亮菌口服液对5-FU化疗后小鼠肠黏膜屏障及肠道菌群移位的影响

Effect of armillariella oral solution on intestinal mucosal barrier and translocation of intestinal flora in mice after 5-FU chemotherapy

目的探讨亮菌口服液(AOS)对5-氟尿嘧啶(5-FU)化疗后小鼠肠黏膜屏障和肠道菌群移位的影响。方法将36只C57BL/6小鼠随机分为6组(n=6),正常组、模型组、亮菌口服液低、中、高剂量组(1、5、10 ml/kg)、双歧杆菌组(CBL,450 mg/kg)。腹腔注射5-FU 50 mg/kg 7 d建立小鼠肠黏膜损伤模型。HE染色检测小鼠肠黏膜损伤情况;通过异硫氰酸荧光素-葡聚糖(FITC-Dextran)示踪法检测肠黏膜通透性;酶联免疫吸附试验(ELISA法)检测血浆中内毒素含量;细菌平板计数法检测肠道细菌移位情况;免疫组化检测肠道紧密连接蛋白带状闭合蛋白(ZO-1)、咬合蛋白(occludin)的表达情况。结果①与正常组比较,模型组HE染色提示小鼠肠黏膜损伤明显,绒毛萎缩融合,隐窝加深或消失,大量炎性细胞浸润;血浆中FITC-Dextran及内毒素含量明显升高(P<0.05),肠道细菌移位率显著升高(P<0.01);免疫组化提示ZO-1、occludin的表达明显下降(P<0.01);②与模型组比较,AOS中、高剂量组小鼠肠黏膜损伤减轻,绒毛和隐窝部分恢复,少量炎性细胞浸润;血浆中FITC-Dextran及内毒素含量明显降低(P<0.05),肠道细菌移位率明显降低(P<0.05);ZO-1、occludin蛋白表达明显升高(P<0.01)。结论 AOS能缓解5-FU化疗后小鼠的肠黏膜损伤,其机制可能与降低肠黏膜的通透性,抑制内毒素和细菌移位有关。

Objective To investigate the effects of armillariella oral solution(AOS) on intestinal mucosal barrier and intestinal flora translocation after fluorouracil(5-FU) chemotherapy.Methods A total of 36 C57 BL/6 mice were randomly divided into 6 groups(n=6), the normal group, the model group, the low, medium and high dose groups of AOS(1,5,10 ml/kg), combined Bifidobacterium and Lactobacillus tablets group(CBL 450 mg/kg). A mouse intestinal mucosal injury model was established by intraperitoneal injection of 5-FU(50 mg/kg) for 7 days.HE staining was used to detect intestinal mucosal injury in mice. Intestinal mucosal permeability was detected by fluorescein-dextran( FITC-Dextran) tracer method. Endotoxin content in plasma was detected by enzyme linked immunosorbent assay( ELISA). Intestinal bacterial translocation was detected by bacterial plate count method. The expression of tight junction protein ZO-1 and occludin in small intestine was detected by immunohistochemistry( IHC). Results Compared with the normal group,HE staining showed that the intestinal mucosal injury of the model group was obvious that the villus was atrophied and merged,the crypt was deepened or disappeared,and a large number of inflammatory cells were infiltrated;The content of FITC-Dextran and endotoxin in plasma was significantly increased( P < 0. 05) as well as the intestinal bacterial translocation rate( P < 0. 01);Immunohistochemistry showed that the expression of ZO-1 and occludin were decreased significantly( P < 0. 01);Compared with the model group,intestinal mucosal damage was alleviated that villus and crypt were partially restored,and less inflammatory cells were infiltrated;The levels of FITC-Dextran and endotoxin in plasma were significantly decreased( P <0. 05) as well as intestinal bacterial translocation rate( P < 0. 05);While the expression of ZO-1,occludin was significantly increased( P < 0. 01) in the medium and high dose groups of AOS. Conclusion AOS may alleviate intestinal mucosal injury after 5-FU chemotherapy which may be related to reducing intestinal mucosal permeability and inhibiting endotoxin and bacterial translocation.