相關(guān)分析法判斷肝細(xì)胞凋亡因子與肝臟病理學(xué)分級(jí)之間的相關(guān)性。 結(jié)果 1.炎癥分級(jí)G0、G1、G2、G3、G4間的CaM表達(dá)水平分別為86.570±2.00,115.827±9.344,136.389±8.694,147.111±10.266,170.904±5.060,秩和檢驗(yàn)提示5組炎癥分級(jí)之間CaM表達(dá)水平的差別有統(tǒng)計(jì)學(xué)意義(P0.05)。 2.纖維化分期S0、S1、S2、S3、S4間的CaM表達(dá)水平分別為82.939±6.300,118.318±9.119,137.439±±4.963,145.81±±7.082,169.919±10.258,秩和檢驗(yàn)提示5組纖維化分期之間CaM表達(dá)水平的差別有統(tǒng)計(jì)學(xué)意義(P0.05)。 3.炎癥分級(jí)G0、G1、G2、G3、G4間的Bad表達(dá)水平分別為72.849±±2.287,105.780±±2.672,121.206±±5.423,137.649±±5.953,166.435±±3.900,秩和檢驗(yàn)提示5組炎癥分級(jí)之間Bad表達(dá)水平的差別有統(tǒng)計(jì)學(xué)意義(P0.05)。 4.纖維化分期S0、S1、S2、S3、S4間的Bad表達(dá)水平分別為72.115±±3.325,105.454±4.815,121.332±±5.069,135.812±6.148,168.071±3.651,秩和檢驗(yàn)提示5組纖維化分期之間Bad表達(dá)水平的差別有統(tǒng)計(jì)學(xué)意義(P0.05)。 5.CaM與炎癥分級(jí)、纖維化分期相關(guān),相關(guān)系數(shù)分別為0.332,0.361。 6.Bad與炎癥分級(jí)、纖維化分期相關(guān),相關(guān)系數(shù)分別為0.429,0.432。 7.炎癥分級(jí)、纖維化分期是CaM與Bad肝細(xì)胞表達(dá)水平的獨(dú)立危險(xiǎn)因素。 結(jié)論 不同肝臟病理分級(jí)CaM的表達(dá)水平不同,CaM與肝臟的炎癥和纖維化程度呈正相關(guān)。同時(shí)不同肝臟病理分級(jí)Bad的表達(dá)水平不同,Bad與肝臟的炎癥和纖維化程度也呈正相關(guān),表明在慢性乙型肝炎中,炎癥或者纖維化程度級(jí)別的遞增,都獨(dú)立對(duì)內(nèi)質(zhì)網(wǎng)凋亡途徑具有明顯促進(jìn)作用,因此內(nèi)質(zhì)網(wǎng)介導(dǎo)的肝細(xì)胞凋亡在肝臟病理進(jìn)展中可能起重要作用,為臨床治療提供理論依據(jù)。
[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 鹵 standard deviation ( x 鹵 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 鹵 2.00 , 115.827 鹵 9.344 , 136.389 鹵 8.694 , 147.111 鹵 10.266 , 170.904 鹵 5.060 respectively .

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

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

4 . Bad expression levels between stages S0 , S1 , S2 , S3 and S4 were 72.115 鹵 3.325 , 105.454 鹵 4.815 , 121.332 鹵 5.69 , 135.812 鹵 6.148 , 168.071 鹵 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 .
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：R512.62;R575.2

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本文編號(hào)：2001935