慢性炎症损伤在二乙基亚硝胺诱导大鼠肝细胞癌发生中的作用  

The role of chronic inflammatory injury in the occurrence of hepatocellular carcinoma induced by diethylnitrosamine in rats

作　　者：张昊[1] 李文[1] 陈署贤[1] 潘楚芝[1] 王楚斯 潘卫东[1] Zhang Hao Li Wen Chen Shuxian Pan Chuzhi Wang Chusi Pan Weidong.(Department of Hepatobiliary Surgery, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China)

机构地区：[1]中山大学附属第三医院肝胆外科,广州510630

出　　处：《中华肝脏外科手术学电子杂志》2017年第4期332-336,共5页Chinese Journal of Hepatic Surgery(Electronic Edition)

基　　金：国家自然科学基金(81172783)

摘　　要：目的探讨慢性炎症损伤在二乙基亚硝胺(DEN)诱导大鼠肝细胞癌(肝癌)发生中的作用。方法健康雄性Wistar大鼠40只,按随机数字表法随机分为亚硝胺组(32只)及对照组(8只)。亚硝胺组腹腔注射DEN 60 mg/kg,1次/周,连续8周;对照组不给药。亚硝胺组于第4、8、12、16周随机取8只大鼠的肝组织行病理学切片,预留门静脉血,第16周处死所有大鼠。HE染色观察肝组织炎症损伤情况,免疫组化法检测肝组织DNA修复蛋白γH2AX的表达,ELISA检测血清炎症因子IL-6、VEGF、TNF-α的表达水平。3组血清炎症因子比较采用单因素方差分析和LSD-t检验。两组γH2AX阳性率比较采用单样本t检验或独立样本t检验。结果亚硝胺组大鼠第4、8、12、16周肝组织病理炎症损伤逐渐加重,并最终发展为肝癌;对照组未见异常。亚硝胺组大鼠第4、8、12、16周肝组织γH2AX阳性率分别为(0.3±0.1)%、(3.1±0.6)%、(7.8±2.8)%、(11.4±2.3)%,明显高于对照组的0(Z=2.34,2.84,4.11,5.31;P<0.05)。亚硝胺组第4、8、12、16周血清IL-6水平分别为(104±7)、(117±10)、(168±16)、(181±11)ng/L,明显高于对照组的(83±53)ng/L(LSD-t=5.19,8.41,21.03,24.25;P<0.05)。亚硝胺组第4、8、12、16周血清VEGF水平分别为(257±16)、(302±21)、(387±36)、(439±26)ng/L,明显高于对照组的(164±15)ng/L(LSD-t=10.17,15.04,24.39,29.18;P<0.05)。亚硝胺组第4、8、12、16周血清TNF-α水平分别为(108±5)、(125±11)、(156±17)、(181±14)ng/L,明显高于对照组的(79±6)ng/L(LSD-t=7.30,11.59,19.39,25.70;P<0.05)。结论肝脏慢性炎症损伤在DEN诱导大鼠肝癌的发生中起重要作用,此过程伴随着高水平的炎症因子和持续的DNA损伤。Objective To investigate the role of chronic inflammatory injury in the occurrence of hepatocellular carcinoma （HCC） induced by diethylnitrosamine （DEN） in rats. Methods Forty healthy male Wistar rats were randomly divided into the DEN group （n=32） and control group （n=8） according to the random number table method. DEN （60 mg/kg） was administered in the DEN group via intraperitoneal injection once weekly for consecutive 8 weeks, and no drug was administered in the control group. In the DEN group, the liver tissues were obtained from 8 rats randomly for pathological examination at 4, 8, 12 and 16 week, and the portal venous blood was preserved. All rats were executed at 16 week. The inflammatory injury of liver tissues was observed by HE staining. The expression level of DNA repair protein γH2AX was detected by immunohistochemical staining. The expression levels of IL-6, VEGF and TNF-α were detected by ELISA. Serum inflammatory factors were compared among three groups using one-way analysis of variance and LSD-t test. The positive rate of γH2AX was compared between two groups using one-sample t test or independent-sample t test. Results In the DEN group, the inflammatory injury of liver tissues was gradually aggravated at 4, 8, 12 and 16 week, and eventually progressed into HCC. No pathological abnormality was observed in the control group. The positive rate of γH2AX in the liver tissues at 4, 8, 12 and 16 week was respectively （0.3±0.1）%, （3.1±0.6）%, （7.8±2.8）% and （11.4±2.3）% in the DEN group, significantly higher than 0 in the control group （Z=2.34, 2.84, 4.11, 5.31; P〈0.05）. The level of serum IL-6 at 4, 8, 12 and 16 week was respectively （104±7）, （117±10）, （168±16） and （181±11） ng/L in the DEN group, significantly higher than （83±53） ng/L in the control group （LSD-t=- 5.19, 8.41, 21.03, 24.25; P〈0.05）. The level of serum VEGF at 4, 8, 12 and 16 week was respectively （257±16）, （302±21）, （387±36） and （439±26） n

关 键 词：癌 肝细胞 慢性 肝损伤 二乙基亚硝胺 γH2AX 炎症 

分 类 号：R735.7[医药卫生—肿瘤]