绿原酸对顺铂诱导小鼠肠屏障功能障碍和肠道菌群的影响

Effects of chlorogenic acid on intestinal barrier dysfunction and intestinal flora induced by cisplatin in mice

关键词:  绿原酸  顺铂  肠黏膜  肠道菌群.

Key words:  chlorogenic acid  cisplatin  intestinal mucosa  intestinal flora.

摘要:

目的 探究绿原酸对顺铂致小鼠肠屏障功能障碍和肠道菌群的影响。方法 28只6周龄C57BL/6J小鼠随机分为对照(CN)组，顺铂(CDDP)组，绿原酸低剂量干预(CDDP+CGA1)组及绿原酸高剂量干预(CDDP+CGA2)组，RT-qPCR分别检测各组小鼠结肠组织中IL-1β、IL-6及TNF-α mRNA水平；取结肠组织进行H＆E染色及组织病理观察并通过免疫印迹测定 Occludin蛋白表达；取各组小鼠新鲜粪便，16S rDNA高通量测序检测盲肠内容物中肠道菌群的组成。结果 与CN组比较，CDDP组小鼠结肠组织中IL-1β、IL-6及TNF-α mRNA明显升高(P<0.01)，且结肠组织中Occludin蛋白表达降低(P<0.01)；绿原酸干预可明显降低炎性因子mRNA水平，增加Occludin蛋白表达，改善顺铂致结肠黏膜屏障的破坏和炎性损伤。16S rDNA高通量测序检测结果显示：在门水平，拟杆菌门和厚壁菌门在CN组占优势，而相较于CN组，厚壁菌门、疣微菌门及变形菌门在CDDP组中优势明显增加；此外相较于CN组和绿原酸干预组，拟杆菌门和厚壁菌门比值在CDDP模型组下降(P< 0.05)；在属水平上，与CN组比较，CDDP模型组拟杆菌属、乳酸杆菌属、毛罗菌科_NK4A136菌属相对丰度减少(P<0.05)，柠檬酸杆菌属相对丰度增加(P<0.05)，而绿原酸干预可明显增加乳酸杆菌属、拟杆菌属及阿克曼菌属的相对丰度(P<0.05)。结论 绿原酸能改善顺铂致肠黏膜屏障的破坏，降低肠道炎症，部分恢复顺铂所致肠道菌群紊乱。

Abstract:

OBJECTIVE To investigate the effect of chlorogenic acid on intestinal barrier dysfunction and intestinal flora disorder induced by cisplatin in mice. METHODS Twenty-eight 6-week-old C57BL/6J mice were randomly divided into control (CN) group, cisplatin (CDDP) group, low dose of chlorogenic acid treatment (CDDP +CGA1) group and high dose of chlorogenic acid treatment (CDDP+CGA2) group. The levels of IL-1β, IL-6 and TNF-α mRNA in colon tissues of each group were detected by RT-qPCR. The expression of Occludin protein was determined by western blot and histopathological observation is carried by H&E staining; The composition of intestinal flora in cecum contents was determined by 16S rDNA high-throughput sequencing. RESULTS Compared with CDDP + CGA and CN groups, the levels of IL-1β, IL-6 and TNF-α mRNA in colon tissue in CDDP group increased; the expression of occludin protein in colon tissue decreased and histopathological observation showed that CGA could improve the damage of colon mucosal barrier and inflammatory injury induced by cisplatin. 16S rDNA high-throughput sequencing results showed that Bacteroidetes and Firmicutes were dominant in CN group. However, Firmicutes, Verrucomicrobia and Proteobacteria were dominant in CDDP group. Compared with CN group and chlorogenic acid treatment group, the ratio of Bacteroidetes to Firmicutes decreased in CDDP model group (P < 0.05). At the genus level, compared with CN group, the relative abundances of Bacteroides, Lactobacillus and Mauriaceae in CDDP model group decreased (P < 0.05), while the relative abundance of Citrobacter increased (P<0.05). However, Chlorogenic acid intervention significantly increased the relative abundance of Lactobacillus, Bacteroides and Ackermannia (P<0.05). CONCLUSION Chlorogenic acid improved intestinal mucosal barrier destruction induced by cisplatin, reduced intestinal inflammation, and partially restore intestinal flora disturbance caused by cisplatin.