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

田瑞琦; 张诗捷; 丁银川; 薛敏

doi:10.13748/j.cnki.issn1007-7693.20240529

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

摘要

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小鼠随机分为对照组、顺铂组、低剂量绿原酸组及高剂量绿原酸组，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HE染色及组织病理观察并通过Western blotting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。
结果与对照组比较，顺铂组小鼠结肠组织中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)；绿原酸高剂量干预可明显降低三种炎性因子mRNA水平(P<0.01)，增加Occludin蛋白表达(P<0.01)，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对照组占优势；而相较于对照组，厚壁菌门和疣微菌门在顺铂组中优势明显增加(P<0.01)；此外相较于对照组，厚壁菌门和拟杆菌门比值在顺铂组升高(P<0.01)，而使用绿原酸低剂量和高剂量均可降低其比值(P<0.05或P<0.01)；在属水平上，相比对照组，顺铂组拟杆菌属、乳酸杆菌属、阿克曼菌属和毛罗菌科_NK4A136菌属相对丰度减少(P<0.05或P<0.01)，柠檬酸杆菌属相对丰度增加(P<0.01)；而绿原酸低剂量干预可部分恢复拟杆菌属及乳酸杆菌属相对丰度(P<0.05或P<0.01)，抑制柠檬酸杆菌属相对丰度(P<0.01)，高剂量组干预可增加阿克曼菌属及毛罗菌科_NK4A136菌属相对丰度(P<0.01)。
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 group, cisplatin group, low dose of chlorogenic acid group and high dose of chlorogenic acid 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 blotting and histopathological observation was carried by HE staining. The composition of intestinal flora in cecum contents was determined by 16s rDNA high-throughput sequencing.
RESULTS Compared with control group, the mRNA levels of IL-1β, IL-6 and TNF-α in colon tissue of cisplatin group were significantly increased(P<0.01), and the expression of Occludin protein in colon tissue was decreased(P<0.01). Chlorogenic acid intervention could significantly reduce the mRNA level of inflammatory factors(P<0.01), increase the expression of Occludin protein(P<0.01), and improve the damage of colonic mucosal barrier and inflammatory injury induced by cisplatin. 16s rDNA high-throughput sequencing results showed that Bacteroidota and firmicutes were dominant in the control group at the phylum level. Compared with the control group, Firmicutes and Verrucomicrobia were significantly more dominant in cisplatin group(P<0.01). In addition, compared with the control group, the ratio of Firmicutes to Bacteroidota was increased in cisplatin group(P<0.01), while the ratio was decreased by low and high doses of chlorogenic acid(P<0.05 or P<0.01). At the genus level, compared with the control group, the relative abundance of Bacteroides, Cillobacterium, Ackermania and Lachnospiraceae_NK4A136 decreased(P<0.05 or P<0.01), and the relative abundance of Citrobacter increased(P<0.01) in cisplatin group. The low dose of chlorogenic acid could partially restore the relative abundances of Bacteroides and Cillobacterium(P<0.05 or P<0.01), inhibit the relative abundances of Citrobacter(P<0.01), and increase the relative abundances of Akermania and Lachnospiraceae_NK4A136(P<0.01).
CONCLUSION Chlorogenic acid can improve intestinal mucosal barrier destruction induced by cisplatin, reduce intestinal inflammation, and partially restore intestinal flora disturbance caused by cisplatin.

HTML全文

参考文献(20)

施引文献

资源附件(0)