1.Comparative Study on Determination of Astragaloside, AstragalosideⅠ, AstragalosideⅡ and AstragalosideⅢ in Mongolia Radix Astragali from Various Habitats
Jingchao SHI ; Haozheng LI ; Yonghui WANG ; Ran ZHOU ; Biwang LIU
World Science and Technology-Modernization of Traditional Chinese Medicine 2015;(5):1016-1021
The HPLC-ELSD method was used in the content determination of astragaloside, astragalosideⅠ, astragalosideⅡand astragalosideⅢ in Mongolia Radix Astragali (Astragalus membranaceus(Fisch.) Bge. var. mongholicus(Bge.) Hsiao) among 16 batches from various habitats. The DIKMA Diamonsil C18 (150 mm× 4.6 mm, 5μm) was adopted with acetonitrile and water as the mobile phase at a gradient mode program. The flow rate was 1.0 mL·min-1. And the column temperature was 30℃. The ELSD detector parameters were the drift tube temperature at 90℃, and the air flow rate of 2.8 L·min-1. The SPSS 16.0 software was used in the cluster analysis of content determination. The results showed that when the injection volume was within the range of 0.093 2-1.02μg (r = 0.999 5), 0.789-8.78μg (r = 0.999 7), 0.506-3.13μg (r = 0.999 6), and 0.016 1-1.38μg (r = 0.999 2) for astragaloside, astragalosideⅠ, astragalosideⅡ and astragalosideⅢ, respectively, the average recoveries were 97.55%, 98.61%, 99.68%, 98.58%with RSD of 1.2%, 1.3%, 1.3%, 1.2%, respectively. The results of cluster analysis showed that the single using of astragaloside as index was unable to differentiate Mongolia Radix Astragali from various habitats. However, the simultaneous determination of 4 types of astragalosides as indexes can differentiate Mongolia Radix Astragali from various habitats. It was concluded that the method was simple, quick and accurate, which can directly reflect the quality status of Mongolia Radix Astragali from different origins. It also provided new ideas for the quality control of Mongolia Radix Astragali.
2.Improving mechanism of Buyang huanwu decoction on idiopathic pulmonary fibrosis model rats
China Pharmacy 2023;34(14):1671-1675
OBJECTIVE To study the improving mechanism of Buyang huanwu decoction (BYHW) on idiopathic pulmonary fibrosis (IPF) model rats. METHODS Ninety-six Wistar rats were randomly divided into blank group, model group, positive control group, and BYHW low-dose, middle-dose and high-dose groups, with 16 rats in each group. Except for the blank group, the IPF model was induced in other groups by intratracheal instillation of bleomycin (5 mg/kg). Starting from the day of modeling, the blank group and model group were given normal saline (10 mL/kg) intragastrically, while the rats in the positive control group were given dexamethasone solution (5 mg/kg) intragastrically, BYHW low-dose, middle-dose and high-dose groups were treated with BYHW (2.5, 5, 10 g/kg, by crude drug) intragastrically, once a day, for 28 consecutive days. The body mass of rats in each group was measured on days 0, 7, 14, 21 and 28 after modeling. The number of adherent white blood cells in pulmonary veins was observed by a dynamic visualization system. The contents of TNF-α and IL-6 in serum and TNF-α, IL-6, HYP and TGF-β1 in lung tissue were detected; the protein expression of ZO-1 was also detected. The pathomorphological changes in lung tissue were observed. RESULTS Compared with the model group, the body weight of rats all increased significantly in BYHW high-dose and middle-dose groups, positive control group, while the number of adherent white blood cells in pulmonary veins was decreased significantly; the contents of TNF-α (except for serum in BYHW middle-dose group) and IL-6 in serum and lung tissue, the contents of HYP and TGF-β1 in lung tissue were decreased significantly, while the protein expression of ZO-1 in the lung tissue was increased significantly (P<0.05 or P<0.01). The pathological changes of lung tissue were improved to varying degrees. CONCLUSIONS BYHW may play anti-pulmonary fibrosis role by improving leukocyte adhesion, anti-inflammatory, anti-fibrosis, and other aspects of pulmonary microcirculation.
3.Danggui Sinitang Mitigates Gouty Arthritis in Rats by Regulating Autophagy via PI3K/Akt/mTOR Signaling Pathway
Rongrong LU ; Yue CAO ; Xiaolei LEI ; Ruchao LI ; Biwang LIU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(12):78-84
ObjectiveTo study the mechanism of Danggui Sinitang in mitigating gouty arthritis (GA) in rats by regulating autophagy via the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway. MethodSixty male SD rats were randomly assigned into normal, model, colchicine (0.3 mg·kg-1), and low-, medium-, and high-dose Danggui Sinitang (6.54, 13.08, and 26.16 g·kg-1) groups (n=10) and administrated with corresponding drugs by gavage. The rats in the normal group and model group were administrated with equal volume of normal saline by gavage for 7 days. One hour after administration on day 5, the GA model was established by injecting sodium urate suspension (50 g·L-1) into the right ankle joint of rats in other groups except the normal group, and the rats in the normal group were injected with sterile normal saline of the same volume. The swelling and pathological changes of the ankle joint were observed. The serum levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IL-1β were determined. Western blot was employed to determine the protein levels of PI3K, phosphorylated PI3K (p-PI3K), protein kinase B (Akt), phosphorylated Akt (p-Akt), mTOR, phosphorylated mTOR (p-mTOR), microtubule-associated protein 1 light chain 3 Ⅱ/Ⅰ (LC3Ⅱ/Ⅰ), autophagy effector Beclin-1, and ubiquitin-binding protein p62 in the synovial tissue. Real-time fluorescent quantitative PCR (Real-time PCR) was employed to determine the mRNA levels of PI3K, Akt, mTOR, LC3, Beclin-1 and p62. ResultCompared with the normal control, the model group showed increased joint swelling index (P<0.01), elevated serum levels of TNF-α, IL-6, and IL-1β, inflammatory cell infiltration, and fibrous tissue hyperplasia. In addition, the model group showed up-regulated protein levels of PI3K, p-PI3K, Akt, p-Akt, mTOR, p-mTOR, and p62 and mRNA levels of PI3K, Akt, mTOR, and p62 in the synovial tissue, while it showed down-regulated protein levels of LC3Ⅱ/Ⅰ and Beclin-1 and mRNA levels of LC3 and Beclin-1 (P<0.01). Compared with the model group, medium- and high-dose Danggui Sinitang alleviated the joint swelling (P<0.01), lowered the serum levels of TNF-α, IL-6, and IL-1β (P<0.05), and relieved the inflammatory cell infiltration in the synovial tissue of the ankle joint and the fibrous tissue hyperplasia. Moreover, they down-regulated the protein levels of PI3K, p-PI3K, Akt, p-Akt, mTOR, p-mTOR, and p62 and the mRNA levels of PI3K, Akt, mTOR, and p62 in the synovial tissue (P<0.05), while they up-regulated the protein levels of LC3Ⅱ/Ⅰ and Beclin-1 and the mRNA levels of LC3 and Beclin-1 (P<0.05). ConclusionDanggui Sinitang, especially at a high dose, can inhibit PI3K/Akt/mTOR signaling pathway to improve autophagy in the synovial tissue, thereby mitigating GA.
4.Treatment of Hyperuricemia and Gouty Arthritis by Buyang Huanwu Tongfeng Decoction via Inhibition of PPAR-γ/NF-κB/AGEs/RAGE Pathway Based on Network Pharmacology
Yue CAO ; Wanmei YAO ; Tao YANG ; Man YANG ; Ruimin JIA ; Rongrong LU ; Xue FENG ; Biwang LIU
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(1):182-192
ObjectiveThis paper aims to investigate the potential molecular biological mechanism of Buyang Huanwu Tongfeng decoction in treating hyperuricemia and gouty arthritis by network pharmacology and molecular docking technology and preliminarily verify the mechanism through animal experiments. MethodsThe active ingredients and targets in the Buyang Huanwu Tongfeng decoction were obtained by the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) and ETCM databases. The DisGeNET and GeneCards databases were utilized to acquire disease targets associated with hyperuricemia and gouty arthritis. These disease targets were then intersected with drug targets to identify key targets. The R language ClusterProfiler package and Python were employed for conducting gene ontology(GO) enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG) enrichment analysis. The regulatory network diagram of the drug-key target-function-pathway was visualized using Cytoscape 3.9.1 software, and the protein-protein interaction (PPI) network for key targets was depicted. Finally, the hub gene was determined through topological analysis. Auto Dock, PyMOL, and other software were used for molecular docking to explore the possible therapeutic mechanism of Buyang Huanwu Tongfeng decoction for hyperuricemia and gouty arthritis. In animal experiments, a composite rat model of hyperuricemia induced by intraperitoneal injection of oteracil potassium combined with gouty arthritis induced by the modified Coderre method was established. Through hematoxylin-eosin(HE) staining, uric acid test, enzyme linked immunosorbent assay(ELISA), Western blot, and real-time polymerase chain reaction(Real-time PCR), the molecular mechanism and key targets of Buyang Huanwu Tongfeng decoction for treating hyperuricemia and gouty arthritis were observed. ResultsAfter screening and removing duplicate values, 76 active ingredients and 15 key targets were finally obtained. GO enrichment analysis yielded that the treatment of hyperuricemia and gouty arthritis with Buyang Huanwu Tongfeng decoction was significantly associated with acute inflammatory response, astrocyte activation, regulation of interleukin (IL)-8 production, nuclear receptor activity, and binding of growth factor receptor. KEGG pathway enrichment analysis obtained that the key target genes were significantly associated with the IL-17 signaling pathway, advanced glycosylation end/receptor of advanced glycation endproducts(AGE/RAGE) signaling pathway, anti-inflammatory, and other pathways. PPI network indicated that albumin(ALB), peroxisome proliferator-activated receptor-γ (PPAR-γ), IL-6, IL-1β, and C-reactive protein(CRP) were the key protein targets. The molecular docking results showed that ALB had the strongest binding force with beta-carotene (β-carotene). Biochemical results showed that blood uric acid decreased in the Buyang Huanwu Tongfeng decoction groups. HE staining results showed that the low-dose (7.76 g·kg-1·d-1), medium-dose (15.53 g·kg-1·d-1), and high-dose (31.05 g·kg-1·d-1) groups of Buyang Huanwu Tongfeng decoction had different degrees of remission, and the remission of the high-dose group was the most obvious. Fibroblastic tissue hyperplasia in synovial joints accompanied with inflammatory cell infiltration, as well as inflammatory cell infiltration in renal tissue of the high-dose group was significantly reduced, followed by the medium-dose and low-dose groups, and the expression of ALB, PPAR-γ, IL-6, IL-1β, and CRP was down-regulated to different degrees. ConclusionBy regulating the targets such as ALB, PPAR-γ, IL-6, IL-1β, and CRP, inhibiting the PPAR-γ/nuclear transcription factor (NF)-κB pathway, and reducing AGEs/RAGE-mediated inflammation, Buyang Huanwu Tongfeng decoction exerts anti-inflammatory and analgesic effects and activates blood circulation and diuresis in the treatment of hyperuricemia and gouty arthritis.