血必净注射液对重症脓毒症患者Toll样受体4及下游细胞因子的影响

A study on Effects of Xuebijing Injection on Toll-like Receptor 4 and Cytokines in Severe Sepsis Patients

目的:探讨血必净注射液对重症脓毒症患者Toll样受体4(TLR4)及下游细胞因子的影响。方法:将40例严重脓毒症患者随机分为常规治疗组(20例)和血必净治疗组(20例);以同期20例健康志愿者作为健康对照组。常规治疗组按严重脓毒症治疗指南进行治疗;血必净治疗组在常规治疗组的基础上加用血必净注射液100mL静脉滴注,q12h,连续7天。健康对照组于清晨取血一次。常规治疗组与血必净治疗组于治疗前、治疗后第1、3、7天清晨取血。采用酶联免疫吸附法(ELISA)测定血清白细胞介素6(IL-6)、肿瘤坏死因子-α(TNF-α),流式细胞仪检测单核细胞膜表面TLR4蛋白表达,半定量逆转录-聚合酶链式反应法(RT-PCR)检测单核细胞TLR4 mRNA表达。结果两组患者之间治疗前IL-6、TNF-a、TLR4蛋白和TLR4mRNA的表达水平比较,差异无统计学意义(P>0.05),但均明显高于健康对照组,差异有统计学意义(P<0.01)。两组患者治疗后IL-6、TNF-a、TLR4蛋白和TLR4mRNA的表达水平与本组治疗前比较均明显下降,差异有统计学意义(P<0.05或P<0.01),且血必净治疗组下降幅度大于常规治疗组,差异有统计学意义(P<0.05或P<0.01)。结论:重症脓毒症患者IL-6、TNF-a、TLR4蛋白和TLR4mRNA的表达水平均升高;血必净可能通过下调TRL4蛋白及TRL4mRNA水平抑制下游细胞因子的释放。

Objective：To investigate effects of Xuebijing injection on toll-like receptor 4 and cytokines in severe sepsis patients.Methods：Forty patients with severe sepsis were randomly divided into routine group and Xuebijing-treated group.Twenty healthy volunteers were taken as the normal control group.The routine group was treated according to the therapy guideline of severe sepsis.Beside the treatment used in the routine group,100ml Xuebijing injection was added by intravenous drip once every 12 hours for 7 days in the Xuebijing-treated group.The venous blood of the normal control group was collected once in the morning.Before treatment and 1,3,and 7 days after treatment,the venous blood of the routine group and the Xuebijing-treated group were collected.The expressions of interleukin-6（IL-6） and tumor necrosis factor-α（TNF-α）were measured by enzyme linked immunoadsorbent assay（ELISA）.The mRNA expression of Toll-like receptor 4（TLR4） of peripheral blood mononuclear cell（PBMC） was detected by semiquantitative reverse transcription-polymerase chain reaction（RT-PCR）.The protein expression of TLR4 of PBMC was detected by flow cytometry.Results：Before treatment,the expressions of IL-6,TNF-a,TLR4 protein and TLR4 mRNA in the Xuebijing-treated group showed no statistical significance with those in the routine group（P0.05）,and the above parameters in the two treatment groups were significantly higher than those in the normal control group（P0.01）.The above parameters in the two treatment groups were decreased significantly after treatment（P0.05 or P0.01）.All parameters at the same time points in the Xuebijing-treated group were lower than those in the routine group（P0.05 or P0.01）.Conclusion：The expressions of IL-6,TNF-a,TLR4 protein and TLR4 mRNA were upregulated markedly in severe sepsis patients.Xuebijing injection can effectively decreace cytokines.The mechanism is probably related to the decreace of TLR4 protein and TLR4 mRNA expressions.