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基于肠道菌群和代谢组学研究金刚藤胶囊治疗非酒精性脂肪肝的作用及机制
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| 篇名： | 基于肠道菌群和代谢组学研究金刚藤胶囊治疗非酒精性脂肪肝的作用及机制 |
| TITLE： | Effect and mechanism of Jingangteng capsules in the treatment of non-alcoholic fatty liver disease based on gut microbiota and metabolomics |
| 摘要： | 目的 探究金刚藤胶囊治疗非酒精性脂肪性肝病（NAFLD）的作用及机制。方法将32只SD大鼠随机分为正常组和造模组，造模组以高脂饲料喂养构建NAFLD大鼠模型。将造模成功的大鼠随机分为模型组、阿托伐他汀组[阳性对照，2mg/（kg·d）]和金刚藤胶囊低、高剂量组[0.63、2.52mg/（kg·d）]，每组6只。采用苏木素-伊红染色、油红O染色观察肝脏病理变化；酶联免疫吸附试验测定大鼠血清甘油三酯（TG）、总胆固醇（TC）、高密度脂蛋白胆固醇（HDL-C）、低密度脂蛋白胆固醇（LDL-C）、丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、肿瘤坏死因子α（TNF-α）、白细胞介素1β（IL-1β）、IL-6、IL-18水平。应用16SrDNA测序和非靶向代谢组学技术分析金刚藤胶囊对NAFLD大鼠肠道菌群和代谢的调控作用；基于代谢组学结果，采用Westernblot法对NAFLD大鼠肝脏中核因子κB（NF-κB）/NOD样受体家族蛋白3（NLRP3）信号通路相关蛋白进行检测。结果金刚藤胶囊可以显著降低NAFLD大鼠TG、TC、LDL-C、AST、ALT、TNF-α、IL-1β、IL-6、IL-18水平，提高HDL-C水平（P＜0.05或P＜0.01），减轻肝脏细胞脂肪变性及炎性浸润；能调节NAFLD大鼠的肠道菌群紊乱，显著增加Oscillospira、Romboutsia两种菌属的相对丰度，显著降低Blau‐tia的相对丰度（P＜0.05）；主要通过影响次级胆汁酸生物合成、脂肪酸降解、O-抗原核苷酸糖生物合成等途径调节代谢紊乱。Westernblot法检测结果显示，该药可以显著降低NF-κBp65、NF-κB抑制因子α的磷酸化水平和NLRP3、胱天蛋白酶1、抗体分泌细胞蛋白表达水平（P＜0.05或P＜0.01）。结论金刚藤胶囊可改善NAFLD大鼠的炎症反应、脂质积累和肝脏损伤，调节肠道微生态及代谢紊乱，通过抑制NF-κB/NLRP3信号通路发挥治疗NAFLD的作用。 |
| ABSTRACT： | OBJECTIVE To investigate the effect and mechanism of Jingangteng capsules in the treatment of non-alcoholic fatty liver disease （NAFLD）. METHODS Thirty-two SD rats were randomly divided into normal group and modeling group. The modeling group was fed a high-fat diet to establish a NAFLD model. The successfully modeled rats were then randomly divided into model group， atorvastatin group[positive control， 2 mg/（kg·d）]， and Jingangteng capsules low- and high-dose groups [0.63 and 2.52 mg/（kg·d）]， with 6 rats in each group. The pathological changes of the liver were observed by hematoxylin-eosin staining and oil red O staining. Enzyme-linked immunosorbent assay was performed to determine the serum levels of triglycerides （TG）， total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， alanine transaminase （ALT）， aspartate transaminase （AST）， tumour necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， IL-6， IL-18. 16S rDNA amplicon sequencing and metabolomics techniques were applied to explore the effects of Jingangteng capsules on gut microbiota and metabolisms in NAFLD rats. Based on the E-mail：591146765@qq.com metabolomics results， Western blot analysis was performed to detect proteins related to the nuclear factor kappa-B （NF-κB）/NOD-like receptor family protein 3 （NLRP3） signaling pathway in the livers of NAFLD rats. RESULTS The experimental results showed that Jingangteng capsules could significantly reduce the serum levels of TG， TC， LDL-C， AST， ALT， TNF-α， IL-1β， IL-6， IL-18， while increased the level of HDL-C， and alleviated the hepatic cellular steatosis and inflammatory infiltration in NAFLD rats. They could regulate the gut microbiota disorders in NAFLD rats， significantly increased the relative abundance of Romboutsia and Oscillospira， and significantly decreased the relative abundance of Blautia （P＜0.05）. They also regulated metabolic disorders primarily by affecting secondary bile acid biosynthesis， fatty acid degradation， O-antigen nucleotide sugar biosynthesis， etc. Results of Western blot assay showed that they significantly reduced the phosphorylation levels of NF-κB p65 and NF-κB inhibitor α， and the protein expression levels of NLRP3， caspase-1 and ASC （P＜0.05 or P＜0.01）. CONCLUSIONS Jingangteng capsules could improve inflammation， lipid accumulation and liver injury in NAFLD rats， regulate the disorders of gut microbiota and metabolisms， and inhibit NF-κB/NLRP3 signaling pathway. Their therapeutic effects against NAFLD are mediated through the inhibition of the NF-κB/NLRP3 signaling pathway. |
| 期刊： | 2025年第36卷第11期 |
| 作者： | 程世源；熊悦；张丹丹；李晶；孙志滢；田家瑛；沈丽；沈月；刘丹；魏琼；叶晓川 |
| AUTHORS： | CHENG Shiyuan，XIONG Yue， ZHANG Dandan，LI Jing，SUN Zhiying，TIAN Jiaying，SHEN Li，SHEN Yue，LIU Dan，WEI Qiong，YE Xiaochuan |
| 关键字： | 金刚藤胶囊；非酒精性脂肪肝；肠道菌群；代谢组学；NF-κB/NLRP3信号通路 |
| KEYWORDS： | Jingangteng capsules； non-alcoholic fatty liver disease； gut microbiota； metabolomics； NF-κB/NLRP3 signaling |
| 阅读数： | 5 次 |
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