生、熟大黄对湿热泄泻大鼠炎性细胞因子、胃肠激素与肠道菌群的影响

Effects of Raw and Cooked Dahuang(Rhubarb)on Inflammatory Cytokines,Gastrointestinal Hormones and Intestinal Flora in Rats with Damp-Heat Diarrhea

目的探究大黄“通因通用”治疗泄泻的作用机制。方法采用高温+高湿+高糖高脂饮食+致病因子方式造模,分为生理组(不造模)、模型组(给予生理盐水),生大黄组(生药0.05 g·kg^(-1)),熟大黄组(生药0.05 g·kg^(-1))4组,每组早晚两次给药,共给药5 d。每日最后一次给药3 h后检测各组泻下指数,最后一次给药3 h后用ELISA法检测大鼠血清中白细胞介素-10(interleukin-10,IL-10)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)、白细胞介素-2(interleukin-2,IL-2)、大鼠血清和结肠胃动素(motilin,MTL)、大鼠血管活性肠肽(vasoactive intestinal peptide,VIP)、大鼠生长抑素(somatostatin,SS)、大鼠胃泌素(gastrin,Gas)和大鼠血清P物质(substance P,SP)含量,用16sDNA技术检测大鼠粪便中菌群结构。结果与模型组相比生、熟大黄均可显著降低大鼠泻下指数(P<0.01)。与模型组相比生、熟大黄均可显著降低大鼠体内TNF-α、IL-2、IL-6和IL-1β水平(P<0.01),可显著升高IL-10水平,其中生大黄组效果最佳。生大黄和熟大黄可以显著降低腹泻大鼠血清MTL、VIP、GAS和SP,提高血清SS水平,其中熟大黄组在降低血清GAS、升高SS优于生大黄。生大黄显著降低了结肠VIP,熟大黄组结肠胃肠激素和生理组相比无显著差异。生大黄可显著降低致病菌(如Escherichia-Shigella)和部分条件致病菌(Enterorhabdus和Bacteroides),但益生菌较生理组仍显著偏低。熟大黄有益于部分益生菌(如Ruminococcus、Adlercreutzia和Lachnospiraceae)的增殖,但对于致病菌的无显著抑制作用。整体上比较,生大黄组和生理组的菌群结构更加接近,熟大黄组和模型组的菌群结构更为接近。结论生、熟大黄均有治疗湿热泄泻作用,生大黄偏向于通过抗炎和调节肠道菌群紊乱两个环节促进湿热泄泻大鼠的转归,熟大黄偏向于调控胃肠激素水平发挥止泻作用。

Objective To explore the mechanism of Dahuang(rhubarb)treating diarrhea with purgative method in the treatment of diarrhea.Methods The model was made by high temperature+high humidity+high sugar and high fat diet+pathogenic factors.The model was divided into four groups:physiological group,model group(normal saline),raw Dahuang(rhubarb)group(crude drug 0.05 g·kg^(-1)),cooked Dahuang(rhubarb)group(crude drug 0.05 g·kg^(-1)).Each group was given drugs twice in the morning and evening for a total of 5 days.The diarrhea index of each group was detected 3 hours after the last administration every day,and the serum levels of interleukin-10(IL-10),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6)and interleukin-2(IL-2)in rats were detected by ELISA method 3 hours after the last administration.The contents of motilin(MTL),vasoactive intestinal peptide(VIP),somatostatin(SS),gastrin(Gas)and substance P(SP)in rat serum colonic were also detected by ELISA method 3 hours after the last administration.16sDNA technique was used to detect the flora structure in rat feces.Results Both raw and cooked Dahuang(rhubarb)could significantly reduce the diarrhea index of rats(P<0.01)compared with the model group.Both raw and cooked Dahuang(rhubarb)can significantly reduce the levels of TNF-α,IL-2,IL-6 and IL-1βin rats compared with the model group.The level of IL-10 can be significantly increased(P<0.01),and the effect of raw Dahuang(rhubarb)group was the best.Raw and cooked Dahuang(rhubarb)can significantly reduce serum levels of MTL,VIP,GAS and SP and increase serum SS level in diarrhea rats.Cooked Dahuang(rhubarb)group was better than raw Dahuang(rhubarb)in reducing serum GAS and increasing SS.Raw Dahuang(rhubarb)significantly reduced colon VIP,and there was no significant difference in colon gastrointestinal hormones between cooked Dahuang(rhubarb)group and physiological group.Dahuang(rhubarb)can significantly reduce pathogenic bacteria(such as Escherichia-Shigella)and some conditional pathogens(Enterhabdus and Bacteroides),but probiotics were still significantly lower than those in the physiological group.Cooked Dahuang(rhubarb)was beneficial to some probiotics,for example,Ruminococcus,Adlercreutzia and Lachnospiraceae had no significant inhibitory effect on the proliferation of pathogenic bacteria.On the whole,the flora structure of raw Dahuang(rhubarb)group and physiological group was more similar,and that of cooked Dahuang(rhubarb)group and model group was more similar.Conclusion:Both raw and cooked Dahuang(rhubarb)have the effect of treating damp-heat diarrhea,and raw Dahuang(rhubarb)tends to promote the prognosis of damp-heat diarrhea rats through anti-inflammatory and regulating intestinal flora disorder.Cooked Dahuang(rhubarb)tends to regulate the levels of gastrointestinal hormones to play an antidiarrheal role.