基于肠道菌群变化探讨健脾泄浊解毒方预防缓解期溃疡性结肠炎大鼠复发的实验研究

Preventing recurrence of Jianpi Xiezhuo Jiedu Recipe on inactive ulcerative colitis rats:based on intestinal flora

目的 基于肠道菌群变化探讨健脾泄浊解毒方对缓解期溃疡性结肠炎(UC)模型大鼠复发的干预效果。方法 在85只大鼠中随机取10只作为空白组,剩余75只大鼠通过结肠灌注2,4,6-三硝基苯磺酸(TNBS)与50%乙醇的混合液进行第1次造模(期间死亡3只),再将剩余72只大鼠随机分为模型组、柳氮磺胺吡啶组、健脾泄浊解毒方低剂量组及健脾泄浊解毒方高剂量组,每组18只,各组再随机处死2只大鼠观察以确定造模成功。除空白组正常喂养外,模型组予0.9%氯化钠注射液10 mL/kg灌胃,柳氮磺胺吡啶组予柳氮磺胺吡啶混悬液0.3 g/kg灌胃,健脾泄浊解毒方低、高剂量组分别予健脾泄浊解毒方药液8.0、16.0 g/kg灌胃。灌胃14 d后空白组随机取5只大鼠,其余各组随机取8只大鼠,处死后留取粪便标本,并将模型组、柳氮磺胺吡啶组、健脾泄浊解毒方低剂量组及健脾泄浊解毒方高剂量组剩余大鼠再次予TNBS与50%乙醇的混合液进行第2次灌注造模,继续正常喂养2 d后,各组大鼠全部处死,留取粪便标本。比较各组大鼠第1次造模治疗后及第2次造模后粪便标本中乳酸杆菌、双歧杆菌、肠杆菌及肠球菌DNA相对表达情况。结果 与第1次造模治疗后比较,柳氮磺胺吡啶组及健脾泄浊解毒方低剂量组第2次造模后乳酸杆菌及双歧杆菌相对表达量均降低(P<0.05),肠杆菌及肠球菌相对表达量均升高(P<0.05),空白组、模型组及健脾泄浊解毒方高剂量组第2次造模后乳酸杆菌、双歧杆菌、肠杆菌及肠球菌相对表达量比较差异无统计学意义(P>0.05)。第2次造模后组间比较,模型组乳酸杆菌及双歧杆菌相对表达量均低于空白组(P<0.05),肠杆菌及肠球菌相对表达量均高于空白组(P<0.05);健脾泄浊解毒方高剂量组乳酸杆菌及双歧杆菌相对表达量均高于模型组(P<0.05),肠杆菌及肠球菌相对表达量均低于模型组(P<0.05),而柳氮磺胺吡啶组及健脾泄浊解毒方低剂量组乳酸杆菌、双歧杆菌、肠杆菌及肠球菌相对表达量与模型组比较差异均无统计学意义(P>0.05)。结论 高剂量健脾泄浊解毒方能以肠道菌群为作用靶点,通过提高乳酸杆菌、双歧杆菌相对含量,降低肠杆菌、肠球菌相对含量,使有益菌较有害菌数量占据主导,调节肠道菌群紊乱状态,进而维持肠道微环境的长久稳态,保持机体健康,有效预防UC复发。

Objective To explore the effect of Jianpi Xiezhuo Jiedu Recipe(JPXZJDR)in preventing recurrence of inactive ulcerative colitis in rats based on intestinal flora.Methods Of the 85 rats,10 rats were randomized as Blank group(normal feeding),and the remaining 75 rats participated in the first modeling by intracolonic administration of 2,4,6-trinitrobenzenesulfonic acid(TNBS)dissolved in 50%ethanol.After 3 days of feeding,the obtained 72 rats were randomly grouped at 1∶1∶1∶1 ratio,and 2 rats from each group were randomly sacrificed to determine whether the first modeling was successful.The remaining rats were gavaged:Model group(0.9%sodium chloride solution 10 mL·kg^(-1)·d^(-1)),Salazosulfapyridine group(SASP 0.3 g·kg^(-1)·d^(-1)),low-,high-dose JPXZJDF group(JPXZJDF 8.0 g·kg^(-1)·d^(-1),16.0 g·kg^(-1)·d^(-1)).After 14 days of treatment,5 rats from Blank group and 8 rats from the other groups were randomly sacrificed for collection of fecal samples.Then,the remaining rats in Model group,SASP group and JPXZJDF groups participated in the second modeling by intracolonic administration of same TNBS as the first modeling.After 2 days of normal feeding,the all rats in Blank group and Modeling groups were sacrificed to collect stool samples again.Result SASP group and low-dose JPXZJDF group had significant lower relative content of Lactobacillus and Bifidobacterium,and significant higher relative content of Enterobacter and Enterococcusat at the second modeling vs the first modeling(all P<0.05);No significant difference was observed in the above indexes between Blank group and Model group,between Blank group and high-dose JPXZJDF group,and between Model group and high-dose JPXZJDF group(all P>0.05).In comparison between groups after the second modeling,Model group had significant lower relative content of Lactobacillus and Bifidobacterium,and significant higher relative content of Enterobacter and Enterococcus than Blank group(all P<0.05);high-dose JPXZJDF group had significant higher of Lactobacillus and Bifidobacterium,and significant lower relative content of Enterobacter and Enterococcus than Model group(all P<0.05);there were no differences in the above indexes at the second modeling between SASP group and low-dose JPXZJDF group,between SASP group and Model group,between low-dose JPXZJDF group and model group(all P>0.05).Conclusion Based on intestinal flora,High-dose JPXZJDF can act on UC targets.By elevated the relative content of Lactobacillus and Bifidobacterium,as well as reduced the relative content of Enterobacter and Enterococcus,which make beneficial bacteria play a dominant role.The aiming is to contribute to the regulation intestinal flora disturbance,maintain the long-term stability of the intestinal microenvironment,and effectively prevent UC recurrence.