| Research ObjectiveTo explore the beneficial effects and related mechanisms of Rhizoma Gastrodiae(Rg)in regulating the gut microbiota on ligature-induced carotid artery injury in apolipoprotein E gene knockout(ApoE-/-)mice by combining 16S rRNA gene sequencing analysis and untargeted metabolomics techniques,providing scientific evidence for the treatment of atherosclerosis(AS)with Rg.Research Content(1)Observational study of the clinical efficacy of Rhizoma Gastrodiae capsule treatment on Carotid Atherosclerosis(CAS):the treatment group(n=30)under Gastrodia intervention was compared with the control group(n=30)regarding gender,age,smoking history,drinking history,BMI index,blood pressure;hypertension,type-2 diabetes,hyperlipidemia;routine blood tests,liver and kidney function,lipid levels,electrolytes;abdominal ultrasound,carotid artery ultrasonography changes to evaluate efficacy.(2)The treatment effect of Rg on intimal hyperplasia in AS mice:16 male ApoE-/-mice underwent carotid artery ligation and were randomized into control and treatment groups,8 mice each.The treatment group received an oral dose of 10mg/kg/d Gastrodia,and the control group received an equivalent volume of saline.A high-fat diet was provided for 12 weeks to establish the atherosclerosis(AS)model,followed by sampling.Vascular tissue morphology was observed through Oil Red O staining.Western Blot(WB)detection of Cyclin D1 and CDK4 expression levels determined the extent of intimal hyperplasia.Proliferation and apoptosis in carotid artery tissues were observed through immunofluorescence staining.(3)Exploring the mechanism by which Rg ameliorates ligature-induced intimal hyperplasia in AS mice based on gut microbiota:Building on the previous study,using 16S rRNA gene sequencing analysis to investigate the impact of Rg on the alpha and beta diversity of the gut microbiota of AS mice with intimal hyperplasia.(4)Investigating the impact of Rg on the serum metabolic profiles of AS mice with intimal hyperplasia using untargeted metabolomics technology:Mouse serum was collected for untargeted metabolomics analysis to screen differential metabolic products.Differential metabolites were subjected to enrichment analysis of metabolic pathways using MetaboAnalyst 4.0,with KEGG as the database,to analyze the metabolomics of differential metabolites.Research Results(1)Rg improved carotid intima-media thickness,lipid levels,and fasting blood glucose control in patients with CAS,increased the overall efficiency of statin therapy for atherosclerotic plaques,while controlling patient weight and reducing the AIP index.(2)Rg reduced plaque area in ApoE-/-mice and increased the lumen area in ligated carotid artery tissue.Additionally,Rg decreased the content of vascular smooth muscle cells(VSMCs)in ligated carotid artery tissues by reducing VSMCs proliferation and increasing VSMCs apoptosis.(3)Rg intervention affected the alpha and beta diversity of gut microbiota in AS model mice with intimal hyperplasia,regulating the relative abundance at the phylum level of Firmicutes and Bacteroidetes,and at the genus level of Lactobacillus and Oscillospira.Concurrently,Rg caused changes in bacterial abundance at the phylum and genus levels of gut microbiota.(4)Rg affected the levels of metabolic products such as Albuterol,2,5-dihydroxybenzenesulfonate,all-trans-Retinoic acid,D-Glucuronic acid,6-Hydroxyflavone,Cortisol,etc.These metabolic products are related to Ascorbate and aldarate metabolism,Histidine metabolism,and Retinol metabolism pathways.Research ConclusionThis study demonstrated the beneficial effects and potential mechanisms of Rg in an ApoE-/-mouse model with ligature-induced carotid artery injury,emphasizing the potential of Rg as a therapeutic agent.By modulating the structure of the gut microbiome and intestinal metabolic pathways,Rg alleviated intimal hyperplasia by affecting VSMCs proliferation and apoptosis. |
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