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OBJECTIVES@#To investigate the mechanism of comorbidity between non-alcoholic fatty liver disease (NAFLD) and atherosclerosis (AS) based on metabolomics and network pharmacology.@*METHODS@#Six ApoE－/－ mice were fed with a high-fat diet for 16 weeks as a comorbid model of NAFLD and AS (model group). Normal diet was given to 6 wildtype C57BL/6J mice (control group). Serum samples were taken from both groups for a non-targeted metabolomics assay to identify differential metabolites. Network pharmacology was applied to explore the possible mechanistic effects of differential metabolites on AS and NAFLD. An in vitro comorbid cell model was constructed using NCTC1469 cells and RAW264.7 macrophage. Cellular lipid accumulation, cell viability, morphology and function of mitochondria were detected with oil red O staining, CCK-8 assay, transmission electron microscopy and JC-1 staining, respectively.@*RESULTS@#A total of 85 differential metabolites associated with comorbidity of NAFLD and AS were identified. The top 20 differential metabolites were subjected to network pharmacology analysis, which showed that the core targets of differential metabolites related to AS and NAFLD were STAT3, EGFR, MAPK14, PPARG, NFKB1, PTGS2, ESR1, PPARA, PTPN1 and SCD. The Kyoto Encyclopedia of Genes and Genomes showed the top 10 signaling pathways were PPAR signaling pathway, AGE-RAGE signaling pathway in diabetic complications, alcoholic liver disease, prolactin signaling pathway, insulin resistance, TNF signaling pathway, hepatitis B, the relax in signaling pathway, IL-17 signaling pathway and NAFLD. Experimental validation showed that lipid metabolism-related genes PPARG, PPARA, PTPN1, and SCD were significantly changed in hepatocyte models, and steatotic hepatocytes affected the expression of macrophage inflammation-related genes STAT3, NFKB1 and PTGS2; steatotic hepatocytes promoted the formation of foam cells and exacerbated the accumulation of lipids in foam cells; the disrupted morphology, impaired function, and increased reactive oxygen species production were observed in steatotic hepatocyte mitochondria, while the formation of foam cells aggravated mitochondrial damage.@*CONCLUSIONS@#Abnormal lipid metabolism and inflammatory response are distinctive features of comorbid AS and NAFLD. Hepatocyte steatosis causes mitochondrial damage, which leads to mitochondrial dysfunction, increased reactive oxygen species and activation of macrophage inflammatory response, resulting in the acceleration of AS development.
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Animals , Mice , Non-alcoholic Fatty Liver Disease/metabolism , Cyclooxygenase 2/metabolism , PPAR gamma/metabolism , Reactive Oxygen Species/metabolism , Mice, Inbred C57BL , Hepatocytes , Macrophages/metabolism , LiverABSTRACT
BACKGROUND: Naoxinqing capsule has been used for treating cerebral ischemia/reperfusion injury for a long time. However, there are relatively few in-depth studies on its mechanism. OBJECTIVE: To investigate the therapeutic effect of Naoxinqing Capsule on gerbil model of cerebral ischemia/reperfusion injury by molecular biological means. METHODS: The study was approved by the Laboratory Animal Ethical Committee of Liaoning University of Chinese Medicine, approval No. 21000092017072. Eighty male Mongolian gerbils were randomly divided into sham, model, Naoxinqing and Naoluotong groups, and the latter three groups underwent bilateral common carotid artery clip for 5 minutes, to establish the model of cerebral ischemia/reperfusion injury. The sham group received no common carotid artery clip. Next day, the sham group fed normally, the model group was given normal saline, Naoxinqing group was given the 100 mg/(kg•d) Naoxinqing via gavage, and Naoluotong group given 100 mg/(kg•d) Naoluotong via gavage, respectively, for 21 consecutive days. The water maze test was conducted at 1 week before experiment ended. The brain tissue was removed after experiment. The learning and memory function, hippocampal neurons, cerebrovascular and corresponding molecular changes were detected. RESULTS AND CONCLUSION: (1) Compared with the sham group, the learning ability in the model group was decreased significantly. Naoxinqing and Naoluotong groups could effectively improve the learning ability after surgery. (2) Compared with the model group, the numbers of neurons in the Naoxinqing and Naoluotong groups were increased significantly, arranged regularly with clear contour and complete structure. (3) Compared with the model group, in the Naoxinqing and Naoluotong groups, the activities of superoxide dismutase and lactate dehydrogenase, and glutathione content were significantly increased, and the content of malonaldehyde was significantly decreased (P < 0.01). (4) The expression levels of ASC, NLRP3 and Caspase-1 in the hippocampus in the Naoxinqing and Naoluotong groups were significantly lower than those in the model group (P < 0.05). (5) The levels of interleukin-18 and interleukin-1β in the Naoxinqing and Naoluotong groups were significantly lower than those in the model group (P < 0.01). (6) Compared with the model group, the cells positive for platelet endothelial cell adhesion molecule-1 in the Naoxinqing and Naoluotong groups were significantly increased, the cells contacted closely each other. (7) Compared with the model group, in the Naoxinqing and Naoluotong groups, the expression levels of platelet endothelial cell adhesion molecule-1 and phosphorylated endothelial nitric oxide synthase were significantly up-regulated, and the content of nitric oxide was significantly increased (P < 0.01). (8) These results indicate that Naoxinqing and Naoluotong can effectively protect the morphology of hippocampal CA1 region in gerbils. Cerebral ischemia/reperfusion injury is accompanied by cerebral vascular dysfunction. Naoxinqing Capsule can protect cerebral vascular function and inhibit cerebral ischemia/reperfusion injury.
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Objective To investigate the effect of a decoction to nourish qi and invigorate the spleen on mitochondrial respiratory chain enzyme complex activity in cardiomyocytes of rats with spleen qi deficiency syndrome. Methods Rats were randomly divided into normal,model,and treatment groups. The model and treatment groups were treated by diet intervention combined with the limit swim method. The general condition and spleen qi deficiency syndrome were assessed on day 15. After the success of the model,the normal and model groups were treated with a con?ventional feeding method combined with normal saline ,and the treatment group was treated by diet intervention combined with a decoction to nour?ish qi and invigorate the spleen for 9 weeks. The activity of two mitochondrial respiratory chain enzyme complexes was observed. Results The ac?tivity of mitochondrial respiratory chain enzyme complexⅡand complexⅣin the model group was significantly lower than the activity in the nor?mal and treatment groups(P<0.05). The activity levels of complexⅡand complexⅣwere significantly different between the model group and the treatment group(P<0.05). Conclusion Spleen qi deficiency can cause decreased activity of mitochondrial respiratory chain enzyme com?plexes in myocardial cells. The decoction to nourish qi and invigorate the spleen can modulate the activity of myocardial mitochondrial respiratory chain enzyme complexesⅡandⅣ.
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<p><b>OBJECTIVE</b>To explore the effects of astragalus (AST) , total flavone of astragalus (TFA), total saponins of astragalus (TSA) and astragalus polysaccharides (APS) on ischemia/reperfusion (40 min/60 min) injury in isolated guinea-pig heart.</p><p><b>METHODS</b>Isolated guinea-pig hearts underwent ischemia, then followed by K-H perfusion (I/R group), AST (60 mg/L),AST (60 mg/L), TFA (60 mg/L), TSA (60 mg/L) and APS (60 mg/L) perfusion (n = 6 each).Isolated hearts without ischemia serve as control group (n = 6). Activity of lactate dehydrogenas (LDH) and creatine kinase (CK) in effluent were measured.Infarct size, myocardial superoxide dismutase (SOD) activity and malondiadehyde (MDA) contents were also determined.</p><p><b>RESULTS</b>Compared to control hearts, heart rate, coronary flow and myocardial superoxide dismutase (SOD) activity were significantly reduced, while LDH and CK in effluent as well as myocardial MDA were significantly increased in the I/R hearts during reperfusion (all P < 0.05), these changes could be partly reversed by AST and TFA perfusion.Infarct size was also significantly reduced in AST (11.9 ± 2.03) % and TFA (13.31 ± 1.17) % treated hearts compared to that in I/R group (18.9 ± 2.27) % (all P < 0.01).</p><p><b>CONCLUSIONS</b>The findings indicate that AST and TFA could attenuate I/R injury in isolated guinea-pig heart possibly through enhancing the activity of SOD and reducing lipid peroxidation.</p>