目的探讨黑木耳提取物对脓毒血症大鼠血清中内毒素、肿瘤坏死因子(TNF-α)、白介素-1(IL-1)、白介素-6(IL-6)水平及肠、肝组织中核转录因子-κB(NF-κB)表达的影响。方法将健康Wistar大鼠随机分为对照组、实验组、蒙脱石散组、黑木耳多糖组、黑木耳粗提物组,术前分别预防性灌胃3d。第4天行手术,使用盲肠结扎穿刺法制备脓毒血症动物模型,造模12h后分别于门静脉和腹主动脉取血,检测血清内毒素水平;应用ELISA试剂盒检测血清中TNF-α、IL-1、IL-6水平;采用免疫组织化学法检测肝、肠组织细胞中NF-κB的表达水平。结果实验组、蒙脱石散组、黑木耳粗提物组及黑木耳多糖组门静脉血清和动脉血清内毒素水平均高于对照组,差异有统计学意义(P<0.05);黑木耳粗提物组及黑木耳多糖组门静脉血清和动脉血清内毒素水平均明显低于实验组及蒙脱石散组,差异有统计学意义(P<0.05)。实验组、蒙脱石散组、黑木耳粗提物组及黑木耳多糖组大鼠血清中TNF-α、IL-1、IL-6水平较对照组升高,差异有统计学意义(P<0.05);蒙脱石散组、黑木耳粗提物组及黑木耳多糖组大鼠血清中TNF-α、IL-1较实验组降低,差异有统计学意义(P<0.05)。实验组大鼠肝、肠组织细胞NF-κB强阳性表达,蒙脱石散组、黑木耳粗提物组和黑木耳多糖组肝、肠组织细胞NF-κB弱阳性表达,通过半定量测定,蒙脱石散组、黑木耳粗提物组和黑木耳多糖组肝、肠细胞NF-κB表达水平较实验组降低,差异有统计学意义(P<0.01)。结论黑木耳提取物在有效降低脓毒血症大鼠血清中内毒素的基础上,同时降低TNF-α、IL-1、IL-6含量,抑制脓毒血症大鼠肝、肠组织细胞NF-κB的表达。
Objective To discuss the effects of A .auricula-judae extract on rat model of sepsis ,this study mainly examines the serum concentration of endotoxin ,tumor necrosis factor (TNF-α) ,interleukin-1 (IL-1) , interleukin-6 (IL-6) and nuclear factor-κB (NF-κB) expression in intestinal and liver tissue .Methods 50 Wistar rats were randomized into 5 groups :sham surgery control group ,experiment group ,montmorillon-ite powder group ,A .auricula-judae polysaccharide group and crude A .auricula-judae extract group .Intra-gastric administration was performed every 8 hours three days prior to the surgery (1 .5 mL/100 g) .Sepsis models were induced by cecal ligation and puncture (CLP) procedure .Blood was collected from portal vein and abdominal aorta ,endotoxin concentration was detected by UV-Vis ;ELISA test was performed to de-termine the serum level of TNF-α,IL-1 ,IL-6 ;and immunohistochemistry was processed to detect NF-κB expression in intestinal and liver tissue .Results Two A .auricula-judae groups showed a much lower con-centration of serum endotoxin than the experiment group and montmorillonite powder group (P <0 .01);while the differences between the experiment group and montmorillonite group showed no statistical signif-icance (P >0 .05) .As to the three cytokines ,the montmorillonite powder group and two A .auricula-judae groups all presented lower level of the examined cytokines than the experiment group (P <0 .05);while the differences between montmorillonite powder and two A .auricula-judae groups showed no statistical sig-nificance (P >0 .05) .Unlike a weak staining detected in the control group ,experiment group showed a strong positive NF-κB staining (P < 0 .01);the staining in montmorillonite powder group was positive , and a moderate staining was detected in the two A .auricula-judae groups .Conclusion A .auricula-judae extract effectively decreases serum endotoxin concentration as well as serum level of TNF-α,IL-1 ,IL-6 in sepsis models ;it also prohibits the activation of NF-κB in intestinal and liver tissue of sepsis rats .