Objective To examine the occurrence and possible mechanism of hepatocyte apoptosis in acute biliary tract infection. Methods After adaptive feeding for seven days, 30 male rats were randomly divided into five groups (n = 6 each) , i. e., one control group injected with 0. 2 ml of saline through the common bile duct and four experimental groups injected with an equal volume of Escherichia coli cell suspension to build an acute biliary tract infection model. Animals of the control group were killed 6 h after injection of saline, and those of the four experimental groups were killed respectively at the time points of 6, 12, 24, and 48 h after injection of E. coli. Liver tissue specimens were taken from all the rats to prepare tissue sections. Pathological features were examined by hematoxylin- eosin staining and light microscopy. Hepatocyte apoptosis was examined by cell counting after TUNEL staining and calculation of the apoptotic index. The expression of apoptosis- related proteins (Bcl- 2 and Bax) was measured by immunohistochemistry. Data comparison between groups was performed by analysis of variance, and pairwise comparisons were further conducted using the LSD- t test and Dunnett's T3 test. Results Tissue specimens of the control groups had no significant pathological changes, while those of the experimental groups showed hepatocyte apoptosis.Compared with the control group, the four experimental groups had significantly greater values of apoptotic index (P < 0. 05) . Additionally, the values of apoptotic index in the 24 and 48 h experimental groups significantly increased over time compared with that of the 12 h experimental group (P < 0. 05) . Bax protein expression level significantly differed between the control group and the experimental groups ( P<0. showing="" a="" slight="" increasing="" trend="" over="" time="" in="" the="" latter="" four="" groups="" p="">0. 05) . Bcl- 2 protein expression level had no significant differences between the control group and the 6 or 12 h experimental groups (P > 0. 05) . Over time, however, significant differences were observed in Bcl- 2 expression level between the 12 h experimental group and the 24 or 48 h experimental group (P < 0. 05) . Conclusion Hepatocyte apoptosis occurs within a short time after acute biliary tract infection. It persists in the whole process of the infection, with increased expression of Bax (inducer of apoptosis) and decreased expression of Bcl- 2 (inhibitor of apoptosis) . The changes in the expression of apoptosis- related genes coincide with hepatocyte apoptosis, suggesting that hepatocyte apoptosis may be associated with Bax and Bcl-2 genes.
[1]KERR JFR, WYLLIE AH, CURRIE AR.Apoptosis:a basic biological phenomenon with wide ranging implications in tissue kinetics[J].Br J Cancer, 1972, 26 (4) :239-257.
|
[2]LAHORTE CM, VANDERHEYDEN JL, STEINMETZ N, et al.Apoptosis-detecting radioligands:current state of the art and future perspectives[J].Eur J Nucl Med Mol Imaging, 2004, 31 (6) :887-919.
|
[3]XIA XM, HE K, FENG CH, et al.Cholangitis and hepatocyte apoptosis and expression of apoptosis gene[J].China J Modern Med, 2006, 16 (2) :233-236, 240. (in Chinese) 夏先明, 贺凯, 冯春红, 等.胆道感染与肝细胞凋亡及凋亡相关基因的表达[J].中国现代医学杂志, 2006, 16 (2) :233-236, 240.
|
[4]LALISANG TJ, SJAMSUHIDAJAT R, SIREGAR NC, et al.Profile of hepatocyte apoptosis and bile lakes before and after bile duct decompression in severe obstructive jaundice patients[J].Hepatobiliary Pancreat Dis Int, 2010, 9 (5) :520-523.
|
[5]FICKERT P, TRAUNER M, FUCHSBICHLER A, et al.Oncosis represents the main type of cell death in mouse models of cholestasis[J].J Hepatol, 2005, 42 (3) :378-385.
|
[6]MALHI H, GORES G J, LEMASTERS JJ.Apoptosis and necrosis in the liver:A tale of two deaths[J].Hepatology, 2006, 43 (2Suppl 1) :s31-s44.
|
[7]LASTER SM, GOOD JG, GOODING LR, et al.Tumor necrosis factor can induce both apoptic and necrotic forms of cell lysis[J].J Immunol, 1988, 141 (8) :2629-2634.
|
[8]WANG JX, WANG GJ, CAI X.The effects of oxymatrine on apoptosis induced by TNF-αin human liver cell line L02[J].J Clin Hepatol, 2011, 17 (4) :210-221. (in Chinese) 王俊学, 王国俊, 蔡雄.氧化苦参碱对肿瘤坏死因子诱导的人L02肝细胞凋亡的影响[J].临床肝胆病杂志, 2011, 17 (4) :210-221.
|
[9]LEIST M, GANTER F, BOHLINGER I, et al.Murine hepatocyte apoptosis induced in vitro and in vivo by TNF-alpha requires transcriptional arrest[J].J Immunol, 1994, 153 (4) :1778-788.
|
[10]YAO YZ, BAO ZJ.Diagnostic value of serum IL-6, IL-8 and TNF-αin monitoring inflammation after operation of biliary tract infection[J].J Med Res, 2012, 41 (4) :104-106. (in Chinese) 姚燕珍, 鲍舟君.血清IL-6、IL-8和TNF-α在胆道术后感染监测中的价值[J].医学研究杂志, 2012, 41 (4) :104-106.
|
[11]LEON CG, TORY R, JIA J, et al.Discovery and development of tolllike receptor 4 (TLR4) antagonists:a new paradigm for treating sepsis and other diseases[J].Pharm Res, 2008, 25 (8) :1751-1761.
|
[12]YI JM, GUAN YS.Process and mechanism of inflammatory response following biliary obstruction[J].Chin J Gen Surg, 2012, 21 (5) :607-610. (in Chinese) 易杰明, 关养时.胆道梗阻后炎症反应发生的过程及机制[J].中国普通外科杂志, 2012, 21 (5) :607-610.
|
[13]CHEN G, ZHOU SQ.Difference in the expression of iNOS in hepatocytes of rat with biliary tract infection and with abdominal infection[J].Chin J Exp Surg, 2003, 20 (12) :1083-1084. (in Chinese) 陈刚, 邹声泉.胆道感染与腹腔感染大鼠肝细胞诱生型一氧化氮合酶表达差异的实验研究[J].中华实验外科杂志, 2003, 20 (12) :1083-1084.
|
[14]ZHAO Y, LI L, YIN Q.Effects of iNOS-derived NO on Bcl-2and Bax protein expression following focal cerebral ischemia and reperfusion in rats[J].Chin J Neuroanatomy, 2005, 21 (5) :539-542. (in Chinese) 赵昱, 李莉, 尹青.大鼠局灶性脑缺血再灌注中iNOS源性NO对Bcl-2, Bax表达的影响[J].神经解剖学杂志, 2005, 21 (5) :539-542.
|
[15]MATSUDA T, SAITO H, FUKATSU K, et al.Cytokine-modulated inhibition of neutrophil apoptosis at local site augments exudative neutrophil functions and reflects inflammatory response after surgery[J].Surgery, 2001, 129 (1) :76-85.
|
[16] YOULE RJ, STRASSER A.The Bcl-2 protein family:opposing activities that mediate cell death[J].Nat Rev Mol Cell Biol, 2008, 9 (1) :47-59.
|
[17]WANG JT, GONG SS.Effects of siRNA specific to the protein kinase CK2αon apoptosis of laryngeal carcinoma cells[J].Chin Med J (Engl) , 2012, 125 (9) :1581-1585.
|
[18]KOBAYASHI T, SAWA H, MORIKAWA J, et al.Bax induction activates apoptotic cascade via mitochondrial cytochrome C release and bax overexpression enhances apoptosis induced by chemotherapeutic agents in dld-1 colon cancer cells[J].Jpn J Cancer Res, 2000, 91 (12) :1264-1268.
|
[19]SPRICK MR, WALCZAK H.The interplay between the Bcl-2family and death receptor-mediated apoptosis[J].Biochim Biophys Acta, 2004, 1644 (2-3) :125-132.
|