温下健脾法对脓毒症胃肠功能障碍大鼠肠道菌群及肠道免疫的影响

Effect of Warm Purgative and Strengthening Spleen Therapy on Intestinal Microecology and Immunity in Sepsis Rats with Gastrointestinal Dysfunction

目的观察温下健脾法对脓毒症胃肠功能障碍大鼠肠道菌群及肠道免疫的影响。方法24只SPF级SD雄性大鼠采用盲肠结扎穿孔法复制大鼠脓毒症模型成功后随机均分为模型对照组、中药组,并以10只SD大鼠行假手术作为正常对照组。术后各组大鼠均禁食不禁水饲养。中药组大鼠在术前及术后24h给予大黄附子汤加味中药(4mg/g,2次/24h)灌胃,正常对照组和模型对照组给予等量灭菌注射用水灌胃。造模成功24h后观察各组大鼠死亡率,留取存活大鼠留取粪便、血清及肠道组织,采用16SrDNA测序分析肠道菌群变化。采用ELISA方法检测肠道黏膜sIgA及血清D-乳酸的水平。结果脓毒症大鼠死亡率在中药组(16.7%)小于模型组(33.33%)(P<0.05)。在肠道菌群物种丰度方面,Bacteroidetes表达水平在模型对照组(24.36±7.15)<正常对照组(45.20±6.05)<中药组(49.69±7.98)(P<0.05)。D-乳酸水平在正常对照组(7.90±0.69)pg/mL<中药组(9.20±0.56)μg/mL<模型组(12.16±0.75)μg/mL(P<0.05),模型组大鼠肠道sIgA表达水平(11.57±0.31)μg/mL低于中药组(21.25±7.41)μg/mL和正常对照组(26.27±2.05)μg/mL(P<0.05)。结论温下健脾法可调节脓毒症胃肠功能障碍的肠道菌群及肠道免疫异常,从而发挥治疗脓毒症的作用。

Objective:To investigate the effect of warm purgative and strengthening spleen therapy(WPSST) on intestinal microecology and immunity in sepsis rats with gastrointestinal dysfunction. Methods:24 SPF SD rats were made into sepsis model with cecum ligation perforation method,and equally divided into the model group and the traditional Chinese medicine group. 10 SD rats with sham surgery were used as the control group. Rats in each group were raised with water feeding and without food. Rats in TCMG were administered by gavage of modified Dahuang Fuzi Decoction(4 mg/g of body weight,twice a day) using a tube 2 hour prior to CLP and after successful induction,rats in CG and MG by gavage of equivalent sterilized water. After 24 hours of administration,the mortality of rats in each group was recorded. The concentrations of D-lactic acid as well as sIgA in intestinal mucosa were evaluated with ELISA. The gut microbiota was detected by 16S rDNA gene sequencing. Results:The mortality rate of sepsis rats in TCMG(16.7%) was lower than that in MG(33.33%)(P < 0.05);the expression levels of Bacteroidetes:MG(24.36±7.15)<CG(45.20±6.05)<TCMG(49.69±7.98)(P < 0.05);the levels of D-lactic acid:CG(7.90±0.69) pg/mL<TCMG(9.20±0.56)μg/mL < MG(12.16±0.75)μg/mL(P < 0.05);the expression level of sIgA in MG[(11.57±0.31)μg/mL] was lower than that of CG[(26.27±2.05)μg/mL] and TCMG [(21.25±7.41)μg/mL](P < 0.05). Conclusion:WPSST can regulate intestinal flora and intestinal immune abnormalities in sepsis with gastrointestinal dysfunction,thus playing a role of treating sepsis.