甘氨酸抑制脂多糖介导的鼠枯否细胞激活的时相选择及机制探讨  

The mechanism and treatment phases chosen of glycine for inhibition lipopolysacharide induced Kupffer cells activation

作　　者：刘作金[1] 游海波[1] 李旭宏[1] 陈先锋[1] 刘海忠[1] 彭勇[1] 刘长安[1] 龚建平[1] 

机构地区：[1]重庆医科大学附属第二医院肝胆外科重庆市肝胆外科重点实验室,400010

出　　处：《中华外科杂志》2006年第3期189-192,共4页Chinese Journal of Surgery

基　　金：国家自然科学基金资助项目(30471969;30500473);重庆市自然科学基金资助项目(2005BB5242);中国高等学校博士学科点专项科研基金资助项目(2005063004601)

摘　　要：目的探讨甘氨酸抑制脂多糖介导的鼠枯否细胞激活效应相关机制和甘氨酸的最佳用药时机。方法将40只BALB/c小鼠分为内毒素组、预防组、早期治疗组和后期治疗组,每组10只。分离培养枯否细胞后,内毒素组加入脂多糖(10mg/L),预防组、早期治疗组和后期治疗组分别在加入脂多糖前24h、加入后0和4h加入甘氨酸(1mmol/L),分别在加入脂多糖后0、1、2、6和12h,采用逆转录聚合酶链反应及蛋白印迹法测定枯否细胞的白细胞介素1受体相关激酶4(IRAK4)mRNA和蛋白表达水平,用酶联免疫吸附法检测枯否细胞的核因子κB(NFκB)活性和培养上清液的肿瘤坏死因子α(TNFα)含量。结果脂多糖刺激后,预防组的IRAK4mRNA和蛋白表达、NFκB活性的相对峰值分别为3.64±1.13、34.54±10.31、0.47±0.10,TNFα峰值为(1780.70±210.17)pg/ml,与早期治疗组比较,差异均无统计学意义,但与内毒素组和后期治疗组比较,各峰值均明显降低,差异有统计学意义。结论提前或者在脂多糖刺激的同时应用甘氨酸,能有效抑制脂多糖介导的枯否细胞激活效应,其作用机制之一可能为抑制IRAK4的表达。Objective To explore the possible mechanism and optimal treatment phase of glycine for inhibition lipopolysaccharide （LPS） induced Kupffer cells （KCs） activation. Methods The KCs were isolated from 40 BALB/c mice and divided into four groups： the endotoxin group, the prevention group, the early treatment group and the later treatment group （n = 10）. The endotoxin group was treated with 10 mg/L LPS, and in the other three groups, glycine （ 1 mmol/L） was given 24 h before, or at 0 h or 4 h respectively after LPS stimulation. At 0 h, 1 h, 2 h, 6 h and 12 h after LPS stimulation, the mRNA levels and protein expression of interleukin-1 receptor associated kinase-4 （IRAK-4） were determined by reverse transcription polymerase chain reaction （RT-PCR） and Western blot respectively, and nuclear factor-kappaB （NF-κB） activities as well as tumor necrosis factor alpha （TNF-α） levels were also detected by enzyme-linked immunosorbent assay （ELISA）. Results The climax values of IRAK-4, NF-κB and TNF-α were significantly higher in the endotoxin group and the later treatment group than that in the other two groups （t=3.17, 4.33, 2.47, 126.73, P 〈0.01）. Conclusion The results indicated that prophylactic or simultaneous treatment with glycine could effectively inhibit LPS-induced KCs activation by inhibiting IRAK-4 expression.

关 键 词：枯否氏细胞 甘氨酸 内毒素类 受体 白细胞介素1 

分 类 号：R96[医药卫生—药理学]