[关键词]
[摘要]
目的 采用 16S rRNA 高通量测序技术探讨火麻仁油对非酒精性脂肪性肝炎(NASH)小鼠肠道菌群的影 响。方法 将 18 只 C57BL/6J 雄性小鼠随机分为正常组、模型组和火麻仁油组(1 g·kg-1 ),每组 6 只。正常组 喂养蛋氨酸-胆碱充足(MCS)饲料,其余两组喂养蛋氨酸-胆碱缺乏(MCD)饲料 4 周复制 NASH 小鼠模型。模 型复制期间火麻仁油组灌胃 1 g·kg-1火麻仁油,其余两组给予等体积 0.5% 吐温-80。实验过程中记录各组小鼠 体质量,4 周后收集血清、肝脏和粪便样本。采用苏木精-伊红(HE)、Masson 和油红 O 染色法观察肝脏组织病 理损伤;采用酶联免疫吸附测定法(ELISA)测定血清中丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶 (AST)的含量;采用 16S rRNA 高通量测序技术检测小鼠肠道菌群的变化。结果 与正常组比较,模型组小鼠 的体质量明显降低(P<0.01),血清 ALT 和 AST 的含量明显升高(P<0.001),肝脏内出现大量脂滴堆积、脂肪 变性,伴有大量炎性细胞浸润,且肝脏发生纤维化。与模型组比较,火麻仁油能够明显改善 NASH 小鼠的体 质量减轻(P<0.05),明显降低血清中 ALT 和 AST 的水平(P<0.01),并能显著改善肝脏病理损伤。16S rRNA 测序结果显示,火麻仁油可以逆转 NASH 小鼠肠道菌群 Alpha 与 Beta 多样性的下降。此外,鉴定了 19 种与 NASH 相关的菌属,火麻仁油可以对其中 7 种产生有效的调节。结论 火麻仁油可以改善 NASH 小鼠肝组织的 脂滴堆积、脂质变性、炎症反应和纤维化,其作用机制可能是通过调节肠道菌群多样性,促进有益菌和抑制有 害菌的生长来改善 NASH。
[Key word]
[Abstract]
Objective The effect of hemp seed oil on intestinal flora in non-alcoholic steatohepatitis (NASH) mice was investigated by 16S rRNA high-throughput sequencing technology. Methods Eighteen C57BL/6J male mice were randomly divided into normal group,model group and hemp seed oil group (1 g·kg-1 ),with 6 mice in each group. The normal group was fed with methionine-choline sufficiency(MCS) diet, and the other two groups were fed with methionine-choline deficient (MCD) diet for 4 weeks to replicate NASH mouse model. During the model replication, the hemp seed oil group was given 1 g·kg-1 hemp seed oil,and the other two groups were given equal volume of 0.5% Tween-80. The body mass of mice in each group was recorded during the experiment. After 4 weeks,serum,liver and fecal samples were collected. The pathological damage of liver tissue was detected by hematoxylin-eosin (HE),Masson and oil red O staining. The contents of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were determined by enzyme-linked immunosorbent assay (ELISA). The changes of gut microbiota in mice were detected by 16 S rRNA high-throughput sequencing technology. Results Compared with the normal group,the body mass of the mice in the model group was significantly reduced(P<0.01), and the contents of serum ALT and AST were significantly increased(P<0.001). A large number of lipid droplets and steatosis were observed in the liver, accompanied by a large number of inflammatory cell infiltration and liver fibrosis. Compared with the model group, hemp seed oil could significantly improve the body mass loss of NASH mice (P<0.05),significantly reduce the levels of ALT and AST in serum (P<0.01),and significantly improve the pathological damage of liver. The results of 16S rRNA sequencing showed that hemp seed oil could reverse the decrease of Alpha and Beta diversity of gut microbiota in NASH mice. In addition,19 NASH-related genera were identified,and hemp seed oil could effectively regulate 7 of them. Conclusion Hemp seed oil can improve lipid droplet accumulation,lipid degeneration,inflammatory response and fibrosis in liver tissue of NASH mice. The mechanism may be to improve NASH by regulating the diversity of gut microbiota,promoting the growth of beneficial bacteria and inhibiting the growth of harmful bacteria.
[中图分类号]
R285.5
[基金项目]
广东省普通高校特色创新类项目(2021KTSCX051)。