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二乙基亚硝胺诱发大鼠肝癌过程中肝脏胶原的动态观察
周陈杰,宫绪萌,蔡理,汪艳,高毅*
0
(南方医科大学珠江医院肝胆二科, 广州 510282
*通信作者)
摘要:
目的 动态观察大鼠肝脏癌变过程中胶原纤维的变化特点,为肝癌发病机制的研究提供参考。 方法 将50只雄性Wistar大鼠(体质量100~120 g)随机分为模型组和对照组,分别腹腔注射二乙基亚硝胺(DEN)50 mg/kg(加生理盐水至0.1 mL)或生理盐水(0.1 mL),均每周2次,连续4周后改为每周1次,至14周停止。处死大鼠,取肝脏病变组织及周边正常组织,通过H-E染色、Masson染色、网状纤维染色观察其形态学改变;采用荧光定量PCR技术检测大鼠肝组织Ⅰ型及Ⅲ型胶原mRNA的动态表达;采用明胶酶谱法检测大鼠肝组织中基质金属蛋白酶(MMP)含量变化。 结果 DEN诱癌开始5周后大鼠肝组织肝硬化形成,14周后诱导出肝癌。相应地,肝组织内胶原沉积持续增加,癌灶内胶原含量则明显少于癌周组织,并呈进行性减少;MMP-2、MMP-9在癌周组织和癌组织中的变化趋势则与胶原含量变化相反。 结论 DEN诱导的大鼠肝脏癌变过程中,胶原纤维在肝组织中沉积增加,在肝癌组织中则减少,随着肝癌的进展,癌组织内胶原进一步减少,提示肝硬化组织癌变过程中胶原纤维可能被降解。
关键词:  肝肿瘤  肝硬化  二乙基亚硝胺  胶原纤维
DOI:10.3724/SP.J.1008.2014.00626
投稿时间:2014-01-23修订日期:2014-05-17
基金项目:
Sequential study of liver collagen fibers in diethylnitrosamine-induced hepatocarcinogenesis in rats
ZHOU Chen-jie,GONG Xu-meng,CAI Li,WANG Yan,GAO Yi*
(Department of Hepatobiliary Surgery, Zhujiang Hospital, Nanfang Medical University, Guangzhou 510282, Guangdong, China
*Corresponding authors.)
Abstract:
Objective To observe the sequential changes of collagen fibers in rats with diethylnitrosamine (DEN)-induced hepatocellular carcinoma (HCC), so as to provide a reference for pathogenesis research of HCC. Methods Fifty male Wistar rats, weighing 100-120 g, were randomly divided into normal group and HCC model group. The model group was intraperitoneally administered with 50 mg/kg DEN (0.1 mL), twice a week for 4 weeks, then once a week for another 10 weeks. The control group was given normal saline (0.1 mL) in the same manner. Finally the rats were sacrificed; the normal and diseased liver tissues were observed by H-E, Masson and argyrophilic fiber staining. The expression of collagen type Ⅰ and type Ⅲ mRNA was detected by fluorogenic quantitative PCR; the expression of matrix metalloproteinase (MMP) was examined by gelatinases spectrometry. Results Cirrhosis was found in rats 5 weeks after treatment with DEN and HCC was induced 14 weeks after DEN treatment; collagen deposition in liver tissues increased in a progressive manner, and the collagen contents in the HCC tissues was greatly less than that in the adjacent tissues, showing a decreasing trend. The contents of MMP-2 and MMP-9 in the HCC and adjacent tissues had opposite changes compared with collagen. Conclusion Collagen deposition in cirrhosis liver tissue is increased during the process of DEN-induced HCC, but it is decreased in HCC tissues in a progressive manner, which indicates that collagen might be degraded during the progression from cirrhosis to HCC.
Key words:  liver neoplasms  cirrhosis  diethylnitrosamine  collagen fiber