菌毒炎并治对脓毒症大鼠模型影响的肝基因芯片研究

A gene chip study of ＂Jun Du Yan Bingzhi＂ on liver in a sepsis model of rat

目的应用基因芯片技术探讨菌毒炎并治对脓毒症大鼠基因的改变，以期从基因水平上探讨其药物作用机制。方法将90只大鼠随机分为正常对照组、模型组、菌毒炎组，每组30只；采用盲肠结扎穿孔术（CLP）复制脓毒症模型。菌毒炎组于制模后腹腔注射泰能0．18g／kg、血必净注射液10ml／kg、中药复方凉膈散煎剂15ml／kg（药物浓度6kg／I，）灌胃，每日2次；正常对照组和模型组腹腔给予生理盐水10ml／kg。观察大鼠存活时间及48h和72h的存活率；应用BiostarR-40S芯片检测模型组／正常对照组，菌毒炎组／模型组鼠肝基因，并以Cy3和Cy5二者荧光信号相对强度比值≥2．0或≤0．5筛选差异显著基因，通过美国国立生物技术信息中心（NCBI）数据库查询基因功能并加以分类。结果菌毒炎组大鼠48h和72h存活率明显高于模型组（83．3％比30．0％，76．7％比17．7％，P均〈0．01），模型组／正常对照组共筛选出305个差异基因，其中上调159个、已知基因109个，下调基因146个、已知基因89个；菌毒炎组／模型组共筛选出500个差异基因，其中上调292个、已知基因175个，下调基因208个、已知基因148个。模型组／正常对照组上调而菌毒炎组／模型组下调的已知基因共有48个，模型组／正常对照组下调而菌毒炎组／模型组上调的已知基因共有63个。结论菌毒炎并治可显著降低脓毒症大鼠48h和72h死亡率；通过调控免疫反应、炎症、信号转导、转录调节、细胞周期、细胞凋亡、新陈代谢、蛋白翻译／加工／修饰／降解、细胞分化／增殖／生长等相关基因，能促使脓毒症大鼠多系统功能恢复正常。

Objective To study the effect of ＂Jun Du Yan Bingzhi＂ on genie change in liver of a sepsis rat model by gene chip technique, in order to study the mechanism of the action of the drug on the gene level. Methods Ninety rats were randomly divided into normal control group, model group and ＂ Jun Du Yan Bingzhi＂ group, with 30 rats in each group. Sepsis was reproduced by cecal ligation and puncture （CLP） method. In ＂Jun Du Yan Bingzhi＂ group the rats were treated with intraperitoneal injection of imipenem/ cilastatin （0.18 g/kg）, Xuebijing injection （10 ml/kg） and gavage of ＂Liangge San＂ （15 ml/kg）. In the control group and model group intraperitoneal physiological saline （10 ml/kg） was given. Survival time, and 48-hour and 72-hour survival rates of every group were observed, and changes in liver genes were examined with BiostarR-40 s chip. The ratio of Cy3/Cy5 ≥2.0 or ≤0.5 was used to screen differential genes, and NCBI database was used to identity the function of differential genes. Results The 48-hour and 72-hour survival rate of ＂Jun Du Yan Bingzhi＂ group was significantly higher than that of model group （83.3% vs. 30.0%, 76.7% vs. 17.7%, both P〈0.01）, 305 differential genes were found in model/crotrol groups, with up-regulation in 159, down-regulation in 146, 500 differential genes were found in ＂ Jun Du Yan Bingzhi＂ group/model group, with up-regulation in 292, down-regulation in 208, model group/control group up-regulation and ＂Jun Du Yan Bingzhi＂ group/model group down-regulation were 48, model group/control group down regulation and ＂Jun Du Yan Bingzhi＂ group/model group up-regulation were 63. Conclusion ＂Jun Du Yan Bingzhi＂ can degrade the 48-hour and 72-hour death rate of sepsis rat, through control immunization related, inflammation, signal transduetion, transcription regulation, cell cycle, apoptosis, substance metabolism, translation/processing/modify/degradation of protein, differentiation/proliferation/ growth of cell related gene, promote multisystem function of sepsis rat to recover normal.