多模式肝损伤后Hedgehog信号通路的表达分析
摘要：目的 初步探讨多种模式肝损伤后Hedgehog信号通路的表达及其变化,为其天然植物阻断剂和中药新药开发及中药治疗肝病提供依据。方法 雄性Fisher344大鼠140只适应性喂养1周后,随机分为药物组、手术组、干细胞增生组与正常对照组。按Gordon’s方案分次腹腔注射倒千里光碱或对照剂,手术组和干细胞增生组择期行部分肝切除术诱导急性肝损伤与干细胞增殖,药物组、正常对照组予假手术。实时荧光定量PCR、real-time PCR及免疫组化等方法检测原代肝细胞及组织切片中Hedgehog信号通路相关成分在不同时间点的表达。结果 多种因素所致肝损伤都可引起Hedgehog信号通路的异常激活,相关成分IHH、PTCH、Smo、Gli1、Gli2、Gli3 mRNA动态表达(SHH阴性),信号分子IHH及通路激活标志(PTCH、Gli1)3个指标在时间与处理组之间存在交互效应(均P<0.001)。免疫组化证实PTCH在蛋白水平表达,且不同损伤模式下表达持续的时间不同。结论 Hedgehog信号通路在各种肝损伤后异常激活,可能是参与调控肝损伤后反应的最重要的信号通路之一。
关键词：Hedgehog信号通路；肝损伤；肝再生；大鼠
DOI：10.3969/j.issn.1005-5304.2013.08.017
中图分类号：R363
文献标识码：A
文章编号：1005-5304(2013)08-0048-04Expression of Hedgehog Signaling Pathway after Different Liver Damage CAI Yi-dong1, WANG Li2, ZHENG Hao-xuan3, SUN Shu-mei1, WANG Yang1, LI Chun4 (1.Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing 100091, China；2.Shanxi University of TCM, Taiyuan 030024, China；3.Nanfang Hospital, Southern Medical University, Guangzhou 510515, China；4.China Academy of Chinese Medical Sciences, Beijing 100700, China)
Abstract：Objective To preliminarily explore the expresssion of the Hedgehog signaling pathway (HH) after different liver trauma, and provide basis for the blockaded effection of HH and the potentiality of new traditional Chinese medicine exploition. Methods After adaptive breeding for one week, a cohort of 140 male SPF Fisher 344 rats were randomly divided into Retrorsine injection (R) indicated the toxical of chemical damage of liver, partial hepatectomy (PH) indicated the acute liver trauma, retrorsine/partial hepatectomy (R/PH) for the hepatic stem cells regeneration, and normal control (N) groups. The administering of retrorsine or placebo, and PH or sham operation were performed as Gordon’s matheds. RT-PCR, real-time PCR and immunohistochemistry were performed to detect the expression of the constituents related to HH after liver damage. Results RT-PCR demonstrated that the mRNA expression of IHH, PTCH, SMO, Gli1, Gli2, Gli3 were all activated in primary liver cells after different kinds of liver damage. The results of real-time PCR analysis indicated an interaction effect existing between the treatment and the time in 3 Hedgehog-related constituents (signaling molecule IHH, indicators of the HH activation Ptc, Gli1) (all P<0.001). The protein expression of PTCH in liver tissue section was confirmed by immunohistochemistry, which differed in the duration in PH and R/PH groups. Conclusion The HH was abnormally activated during the different liver trauma. It’s maybe an important signaling pathway during the liver’s reaction after damage.
Key words：Hedgehog signalling pathway；liver trauma；liver regeneration；rats
在全球范围内,各种肝脏疾病尤其是终末期肝病(包括肝硬化和肝癌)的发病呈上升趋势[1-2]。对各种肝损伤后发病机理的研究,一直是肝病领域的热点。近年来,关于Hedgehog信号通路(hedgehog signaling pathway,HH)在肝脏疾病中的作用受到较多的关注[3-4]。HH由于存在天然植物阻断剂(藜芦类生物碱,环王巴明cyclopamine)[5-6],阐明其在肝损伤后反应过程中通讯作者：李淳,E-mail：lichunchris@hotmail.com的作用,寻找具有阻断作用的中药类似物,对开发中药新药具有重要的现实意义。, 百拇医药(蔡毅东 王丽 郑浩轩 孙淑美 王洋 李淳)
关键词：Hedgehog信号通路；肝损伤；肝再生；大鼠
DOI：10.3969/j.issn.1005-5304.2013.08.017
中图分类号：R363
文献标识码：A
文章编号：1005-5304(2013)08-0048-04Expression of Hedgehog Signaling Pathway after Different Liver Damage CAI Yi-dong1, WANG Li2, ZHENG Hao-xuan3, SUN Shu-mei1, WANG Yang1, LI Chun4 (1.Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing 100091, China；2.Shanxi University of TCM, Taiyuan 030024, China；3.Nanfang Hospital, Southern Medical University, Guangzhou 510515, China；4.China Academy of Chinese Medical Sciences, Beijing 100700, China)
Abstract：Objective To preliminarily explore the expresssion of the Hedgehog signaling pathway (HH) after different liver trauma, and provide basis for the blockaded effection of HH and the potentiality of new traditional Chinese medicine exploition. Methods After adaptive breeding for one week, a cohort of 140 male SPF Fisher 344 rats were randomly divided into Retrorsine injection (R) indicated the toxical of chemical damage of liver, partial hepatectomy (PH) indicated the acute liver trauma, retrorsine/partial hepatectomy (R/PH) for the hepatic stem cells regeneration, and normal control (N) groups. The administering of retrorsine or placebo, and PH or sham operation were performed as Gordon’s matheds. RT-PCR, real-time PCR and immunohistochemistry were performed to detect the expression of the constituents related to HH after liver damage. Results RT-PCR demonstrated that the mRNA expression of IHH, PTCH, SMO, Gli1, Gli2, Gli3 were all activated in primary liver cells after different kinds of liver damage. The results of real-time PCR analysis indicated an interaction effect existing between the treatment and the time in 3 Hedgehog-related constituents (signaling molecule IHH, indicators of the HH activation Ptc, Gli1) (all P<0.001). The protein expression of PTCH in liver tissue section was confirmed by immunohistochemistry, which differed in the duration in PH and R/PH groups. Conclusion The HH was abnormally activated during the different liver trauma. It’s maybe an important signaling pathway during the liver’s reaction after damage.
Key words：Hedgehog signalling pathway；liver trauma；liver regeneration；rats
在全球范围内,各种肝脏疾病尤其是终末期肝病(包括肝硬化和肝癌)的发病呈上升趋势[1-2]。对各种肝损伤后发病机理的研究,一直是肝病领域的热点。近年来,关于Hedgehog信号通路(hedgehog signaling pathway,HH)在肝脏疾病中的作用受到较多的关注[3-4]。HH由于存在天然植物阻断剂(藜芦类生物碱,环王巴明cyclopamine)[5-6],阐明其在肝损伤后反应过程中通讯作者：李淳,E-mail：lichunchris@hotmail.com的作用,寻找具有阻断作用的中药类似物,对开发中药新药具有重要的现实意义。, 百拇医药(蔡毅东 王丽 郑浩轩 孙淑美 王洋 李淳)
参见：首页 > 医疗版 > 疾病专题 > 胃肠外科 > 腹部创伤 > 肝损伤