Exploring the Mechanism of Jiangzhi Qingshen Capsules Mediating NF-κB/NLRP3/Caspase-1 Signaling Axis to Improve Hyperlipidemia in Mice Based on Transcriptomics and Weighted Gene Co-expression Network Analysis
Objective Based on transcriptomics and weighted gene co-expression network analysis (WGCNA),this study aims to explore the mechanism of Jiangzhi Qingshen Capsules mediating the nuclear transcription factor(NF)-κB/NOD like receptor heat protein domain related protein 3(NLRP3)/cysteine protease-1(Caspase-1)signaling axis to improve hyperlipidemia. Method Forty C57BL/6 mice were randomly divided into control group,model group,rosuvastatin group (1.3 mg·kg-1),Jiangzhi Qingshen Capsules low-and high-dose groups (0.6 and 2.4 g·kg-1),with eight mice in each group. Except the control group was given regular feed,other groups were given high-fat feed to replicate the hyperlipidemia model. The treatment groups were orally administered the corresponding drug once a day for 10 consecutive weeks. The mass of rat liver was taken to calculate the liver index,Lee's index and ratio of liver weight to tibia length. Hematoxylin eosin (HE)staining and oil red O staining were used to observe pathological changes in liver tissue. Serum biochemical indicators including total cholesterol (TC),triglycerides (TG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C),alanine aminotransferase(ALT),aspartate aminotransferase (AST),interleukin (IL)-18 and IL-1β were detected by ELISA. The transcriptome sequencing of liver tissue was performed using Bo'ao Jingdian mouse eukaryotic transcriptome chip V4,then WGCNA of chip expression information and the trait data of blood lipid levels was carried out. The core gene groups (potential targets) of Jiangzhi Qingshen Capsule intervention in hyperlipidemia were obtained,and they were further used to perform protein-protein interaction (PPI) network analysis,Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analysis. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect the mRNA levels of NF-κB,NLRP3,Caspase-1,IL-18,and IL-1β in liver,and immunofluorescence was used to detect the protein expression of NF-κB,NLRP3 and Caspase-1 in liver. Results Compared with model group,Jiangzhi Qingshen Capsules effectively reduced the body weight,liver wet weight,liver index,Lee's index and ratio of liver weight to tibia length (P<0.05),effectively alleviate inflammatory infiltration and steatosis in liver tissue of hyperlipidemic mice,and significantly reduce lipid deposition area (P<0.05). Moreover,Jiangzhi Qingshen Capsules obviously improved serum TC,LDL-C,ALT,AST,IL-18,and IL-1β levels(P<0.05),effectively down-regulated the protein and mRNA expression of NF-κB,NLRP3 and Caspase-1 in liver tissue (P<0.05),and decreased the mRNA levels of IL-18 and IL-1β (P<0.05). Transcriptomics and WGCNA showed that the regulatory effect of Jiangzhi Qingshen Capsules on blood lipids may be closely related to the inflammatory pathway. Conclusion Jiangzhi Qingshen Capsules can inhibit inflammatory reaction in liver tissue of hyperlipidemic mice by mediating the NF-κB/NLRP3/Caspase-1 signaling axis,thereby improving blood lipid levels.
万方数据
维普
