姜黄素抑制内毒素所致肾脏炎症的作用基因表达谱的分析研究

Analysis of Gene Expression Profile in the Kidney of Curcumin on the Endotoxin-Induced Renal Inflammation

目的:肾脏的急慢性炎症与肾脏疾病的进展密切相关,本研究拟通过基因芯片技术及病理学方法探讨姜黄素对内毒素脂多糖(LPS)所致肾脏炎症的保护作用的机制。方法:SPF级昆明种小鼠9只,鼠龄6～8周,体重20～25 g。小鼠分为3组:(1)正常对照组:腹腔注射生理盐水;(2)模型组(LPS组):腹腔注射LPS(5 mg/kg);(3)治疗组(姜黄素+LPS):动物先腹腔注射姜黄素(5 mg/kg)3 d,然后腹腔注射LPS 5 mg/kg。于处理后6 h取材,切取部分肾组织用10%甲醛固定,HE,PAS染色及免疫组化,进行病理学观察;利用Affymetrix公司生产的mouse 430基因芯片分别检测正常对照组、模型组、姜黄素治疗组肾组织基因表达谱,并利用生物信息学方法对检测结果进行分析。结果:肾组织基因表达谱显示,符合LPS刺激后基因表达水平上升,并经姜黄素治疗后表达水平下降的共有148个Affy IDs(A组),而呈相反趋势的有133个Affy IDs(B组)。在前组的差异基因中,统计筛选得21个Gene Ontology(GO)的功能节点(P≤0.01)。在生物过程角度方面,其中富集量最高的为调节巨噬细胞活化和巨噬细胞活化相关基因。在细胞定位角度方面,有4个GO的功能节点(P≤0.01),在分子结构角度方面,有7个GO的功能节点(P≤0.01)。在B组中,在生物学过程角度方面,有2个GO的功能节点(P≤0.01);在细胞定位角度方面,有1个GO的功能节点(P≤0.01)。研究还发现了姜黄素抑制内毒素所致肾脏炎症作用机制的信号通路及可能的靶位基因。结论:利用基因芯片技术在阐明了炎症状态下肾脏基因的表达谱同时,证实姜黄素治疗可以改变其基因的表达谱,为进一步研究肾脏炎症纤维化的分子发病机制和姜黄素治疗的作用机制创造了条件。

Objective：Acute and chronic kidney inflammation are closely related to the progress of kidney diseases.In this study,we explored the protective mechanism of curcumin on lipopolysaccharide（LPS）-induced kidney inflammation by gene chip and pathological technology.Methods：Nine SPF Kunming mice（aged from 6 to 8 weeks,weight 20～25 g） were divided into three groups.Saline and LPS were injected by intraperitoneal in normal control group and model group,respectively.Mice in treatment group were first injected curcumin（5 mg/kg） for three days,then injected LPS（5 mg/kg）.Kidney tissues were harvested at 6 h after treatment,parts of kidney were fixed with 10% formaldehyde for HE,PAS staining and immunohistochemistry.Affymetrix gene chips（mouse 430 chip） were used to detect the renal gene expression profile,and the results were analyzed by bioinformatics methods.Results：Renal gene expression profile showed that there are 148 Affy IDs（A group） that the levels of gene expression were increased after LPS stimulation and decreased by the curcumin treatment,while there are 133 Affy IDs（B group） with the opposite trend.In the differentially expressed genes of A group,21 GO genes were selected by screening function（P≤ 0.01）.In the biological processes,most genes are related to the genes of regulation of macrophage activation and macrophage activation associated genes.In the cellular localization,there are four functional GO genes（P≤0.01）;in the molecular structure,there are 7 functional GO genes（P≤0.01）.In the B group,there are two functions of GO genes（P≤ 0.01） in the biological process;there is a function of GO genes（P≤0.01） in the cellular localization.In this study,we also found that the signaling pathway and the possible target gene of the protective effects of curcumin on endotoxin-induced renal inflammatory.Conclusion：The kidney gene expression profile in the inflammatory state was clarified by using gene chip technology.We also confirmed that curcumin treatment can change their gene expression profile.