左归丸对同种异性骨髓移植小鼠肝再生相关基因信号通路的影响  

Effects of Zuoguiwan on the liver regeneration related gene signaling pathway in female mice with male mice bone marrow transplant

作　　者：李瀚旻[1] 桂文甲[1] 李晶津[2] 高翔[1] 晏雪生[1] 程湧[2] 

机构地区：[1]湖北中医学院附属医院肝病研究所,湖北省武汉市430061 [2]武汉大学医学院2003级临床医学系,湖北省武汉市430072

出　　处：《中国组织工程研究与临床康复》2008年第31期6069-6073,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金重大研究计划项目(90709041);国家自然科学基金(30672590;30271562;30371787);国家重点基础研究发展计划(973计划)项目(2002CCC00300);湖北省自然科学基金(2001ABB171)~~

摘　　要：背景:维持正常的肝再生是避免肝纤维化和肝癌发生发展的关键环节,中医药调控肝再生主要反映在单味或复方中药对肝细胞、间质细胞、骨髓干细胞的增殖调控。目的:利用基因芯片技术探讨中药左归丸对同种异性骨髓移植小鼠肝再生相关基因信号通路的影响。设计、时间及地点:开放性实验,于2002-01/2007-12在湖北中医学院附属医院肝病研究所完成。材料:SPF级4周龄BABL/C小鼠,雄性10只,作为供体;雌性80只,作为受体,随机分为单纯骨髓移植组、左归丸治疗组,40只/组。小鼠16Kv1.0基因表达谱寡核苷酸芯片由上海生物芯片公司提供。左归丸由熟地、淮山药、枸杞子、山茱萸、菟丝子、川牛膝、鹿角胶、龟板胶组成。方法:两组受体小鼠接受60Co源γ射线照射后,经尾静脉输入供鼠单个核细胞悬液0.2mL(约2×106个细胞),移植后分别给予生理盐水、左归丸7g/(kg·d)灌胃,6个月后取肝组织标本。选用小鼠基因表达谱寡核苷酸芯片,提取液氮冻存肝组织总RNA,利用反转录酶合成掺有荧光标记的cDNA探针,将探针与基因表达谱芯片杂交,筛选样本之间杂交信号比值有差异表达的基因。利用GenBankID和UniGene数据库将差异表达基因聚类,按基因本体分类法对聚类的差异表达基因分类,并采用KEGG数据库查询差异表达基因所涉及的信号通路。主要观察指标:差异表达基因谱。与肝再生相关的代谢及信号通路。结果:①与单纯骨髓移植组相比,左归丸治疗组差异表达基因共1147条,已知功能基因有533条,其中209条基因涉及70种生物化学通路。②在差异表达基因涉及的代谢信号通路中,与肝再生相关的信号通路主要有Wnt信号通路、MAPK信号通路、TGFβ信号通路、Jak-STAT信号通路、凋亡、Toll样受体信号通路等,这些通路中Wnt1、EGF、FGF2、FGF16、MAPKK1、E2F、CSF3、Myd88、sFRP1、sFRP5、CSF2受体、CNTF受体、Caspase1BACKGROUND： Normal liver regeneration is key to avoid hepatic fibrosis and liver cancer incidence and progression. Single or compound traditional Chinese medicine can regulate the proliferation of hepatocyte, mesenchymal cells, and bone marrow stem cells. OBJECTIVE： To explore the effects of Chinese herb Zuoguiwan on the liver regeneration related gene signaling pathway in female mice with male mice bone marrow transplantation by gene array technique. DESIGN, TIME AND SETTING： The open experiment was performed at Institute of Liver Disease, Hospital of Hubei College of Traditional Chinese Medicine from January 2002 to December 2007 MATERIALS： Ninety SPF 4-week-old BABL/C mice, 10 males as donor and 80 females as recipients, were randomly divided into bone marrow transplantation group and Zuoguiwan group （n=40）. Mouse 16K v 1.0 oligonucleotide microarrays for gene expression profile were provided by Shanghai Biochip Co., Ltd.; Zuoguiwan was composed of Shudi, Huaishanyao, Gouqizi, Shanzhuyu, Tusizi, Chuanniuxi, Lujiaojiao, and Guibanfiao. METHODS： After exposed to irradiation with 9.0 Gy ^60Co Y -rays, the recipient mice were injected via the tail vein with the bone marrow cell suspension （0.2 mL, about 2×10^6 cells） of homogenic donor mice. Normal saline and Zuoguiwan （7 g/kg per day） were respectively intragastrically fused to the two groups after bone marrow transplantation. After 6 months of treatment, all mice were sacrificed to harvest liver tissues. Mouse oligonucleotide microarrays for gene expression profile was used to extract total RNA from the liver tissue that was preserved in liquid nitrogen to synthesize the probe containing fluorescence-labeled cDNA using the reverse transcription enzyme to synthesis. The probe and the gene expression profile microarray were hybridized to observe the change of the liver issues gene expression. Genes that had significant differences in hybrid signal ratio were selected. The differentially expressed genes were clustered using GenBankID an

关 键 词：骨髓移植 肝再生 左归丸 基因表达谱 信号通路 

分 类 号：R457.7[医药卫生—治疗学]