槐定碱预防小鼠内毒素性肝损伤的作用及对肝脏CD14、TLR4表达的影响  

Effect of Sophoridine on Acute Liver Injury in Mice with Endotoxemia and its Impact on Expression of CD14 and TLR4

作　　者：王琳琳[1] 周娅[1] 梁锦屏[1] 黄菱[1] 高晓峰[1] 刘静[1] 王宁萍[1] 赵建宁[1] 

机构地区：[1]宁夏医科大学基础医学院病原生物学与免疫学教研室,银川750004

出　　处：《宁夏医科大学学报》2009年第6期709-712,722,F0003,共6页Journal of Ningxia Medical University

基　　金：国家自然科学基金(30660227);教育部新世纪优秀人才支持计划(NCET-06-0916);宁夏自然科学基金(NZ0783)

摘　　要：目的研究预防性给予槐定碱对小鼠内毒素血症急性肝损伤的影响及对CD14、TLR4表达的调节。方法观察槐定碱12、6、4mg/kg三个剂量预防性处理的小鼠,在给予内毒素后2、6、12、24h时各组小鼠一般状况、肉眼及光镜下肝组织的病理变化,赖氏法测定血清丙氨酸氨基转氨酶(ALT)浓度,RT-PCR与Western Blot分别检测肝组织CD14及TLR4的mRNA与蛋白表达,放免法检测血清TNF-α含量的变化。结果槐定碱三个剂量预防性给予,各个时间点结果均表明改善了内毒素血症模型小鼠的一般状况,减轻了肝组织病理改变;槐定碱三个剂量组的ALT值均低于同时间点的LPS模型组(P<0.01或P<0.05);槐定碱三个剂量组与同时间点LPS模型组比较,均下调肝组织CD14、TLR4 mRNA和蛋白的表达(P<0.01或P<0.05);槐定碱三个剂量组血清TNF-α水平均较同时间点LPS模型组降低(P<0.01或P<0.05),并在24h时均降至正常对照组水平。结论槐定碱12、6、4mg/kg三个剂量预防性给予,可抑制内毒素血症小鼠肝脏炎症反应,其机制可能与下调LPS识别受体CD14、TLR4表达,并进而抑制下游炎症因子分泌有关。Objective To observe the preventive effect of Sophoridine on liver inflammatory reaction of mice with endotoxemia,and its effect on the expression of CD14 and TLR4.Methods The ICR mice were administered by three dosage of Sophoridine, that was high doses （12mg/kg）, middle doses（6mg/kg）and low doses（4mg/kg）. 2, 6,12,24 hours after LPS injection, general symptoms were recorded,the blood and liver samples were taken and pathological damages of liver were checked. The level of alanine aminotransaminase（ALT） in serum was measured by Reitman mensurate rating. Reverse transcription polymerase chain reaction （RT-PCR） was employed to detect the mRNA expression of CD14 and TLR4 in liver. Protein expression of CD14 and TLR4 in liver were assessed by Western Blot. Serum TNF-a levels were determined by radio immunoassay. Results The results of three dosage of Sophoridine pretreated groups at each time-point showed that Sophoridine could ameliorate the general symptoms of endotoxic liver injury models significantly and decrease pathological damage of liver tissue. The three Sophoridine group with different doses showd a significantly lower ALT value than that in the LPS model at the same time point （P 〈 0.01 or P 〈 0.05）; compare to the LPS model, the express of CD14 , TLR4 mRNA and proteins in the liver of the three different doses sophoridine group were down-regulated significantly（ P 〈 0.01 or P 〈 0.05）, and the levels of serum TNF-a of three Sophoridine group were significantly reduced than those in the LPS model at the same time point（ P 〈 0.01 or P 〈 0.05）, and they reached the level of normal control groups at 24h. Conclusion The mechanism of Sophoridine anti-endotoxin liver injury had a strong relation with down-regulating of the expression of CD14 and TLR4, and influence of TLR4 transduction pathway, and restraining secretion of downstream inflammatory factors.

关 键 词：苦豆子 槐定碱 内毒素 肝脏 CD14 Toll样受体4 

分 类 号：R285.5[医药卫生—中药学]