香烟烟雾成分（CSCs）诱导大鼠血脂异常和肝脂肪变性的机制。
Induction mechanism of cigarette smoke components (CSCs) on dyslipidemia and hepatic steatosis in rats.
机构信息
College of Life Sciences, China Jiliang University, 258 XueYuan Street, XiaSha Higher Education Zone, Hangzhou, 310018, Zhejiang Province, People's Republic of China.
Institute of Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou, 310021, Zhejiang, China.
出版信息
Lipids Health Dis. 2022 Nov 8;21(1):117. doi: 10.1186/s12944-022-01725-8.
OBJECTIVE
The purpose of this study was to explore the effect of cigarette smoke component (CSC) exposure on serum lipid levels in rats and the underlying molecular mechanism.
METHODS
Male SPF-grade SD rats were randomly divided into a control group and a CSC exposure group, with the CSC group being exposed to CSC for 6 weeks. RT-PCR and Western blotting methods were used to detect lipid metabolism gene expression in rats, and 16S RNA gene sequencing was used to detect the gut microbiota in the rat cecum. Rat serum exosomes were prepared and identified, and the interaction of exosomal miR-291a-3p and miR-126a-5p with AMPK and CYP7A1 was detected by a dual luciferase reporter gene assay (DLRG).
RESULTS
Serum indicators, including cholesterol levels and trimethylamine oxide (TMAO) content, were significantly affected in the CSC exposure group compared with the control group (P < 0.05), and the expression levels of adenylate-activated protein kinase (AMPK), acetyl-coenzyme A carboxylase (ACC) and HMG-CoA reductase (HMG-CoAR) genes were significantly increased (P < 0.05) in the liver, while the expression level of cholesterol 7α-hydroxylase (CYP7A1) was markedly decreased (P < 0.01). 16S rRNA gene sequencing of the gut microbiota in the rat cecum showed that the abundance of Firmicutes in the CSC group increased significantly at the phylum level, while the abundances of Bacteroidota and Spirochaetota were reduced significantly (P < 0.01). The relative abundance of Romboutsia, Turicibacter, and Clostridium sensu stricto increased significantly (P < 0.01), and the relative abundance of Prevotella, Muribaculaceae_norank, Lachnospiraceae NK4A136 group, Roseburia, Treponema, and Ruminococcus significantly decreased (P < 0.01) at the genus level. In addition, the exosome miR-291a-3p and miR-126a-5p levels were markedly regulated by CSC exposure (P < 0.01). The interactions of miR-291a-3p and miR-126a-5p with AMPK and CYP7A1 mRNA were also validated by the DLRG method.
CONCLUSIONS
In summary, the rat dyslipidemia induced by CSC exposure may be related to the interference of gut microbiota structure and interaction of miRNAs from serum exosomes with target mRNAs, which further regulated AMPK-ACC/CYP7A1 signaling in rats.
目的
本研究旨在探讨香烟烟雾成分（CSC）暴露对大鼠血清脂质水平的影响及其潜在的分子机制。
方法
雄性 SPF 级 SD 大鼠随机分为对照组和 CSC 暴露组，CSC 组暴露于 CSC 6 周。采用 RT-PCR 和 Western blot 方法检测大鼠脂质代谢基因表达，16S RNA 基因测序检测大鼠盲肠肠道微生物群。制备大鼠血清外泌体并进行鉴定，通过双荧光素酶报告基因检测（DLRG）检测外泌体 miR-291a-3p 和 miR-126a-5p 与 AMPK 和 CYP7A1 的相互作用。
结果
与对照组相比，CSC 暴露组大鼠血清指标（包括胆固醇水平和三甲胺氧化物（TMAO）含量）显著受影响（P<0.05），肝组织中腺苷酸活化蛋白激酶（AMPK）、乙酰辅酶 A 羧化酶（ACC）和 HMG-CoA 还原酶（HMG-CoAR）基因表达水平显著升高（P<0.05），而胆固醇 7α-羟化酶（CYP7A1）基因表达水平显著降低（P<0.01）。大鼠盲肠肠道微生物群 16S rRNA 基因测序显示，CSC 组厚壁菌门丰度在门水平上显著增加，而拟杆菌门和螺旋体门丰度显著降低（P<0.01）。Romboutsia、Turicibacter 和 Clostridium sensu stricto 的相对丰度显著增加（P<0.01），而 Prevotella、Muribaculaceae_norank、Lachnospiraceae NK4A136 组、Roseburia、Treponema 和 Ruminococcus 的相对丰度显著降低（P<0.01）。此外，CSC 暴露显著调节外泌体 miR-291a-3p 和 miR-126a-5p 水平（P<0.01）。通过 DLRG 方法还验证了 miR-291a-3p 和 miR-126a-5p 与 AMPK 和 CYP7A1 mRNA 的相互作用。
结论
总之，CSC 暴露引起的大鼠血脂异常可能与肠道微生物群结构的干扰以及血清外泌体 miRNA 与靶 mRNA 的相互作用有关，进一步调节了大鼠 AMPK-ACC/CYP7A1 信号通路。
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