Mechanism of Didang Decoction in prevention of anti-atherosclerosis and hyperlipidemia by HPLC-Q-TOF-MS/MS and network pharmacology based on theory of "nutrients return to heart and fat accumulates in channels" / 中国中药杂志

Atherosclerosis(AS) is caused by impaired lipid metabolism, which deposits lipids in the intima, causes vascular fibrosis and calcification, and then leads to stiffening of the vascular wall. Hyperlipidemia(HLP) is one of the key risk factors for AS. Based on the theory of "nutrients return to the heart and fat accumulates in the channels", it is believed that the excess fat returning to the heart in the vessels is the key pathogenic factor of AS. The accumulation of fat in the vessels over time and the blood stasis are the pathological mechanisms leading to the development of HLP and AS, and "turbid phlegm and fat" and "blood stasis" are the pathological products of the progression of HLP into AS. Didang Decoction(DDD) is a potent prescription effective in activating blood circulation, removing blood stasis, resolving turbidity, lowering lipids, and dredging blood vessels, with the functions of dispelling stasis to promote regeneration, which has certain effects in the treatment of atherosclerotic diseases. This study employed high-performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry(HPLC-Q-TOF-MS/MS) to screen the main blood components of DDD, explored the targets and mechanisms of DDD against AS and HLP with network pharmacology, and verified the network pharmacological results by in vitro experiments. A total of 231 blood components of DDD were obtained, including 157 compounds with a composite score >60. There were 903 predicted targets obtained from SwissTargetPrediction and 279 disease targets from GeneCards, OMIM, and DisGeNET, and 79 potential target genes of DDD against AS and HLP were obtained by intersection. Gene Ontology(GO) analysis suggested that DDD presumably exerted regulation through biological processes such as cholesterol metabolism and inflammatory response, and Kyoto Encyclopedia of Genes and Genomes(KEGG) analysis suggested that signaling pathways included lipid and atherosclerosis, insulin resistance, chemo-carcinogenesis-receptor activation, and AGE-RAGE signaling pathways in diabetic complications. In vitro experiments showed that DDD could reduce free fatty acid-induced lipid accumulation and cholesterol ester content in L02 cells and improve cellular activity, which might be related to the up-regulation of the expression of PPARα, LPL, PPARG, VEGFA, CETP, CYP1A1, and CYP3A4, and the down-regulation of the expression of TNF-α and IL-6. DDD may play a role in preventing and treating AS and HLP by improving lipid metabolism and inflammatory response, and inhibiting apoptosis with multi-component, multi-target, and multi-pathway characteristics.

Early intervention with Didang decoction delays macrovascular lesions in diabetic rats through regulating AMP-activated protein kinase signaling pathway / 中国天然药物

The study aimed to investigate the intervening role of Didang decoction (DDD) at different times in macrovascular endothelial defense function, focusing on its effects on the AMP-activated protein kinase (AMPK) signaling pathway. The effects of DDD on mitochondrial energy metabolism were also investigated in rat aortic endothelial cells (RAECs). Type 2 diabetes were induced in rats by streptozotocin (STZ) combined with high fat diet. Rats were randomly divided into non-intervention group, metformin group, simvastatin group, and early-, middle-, late-stage DDD groups. Normal rats were used as control. All the rats received 12 weeks of intervention or control treatment. Western blots were used to detect the expression of AMP-activated protein kinase α1 (AMPKα1) and peroxisome proliferator-activated receptor 1α (PGC-1α). Changes in the intracellular AMP and ATP levels were detected with ELISA. Real-time-PCR was used to detect the mRNA level of caspase-3, endothelial nitric oxide synthase (eNOS), and Bcl-2. Compared to the diabetic non-intervention group, a significant increase in the expression of AMPKα1 and PGC-1α were observed in the early-stage, middle-stage DDD groups and simvastatin group (P < 0.05). The levels of Bcl-2, eNOS, and ATP were significantly increased (P < 0.05), while the level of AMP and caspase-3 were decreased (P < 0.05) in the early-stage DDD group and simvastatin group. Early intervention with DDD enhances mitochondrial energy metabolism by regulating the AMPK signaling pathway and therefore may play a role in strengthening the defense function of large vascular endothelial cells and postpone the development of macrovascular diseases in diabetes.

Effects of Early Intervention of Didang Decoction on AMP-activated Protein Kinase Signaling Pathway in Diabetic Rats / 中国中医药信息杂志

Objective To observe the effects of the intervention of Didang Decoction at different times on changes of AMPK signaling pathway related factors in macrovascular endotheliocytes of diabetic rats; To discuss the mechanism of mitochondria energy metabolism regulating the AMPK signaling pathway for macrovascular endothelial defense function. Methods Injection of STZ into the caudal vein and administration of high fat diet wer used to generate diabetic rat model. All rats were randomly divided into the following 7 groups: control, model, metformin, simvastatin, early-, middle-, and late-stage Didang Decoction group. Western blot was used to detect the expressions of APMKα1 and PGC-1α in rat aortic endothelial cells. Changes in the intracellular AMP and ATP levels were detected by ELISA. Real time fluorescence quantitative PCR was used to detected mRNA expressions of Caspase-3, eNOS, and Bcl-2 in tissue of thoracic aorta. Results Compared with the model group, the expressions of AMPKα1 and PGC-1α in the early-stage and middle-stage Didang Decoction group and simvastatin group increased (P<0.05); the gene expressions of Bcl-2, and eNOS significantly increased in the early-stage Didang Decoction group and simvastatin group (P<0.05), while the expressions of Caspase-3 decreased significantly (P<0.05). The expression of ATP increased significantly and the expression of AMP decreased significantly in the early-stage Didang Decoction group and simvastatin group (P<0.05), and the best effects were shown in the early-stage Didang Decoction group. Conclusion Early intervention of Didang Decoction can enhance energy metabolism in the mitochondria of macrovascular endothelial cells by regulating the AMPK signaling pathway, and then plays a role in strengthening the defense function of macrovascular endothelial cells.

Early intervention of Didang Decoction on macroangiopathy in type 2 diabetic rats and its mechanisms / 中草药

Objective: To observe the effects of the early intervention with Didang Decoction (DDD) on macroangiopathy in type 2 diabetic rats. Methods: The type 2 diabetic rat model was established using high-fat diet and Streptozotocin (STZ), and the rats were divided into control, model, Pioglitazone (2.7 mg/kg), Simvastatin (1.8 mg/kg), early-, mid-, and late-term DDD-intervene groups (ig administered with 3.24 g/kg DDD once daily, before 4 weeks, at the same time, and after 4 weeks of STZ administration, respectively), the rats in each group were administered until 24 weeks of STZ administration. The immunohistochemical and pathological changes of intercellular cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in the aorta were observed, and the expression of nuclear factor-kappa B (NF-κB) and matrix metalloproteinases-9 (MMP-9) in aorta was detected using Western blotting. Results: Compared with the model group, the ICAM-1 expression decreased in early- and mid-term DDD-intervene groups, and Simvastatin group (P < 0.05), the VCAM-1 expression decreased in early-term DDD-intervene and Simvastatin groups (P < 0.05), the protein expression of NF-κB and MMP-9 was lower in early- and mid-term DDD-intervene groups (P < 0.05), and it is the most obvious in early-term DDD-intervene group. Conclusion: The contents of ICAM-1 and VCAM-1 as well as the protein expression of NF-κB and MMP-9 in type 2 diabetic rats decrease after early-term DDD-intervene, which could regulate NF-κB signaling pathway to delay the development of diabetic macroangiopathy.

Effect of modif ied Didang Decoction on AngII in insulin resistance rats / 中华中医药杂志

Objective:To explore the effect of modifi ed Didang Decoction on AngII in insulin resistance rats.Methods:The insulin resistance rats were randomly divided into the control group,the model control group,the high,medium and low dosage of modif ied Didang decoction groups and Wendiya group.Radioimmunity assay was used to observe the effect of modif ied Didang Decoction on the AngII in rats.Results:Compared with the model control group,the AngII signifi cantly decreased in the medium and high dose of modifi ed Didang Decoction groups(P

Effect of modified Didang Decoction on expression of TIMP-1 and PAI-1mRNA in glomerulosclerosis rats / 中华中医药杂志

Objective:To discuss the influence of Didang Decoction on glomerulosclerosis.Methods:The glomerulosclerosis model was established by uninephrectom and injection of adriamyci,and was treated by modif ied Didang Decoction and taken western medicine Losartan as the comparison group.To observe 24-hour urinary protein and kidney function of model group and each treatment group.Reverse transcriptase Polymerase Chain Reaction(RT-PCR) was used to determine the expression of TIMP-1 and PAI-1 in each group of rats kidney tissue.Results:Didang Decoction could decline elevated 24-hour urinary protein and improve kidney function,and decline the expression of TIMP-1 and PAI-1mRNA in glomerulosclerosis rats.Conclusion:Didang decoction could delay renal failure,it may be related to the down-regulated expression of TIMP-1 and PAI-1mRNA.