大鼠肠巨噬细胞TNFα表达及复方大承气汤的影响

TNFαexpression and effects of Dachengqi Decoctionin compound in gut macrophages

目的:探讨体外培养的肠巨噬细胞分泌TNFα的规律及地塞米松、TNFα单克隆抗体及复方大承气汤对LPS诱导的肠巨噬细胞TNFα产生的影响。方法:体外分离培养大鼠肠巨噬细胞。设对照组、脂多糖组、地塞米松处理组、TNFα单抗处理组和复方大承气汤处理组.每组分别在3h,6h,12h和24h取上清液,用放免法检测TNFα浓度。并分别收集肠巨噬细胞,提取RNA,采用RT-PCR法相对定量TNFα mRNA。结果:肠巨噬细胞经脂多糖(LPS)诱导后,各时相TNFα分泌及TNFmRNA表达均明显增加;地塞米松、TNFα单克隆抗体及复方大承气汤处理后,各时相TNFα分泌都较脂多糖(LPS)诱导组显著下降,各处理组TNFα mRNA表达在3h时与脂多糖(LPS)诱导组比较无明显差异,6h、12h、24h,地塞米松、复方大承气汤处理组显著降低,而TNFα单克隆抗体处理组无明显差异。结论:LPS是肠巨噬细胞分泌TNFα的有效激活剂,地塞米松、复方大承气汤能从蛋白质及核酸水平抑制TNFα的产生,而TNFα单克隆抗体只能从蛋白质水平抑制TNFα的产生。

AIM:To explore the rule of producing TNFα in gut macrophages through respects of gene expression and protein synthesis of TNF α and to observe the effect of dexamethasone (DEX), monocolonal antibody to tumor necrosis factor alpha (TNF α moAb) and Fufang Dachengqi Decoction compound on TNF α production. METHODS:The cultured rat gut macrophages were divided into five groups: control group (group C), lipopolysaccharide (LPS) group (group L), DEX treated group (group L+D), TNF α-MoAb treated group (group L+M), LPS plus Fufang Da Chengqi Decoction treated group (group L+F). Each group was divided into four phases: cultured for 3, 6, 12, and 24 h. The supernatants were collected and frozen at-70℃until TNFα was determined, and the cells were used to isolate the RNA. TNFα levels were determined by radioimmunoassay method. The expression of TNFα mRNA was evaluated by RT-PCR. RESULTS:The level of TNFα and expression of TNFα mRNA significantly increased at each phase in group L.The level of TNFα significantly decreased at each phase in group L+D, group L+M and group L+F compared with group L. The expression of TNFα mRNA in each treatment group had no obvious difference, compared with group L in 3h, but group L+D and group L+F significantly decreased in 6, 12, and 24h. Group L+M still showed no obvious difference. CONCLUSION:Gut macrophage induced by LPS can produce more TNFα through its gene expression and protein synthesis. DEX, Fufang Dachengqi Decoction can suppress the TNFα mRNA transcription and protein synthesis of TNFα; TNFα-moAb only lowered the level of TNFα protein. The three drugs have protective effects in inflammatory mediators reaction, gut barrier damage and multiple organ dysfunction syndrome (MODS).