炎调方对脓毒症大鼠肠道微生态环境的影响研究

Effect of Yantiao Decoction on Intestinal Microecological Environment of Sepsis Rats

目的观察脓毒症大鼠肠道微生态环境的变化以及炎调方对其的影响。方法将40只SD大鼠随机分为假手术组、模型组、炎调方组、益生菌组、联合组。模型组、炎调方组、益生菌组、联合组分别以生理盐水、炎调方、培菲康散、炎调方+培菲康散灌胃,连续3d。采用盲肠结扎穿孔术(CLP)制备脓毒症模型。假手术组以生理盐水灌胃,采用假手术法。造模24h后采集血清及粪便标本。采用ELISA法检测炎症因子白细胞介素-8(IL-8)、白细胞介素-10(IL-10)、肿瘤坏死因子-α(TNF-α)水平;通过Illumina MiSe q平台对粪便标本进行16Sr DNA测序。结果1)炎症因子比较:相较假手术组,模型组IL-8、TNF-α升高,IL-10下降(P<0.01);相较模型组,炎调方组、益生菌组和联合组IL-8、TNF-α水平减低,IL-10水平升高(P<0.01)。2)α多样性比较:相较假手术组,模型组simpson指数升高(P<0.01);相较模型组,炎调方组sobs指数、shannon指数升高(P<0.05),simpson指数减低(P<0.01)。3)物种差异比较:模型组黄杆菌科、理岩菌科和葡萄球菌科呈高丰度(P<0.05),疣微菌门呈低丰度(P<0.05);益生菌组双歧杆菌呈高丰度(P<0.001);炎调方组疣微菌门呈高丰度(P<0.05)。结论炎调方在调控脓毒症大鼠炎症因子水平,减轻过度的炎症反应的同时,可以提高脓毒症大鼠肠道菌群多样性和丰富度。

Objective:To observe the changes of intestinal microecological environment in sepsis rats and the effect of Yantiao Decoction on it.Methods:A total of 40 SPF SD rats were equally divided into the sham-operation group(J),model group(M),Yantiao Decoction group(Y),probiotics group(X)and Yantiao Decoction+probiotics group(H).Group J was given normal saline,and sham operation was performed.Group M,Group Y,Group X and Group H were given normal saline,Yantiao Decoction,Peifeikang San,and Yantiao Decoction+Peifeikang San respectively,for 3 days.Group J was given saline,and sham operation was performed.Serum and feces samples were collected 24 hours after modeling.Serum levels of IL-8,IL-10 and TNF-α were tested by ELISA.16 Sr DNA of the feces samples were tested by Illumina MiSeq platform.Results:1)Comparison of inflammatory cytokines:Compared with group J,IL-8 and TNF-α were increased and IL-10 was decreased in group M(P<0.01).Compared with group M,the levels of IL-8 and TNF-α in groups Y,X and H were decreased,the levels of IL-10 were increased(P<0.01).2)Comparison ofαdiversity:Simpson index of group M was higher than that of group J(P<0.01).Compared with group M,Sobs index and Shannon index in group Y were increased(P<0.05),while Simpson index was decreased(P<0.01).3)Species difference analysis:Flavobacteriaceae,Rikenellaceae and Staphylococcaceae showed high abundance in Group M(P<0.05).Bifidobacteriaceae showed high abundance in group X(P<0.001).Verrucomicrobiota showed low abundance in group M(P<0.05),and high abundance in group Y(P<0.05).Conclusion:Yantiao Decoction can not only regulate the level of inflammatory factors in sepsis rats,reduce excessive inflammatory response,but also improve the diversity and richness of intestinal microflora in sepsis rats.