血必净注射液对脓毒症大鼠血清内毒素及脾脏特异性免疫功能的影响

Effect of Xuebijing injection(血必净注射液) on level of serum endotoxin and specific immunological function of spleen in rats with sepsis

目的:观察脓毒症大鼠血中内毒素水平及脾淋巴细胞表面人白细胞DR抗原(HLA-DR)、脾淋巴细胞增殖及脾淋巴细胞天冬氨酸特异性半胱氨酸蛋白酶-3(caspase-3)特异性免疫功能状态的变化以及血必净注射液对脓毒症免疫功能的影响。方法:采用盲肠结扎穿孔术(CLP)制备脓毒症模型,将72只雄性SD大鼠按随机数字表法分为假手术组、模型组、左氧氟沙星组和血必净组。制模后3、24和72 h经腹腔静脉取血后活杀动物取脾脏;采用改良基质显色法检测血中内毒素水平,采用流式细胞仪检测HLA-DR水平,采用免疫组化法检测caspase-3阳性细胞表达,采用四甲基偶氮唑盐微量酶反应比色法(MTT)检测脾淋巴细胞的增殖。结果:模型组大鼠血中内毒素水平、脾脏淋巴细胞增殖反应和caspase-3阳性细胞表达指数均较假手术组明显升高,脾脏HLA-DR百分比较假手术组明显降低(P均<0.01)。血必净注射液能明显降低脓毒症大鼠血中内毒素水平,提高脾HLA-DR百分比,促进淋巴细胞增殖,降低脾淋巴细胞caspase-3阳性细胞表达,与模型组比较差异均有显著性,且作用优于左氧氟沙星组(P<0.05或P<0.01)。结论:脓毒症大鼠的免疫功能处于紊乱状态,特异性免疫功能受到抑制;血必净注射液能改善脓毒症时的特异性免疫功能状态。

Objective： To observe the characteristic changes of specific immunological function of serum endo toxin level, spleen lymphocyte human leukocyte antigen （locus） DR （HLA- DR）, limphopolesis and positive expression of caspase-3 in rats with sepsis and the effect of Xuebijing injection （血必净注射液） on their immunological function. Methods： Sepsis model was produced by cecal ligation and puncture （CLP）. Seventy -two male SD rats were randomly divided into four groups： sham operation, model, levofloxacin and Xuebijing injection. Blood samples were taken from the vein in the abdominal cavity at 3, 24 and 72 hours after the models were established, afterwards the rats were sacrificed and their spleens were taken out. Matrix development process was employed to examine the level of serum endotoxin, FACS flow cytometer was used to examine the expression level of HLA -DR, immunohistochemistry method was applied to examine the positive expression of caspase - 3 in spleen, and the method of methyl thiazolyl tetrazolium （MTT） was employed to examine the lymphopoiesis. Results： The levels of serum endotoxin, lymphopoiesis and caspase - 3 positive expression index in model group were obviously elevated than those of sham operation group, and percentage of HLA -DR was markedly lowered （all P〈 0. 01 ）. The results showed that Xuebingjing injection could significandy cut down the level of serum endotoxin and the positive expression of caspase - 3 in spleen lymphocyte, and it could markedly elevate the level of lymphopoiesis and percentage of HLA - DR. The differences were significant between Xuebijing injection group and those in model group, and the results in the former group were better than those in levofloxacin group （P〈0.05 or P〈0.01）. Conclusion： Disturbance of immunological function exists in the rats with sepsis, and the specific immunological function is under a state of suppression. Xuebijing injection may obviously improve the specific immunological function in rats with sepsis.