免疫调节治疗对脓毒症模型大鼠免疫失衡状态及生存时间的影响

Effects of immune regulation therapy on immune imbalance and survival time in septic rats

目的观察应用血必净注射液作为免疫调节治疗对脓毒症模型大鼠免疫失衡状态及生存时间的影响。方法按随机数字表法将180只雄性SD大鼠分为假手术组(A组,n=15)、脓毒症模型组(n=165),脓毒症模型组采用盲肠结扎打孔(cecal ligation and puncture,CLP)制备脓毒症模型。建模成功后,剔除死亡大鼠,脓毒症模型组再分为对照组(B组,n=54)、治疗组(C组,n=53)、试验组(D组,n=54)三个亚组。A组及B组术后均皮下注射生理盐水。C组在B组治疗的基础上应用0.2%盐酸左氧氟沙星注射液,D组在C组治疗基础上应用血必净注射液。分别于6 h、24 h、48 h、72 h存活大鼠眼内眦静脉取血,检测血清白细胞介素-6(IL-6)水平及CD14+单核细胞人白细胞抗原-DR(human leucocyte antigen-DR,HLA-DR)表达率,记录各大鼠的存活时间。结果所有CLP大鼠全程血清IL-6水平均明显升高(P<0.05),B组于6 h即明显升高,24 h达峰值,随后下降,C组、D组趋势表现与B组相同,但升高的幅度低于B组。抗感染治疗48 h即可使C组血清IL-6水平较B组降低[IL-6(pg/mL):190.95±28.77 vs.262.09±49.66,P<0.05],应用血必净后24 h即可使D组IL-6较B组降低[IL-6(pg/mL):228.86±32.08 vs.270.65±52.21,P<0.05],48 h可使D组IL-6较C组降低[IL-6(pg/mL):157.52±26.11 vs.190.95±28.77,P<0.05]。6 h、24 h时所有CLP大鼠HLA-DR表达率均出现减低(P<0.05),后逐渐上升,72 h时最终存活的各组大鼠HLA-DR表达率均恢复正常(P>0.05);6 h、24 h时抗感染治疗后可使C组HLA-DR表达率较B组略低的大鼠存活(P>0.05),血必净可使D组HLA-DR表达率较C组更低的大鼠存活[HLA-DR(%):47.78±15.49 vs.59.02±17.38(6 h),55.24±14.94 vs.65.32±15.44(24 h),均P<0.05];72 h时抗感染治疗可使C组较B组生存率升高及生存时间延长[生存率(%):28.30 vs.11.11,生存时间中位数(h)58 vs.47(24 h),均P<0.05];应用血必净注射液后D组较C组生存率提高,生存时间延长[生存率(%):50.00 vs.28.30,生存时间中位数(h)69 vs.58(24 h),均P<0.05]。结论脓毒症可导致免疫失衡,血必净注射液可减轻促炎反应及免疫麻痹,减轻炎症反应,延长生存时间。

Objective To observe the effects of Xuebijing used as immunomodulatory therapy on immune imbalance and survival time in septic rats.Methods 180 male SD rats were divided into sham operated group(group A,15 rats)and septic group(165 rats).The septic rats were established by cecal ligation and puncture(CLP).The survival septic rats were randomized into three groups:control group(group B,54 rats),treatment group(group C,53 rats),and experimental group(group D,54 rats).Group A and group B animals were injected subcutaneously with normal saline.Group C animals were injected with levofloxacin and physiological saline.Group D animals were injected with Xuebijing,levofloxacin and physiological saline.Blood was collected from the vein of canthus of the surviving rats at 6,24,48 and 72 h,respectively.The expression levels of interleukin-6(IL-6)and CD14+monocyte human leukocyte antigen-DR(HLA-DR)were measured,and the survival time(h)of each rat was recorded.Results The level of IL-6 in all CLP rats increased significantly(P<0.05).Group B increased significantly at 6 h,peaked at 24 h,and then decreased.The trend of groups C and D was the same as group B,but the increase was lower than that in group B.After administration of levofloxacin for 48 hours,the level of IL-6 in group C was significantly lower than that in group B[IL-6(pg/mL):190.95±28.77 vs.262.09±49.66,P<0.05].After administration of Xuebijing for 24 hours,the level of IL-6 in group D was lower than that in group B[IL-6(pg/mL):228.86±32.08 vs.270.65±52.21,P<0.05],and the level of IL-6 in group D after Xuebijing treatment for 48 hours was lower than that in group C[IL-6(pg/mL):157.52±26.11 vs.190.95±28.77,P<0.05].The HLA-DR expression rate of all CLP rats decreased at 6 h and 24 h(P<0.05),and then increased gradually.At 72 h,the expression rate of HLA-DR in the survival groups returned to normal(P>0.05);Anti-infection treatment for 6 and 24 hours could make the rats in group C survive(P>0.05),and Xuebijing could make the rats with lower expression rate of HLA-DR in group D survive compared with group C[HLA-DR(%):47.78±15.49 vs.59.02±17.38(6 h),55.24±14.94 vs.65.32±15.44(24 h),all P<0.05].Anti-infective therapy for 72 hours could improve the survival rate and prolong the survival time of group C compared with group B[survival rate(%):28.30 vs.11.11,median survival time(h)58 vs.47(24 h),all P<0.05].After Xuebijing was applied,the survival rate of group D was improved and the survival time was prolonged[survival rate(%):50.00 vs.28.30,median survival time(h)69 vs.58(24 h),all P<0.05].Conclusion Sepsis can cause immune imbalance;Xuebijing can alleviate pro-inflammatory response and immune paralysis,which can reduce the inflammatory reaction and ultimately prolong the survival time of rats.