实验性肝炎大鼠肥大细胞的变化及c-kit和干细胞因子在肝组织的表达  

Changes in mast cells and hepatic expression of c-kit and stem cell factor in the rat model of chronic hepatitis

作　　者：李红[1] 赵龙凤[1] 郝彦琴[1] 尹镭[2] 赵元昌[2] 韩德五[2] 

机构地区：[1]山西医科大学第一医院感染病科,太原030001 [2]山西医科大学肝病研究所

出　　处：《中华肝脏病杂志》2013年第11期869-873,共5页Chinese Journal of Hepatology

基　　金：山西医科大学细胞生理学省部共建教育部重点实验室主任基金（2010-03）

摘　　要：目的研究肥大细胞（MC）对实验性肝炎大鼠的影响及其作用机制。方法取Wistar大鼠30只，随机分为2组：正常对照（NC）组、慢性肝炎（CH）组。CH组采用复合因素造模（用40％四氯化碳油溶液皮下注射，同时辅以低蛋白、低胆碱、高脂肪、高醇饮食）。4周末处死动物，取血分别检测血浆类胰蛋白酶（TS）、组织胺（HA）水平，同时观察肝脏组织学变化及Me形态改变。应用免疫组织化学方法观察肝脏c-kit和干细胞因子（SCF）的表达。用逆转录一多聚酶链反应（RT-PCR）方法观察肝组织c-kitmRNA和SCFmRNA的表达水平。组间比较采用SPSS统计学软件进行f检验，相关分析采用直线相关回归分析，P〈0．05为差异具有统计学意义。结果（1）与正常对照组比较，CH组血和肝组织HA水平[（78．09±38．55）ng／对比（145．14土52．54）ng／ml和每100mg肝组织中（51．58±20．45）ng对比（106．59±43．15）rig，f值为2．457和2．825，P〈0．051，以及TS均有明显升高[（0．476士0．143）mg／ml对比（0．753±0．210）mg／ml，f值为4．165，P〈0．051。（2）光镜下，CH组有脂肪变性和纤维化形成；甲苯胺蓝染色可见CH组肝脏血管周围及纤维间隔内7kit正在脱颗粒和已经脱颗粒的充满紫色颗粒的MC。CH组Me数目较正常对照组明显升高[（2．167±0．924）个／lap对比（10．92±1．575）个／hp，t=7．633，P〈0．051。（3）免疫组织化学染色结果显示CH组高表达c-kit和SCF（相对表达量2．783±0．577对比12．86土3．126和3．383±1．583对比15．58±6．431，t值分别为9．511和9．625，P〈0．05），而且二者的表达均与HA水平呈明显正相关（，值分别为0．478、0．556，JP〈0．05）。RT-PCR结果显示CH组c-kitmRNA和SCFmRNA表达水平明显增高。结论CH时Me脱颗粒明显增加，HA释放增加，可能与膜受体c-kit及其配体SCF的表达增加有关。Objective To study the potential role of mast cells and the related molecular mechanism in chronic hepatitis （CH） using a rat model system. Methods Thirty Wistar rats （15 males, 15 females; weight range： 230-290 g） were randomly divided into the normal contrast （NC） group and experimental CH group. The CH group received subcutaneous injection of CC14 and a diet high in cholesterol and alcohol content and low in protein and choline content. Throughout the 4-week modeling period, aseptic blood samples were taken to test plasma tryptase （TS） and hyaluronic acid （HA） levels. The rats were euthanized to assess the changes in liver mast cells by histology and morphology analyses and the changes in liver expression of c-kit and stem cell factor （SCF） proteins by immunohistochemistry and mRNAs by RT- PCR. Results Compared to the NC group, the CH group had higher plasma and liver concentration of HA （78.09 ±38.55 vs. 145.14±52.54 ng/ml, 51.58 ±20.45 vs. 106.59 ±43.15 ng/100 rag; t = 2.457 and 2.825respectively, both P 〈 0.05） and TS （0.416 ± 0.143 vs 0.753 ± 0.210 mg/ml; t = 4.165, P 〈 0.05）. The CH group also showed fatty degeneration and fibrosis with many degranulating and degranulated mast cells filled with purple granula located around the liver blood vessels and in fiber-intervals. The CH livers also showed a significantly higher number of mast cells （2.167 ± 0.924 vs. NC： 10.92 ± 1.575; t = 7.633, P 〈 0.05） and stronger intensity of c-kit staining （2.783 ± 0.577 vs. 12.86 ± 3.126; t = 9.511, P 〈 0.05） and SCF staining （3.383 ±1.583 vs. 15.58 ±6.431; t = 9.625, P 〈 0.05）. The expressions of c-kit and SCF were positively correlated with HA level （r = 0.478 and 0.556 respectively, both P 〈 0.05）. The c-kit and SCF mRNA expression levels were also significantly higher in the CH liver tissues. Conclusion Mast cell degranulation and histamine release is significantly increased under conditions of chronic hepatitis, and the related mechanism may i

关 键 词：肝炎 大鼠 肥大细胞 C-KIT 干细胞因子 

分 类 号：R575.1[医药卫生—消化系统]