Exploration of the potential mechanism of Duhuo Jisheng Decoction in osteoarthritis treatment by using network pharmacology and molecular dynamics simulation.

In this study, the active ingredients of 15 Chinese herbal medicines of Duhuo Jisheng Decoction and their corresponding targets were obtained from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database. The microarray data of Osteoarthritis (OA) were obtained through the GEO database for differential analysis and then a drug target-OA-related gene protein-protein interaction (PPI) network was established. The potential targets of Duhuo Jisheng Decoction in the treatment of OA were acquired by intersecting the OA-associated genes with the target genes of active ingredients. Random walk with restart (RWR) analysis of PPI networks was performed using potential targets as seed, and the top 50 genes of affinity coefficients were used as key action genes of Duhuo Jisheng Decoction in the treatment of OA. A drug-active ingredient-gene interaction network was established. AKT1, a key target of Duhuo Jisheng Decoction in the treatment of OA, was obtained by topological analysis of the gene interaction network. Molecular docking and molecular dynamics verified the binding of AKT1 to its corresponding drug active ingredients. CETSA assay demonstrated that the combination of luteolin and AKT1 increased the stability of AKT1, and the combination efficiency was high. In conclusion, the molecular mechanism of Duhuo Jisheng Decoction in treating OA featured by multiple components, targets, and pathways had been further investigated in this study, which is of significance for discovering as well as developing new drugs for this disease. The findings can also offer personalized diagnosis and treatment strategies for patients with OA in clinical practice.

Duhuo Jisheng decoction alleviates neuroinflammation and neuropathic pain by suppressing microglial M1 polarization: a network pharmacology research.

BACKGROUND: Neuropathic pain (NP) is the most prevalent form of chronic pain resulting from nerve damage or injury. Despite the widespread use of Duhuo Jisheng decoction (DHJSD) in traditional Chinese medicine (TCM) to treat chronic pain, the mechanism underlying its analgesic action remains unclear. METHODS: Using network pharmacology, we obtained DHJSD and NP-related target information from public databases to construct protein-protein interactions (PPI) and compound-target networks based on common target genes. These networks were further analyzed using gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG). The interaction between molecules was verified through molecular docking using AutoDock Tools software. Additionally, we treated a chronic constriction injury (CCI) rat model with DHJSD and determined the mechanical withdrawal threshold (MWT). We used an enzyme-linked immunosorbent assay kit to determine the levels of inflammatory cytokines. Furthermore, qRT-PCR was employed to analyze ACHE, NOS2, MAPK3, PTGS2, AKT1, and PPARG mRNA expression, and immunofluorescence was used to evaluate changes in microglia. RESULTS: Our screening of compounds and targets identified 252 potential targets of DHJSD associated with NP. PPI analysis, along with GO and KEGG analyses, revealed that the potential mechanism of DHJSD in NP treatment may be related to inflammatory reactions, the IL-17 signaling pathway, MAP kinase activity, and endocrine activity. Based on molecular docking, the core target showed significant affinity for DHJSD's active components. Moreover, DHJSD treatment repaired the CCI-induced inflammatory reaction in the spinal cord while regulating the expression of ACHE, NOS2, MAPK3, PTGS2, AKT1, and PPARG mRNA. Immunofluorescence results indicated that the active components of DHJSD may regulate microglial M1 polarization to improve neuroinflammation, PPARG may have been involved in the process. CONCLUSION: The multi-component, multi-target, and multi-pathway actions of DHJSD provide new insights into its therapeutic mechanism in NP.

A review of Duhuo Jisheng decoction mechanisms in intervertebral disc degeneration in vitro and animal studies.

Intervertebral disc degeneration (IVDD) has become a serious public health problem, placing a heavy burden on society and the healthcare system. Its pathogenesis is not completely clear and may be closely related to mechanical damage, inflammatory factors, oxidative stress and death of nucleus pulposus cells (NPCs). The treatment of IVDD mainly includes conservative treatment and surgery. Conservative treatment is based on hormonal and anti-inflammatory drugs and massage techniques, which can relieve the pain symptoms to a certain extent, but cannot solve the problem from the root cause. Surgical treatment is mainly by removing the herniated nucleus pulposus, but it is more traumatic for IVDD patients, expensive and not suitable for all patients. Therefore, it is extremely important to clarify the pathogenesis of IVDD, to find an effective and convenient treatment and to further elaborate its mechanism of action. The effectiveness of traditional Chinese medicine in the treatment of IVDD has been well demonstrated in clinical medical research. We have been working on the Chinese herbal formula Duhuo Jisheng Decoction, which is a common formula for the treatment of degenerative disc disease. Not only does it have significant clinical effects, but it also has few adverse effects. At present, we found that its mechanism of action mainly involves regulation of inflammatory factors, reduction of apoptosis and pyroptosis of NPCs, inhibition of extracellular matrix degradation, improvement of intestinal flora, etc. However, a few relevant articles have yet comprehensively and systematically summarized the mechanisms by which they exert their effect. Therefore, this paper will comprehensively and systematically explain on it. This is of great clinical significance and social value for elucidating the pathogenesis of IVDD and improving the symptoms of patients, and will provide a theoretical basis and scientific basis for the treatment of IVDD with traditional Chinese medicine.

System pharmacology analysis to decipher the effect and mechanism of active ingredients combination from Duhuo Jisheng decoction on osteoarthritis in rats.

ETHNOPHARMACOLOGY RELEVANCE: Duhuo Jisheng decoction is a traditional Chinese formula that has been widely used in clinical practice to treat osteoarthritis, which has the effects of removing invaded cold and dampness, relieving joint pain. However, it is difficult to determine the effective substances and mechanisms due to assorted herbs and components, and further research is needed. AIM OF THE STUDY: This study was designed to explore and verify the mechanism and targets of DHJSD in the treatment of OA via network analysis and experiments. METHOD: In this study, the active ingredients of DHJSD were qualitatively analyzed by UPLC-QDA. Network analysis was used to identify common targets and pathways. Next, we explored the therapeutic mechanism of DHJSD through a rat model of knee osteoarthritis. HE staining was used to judge the establishment of the animal model. ELISA and Western blotting were used to verify the expression of key pathway proteins. CONCLUSION: In this study, seventeen chemical constituents in DHJSD were identified. According to the network analysis, we obtained the potential associated pathways of action. Then, molecular docking and SPR experiments showed that the sixteen identified components had high binding energies to IL-6. HE staining showed that the high-dose group of DHJSD had an obvious therapeutic effect on model rats. Compared with the model group, the levels of IL-1ß, TNF-α, IL-6, MMP3, MMP13, ADAMTS4 and ADAMTS5 in serum and the expression of STAT3 and p-STAT3 protein in administration groups were significantly decreased. This result indicated that the IL-6/STAT3 signaling pathway was one of the important pathways regulated by DHJSD to improve OA.

Duhuo Jisheng Decoction suppresses apoptosis and mitochondrial dysfunction in human nucleus pulposus cells by miR-494/SIRT3/mitophagy signal axis.

BACKGROUND: Increasing evidence suggests that mitophagy is responsible for the pathogenesis of intervertebral disk (IVD) degeneration. Previous studies have shown that Duhuo Jisheng Decoction (DHJSD), a classic Fangji of traditional Chinese medicine, can delay IVD degeneration; however, its specific mechanism of action is unknown. In this study, we investigated the mechanism by which DHJSD treatment prevented IVD degeneration in IL-1ß-treated human nucleus pulposus (NP) cells in vitro. METHODS: Cell Counting Kit-8 was performed to explore the effects of DHJSD on the viability of NP cells exposed to IL-1ß. The mechanism by which DHJSD delays IVD degeneration was explored using luciferase reporter assay, RT-qPCR, western blotting, TUNEL assay, mitophagy detection assay, Mito-SOX, Mitotracker and in situ hybridization. RESULTS: We observed that DHJSD enhanced the viability of NP cells treated with IL-1ß in a concentration-time dependent approach. Moreover, DHJSD lessened IL-1ß-induced NP apoptosis and mitochondrial dysfunction and activated mitophagy in NP cells treated with IL-1ß. Mitophagy suppressor cyclosporin A reversed the beneficial impacts of DHJSD in NP cells. In addition, the differential expression of miR-494 regulated IL-1ß-induced NP apoptosis and mitochondrial dysfunction, and the protective impact of miR-494 on NP cells treated with IL-1ß was achieved by mitophagy activation, which was regulated by its target gene, sirtuin 3 (SIRT3). Finally, we observed that DHJSD treatment could effectively delay IL-1ß-induced NP apoptosis by affecting the miR-494/SIRT3/mitophagy signal axis. CONCLUSIONS: These results show that the miR-494/SIRT3/mitophagy signaling pathway is responsible for the apoptosis and mitochondrial dysfunction of NP cells and that DHJSD may exert protective effects against IVD degeneration by regulating the miR-494/SIRT3/mitophagy signal axis.

Efficacy and safety of Duhuo Jisheng decoction combined with Western medicine in the treatment of ankylosing spondylitis: A systematic review and meta-analysis.

BACKGROUND: and purpose: The effects of Duhuo Jisheng Decoction (DJD) on ankylosing spondylitis (AS) remain controversial. This study aimed to assess the efficacy and safety of DJD combined with Western medicine in treating AS. METHODS: A total of nine databases were searched from the establishment of the databases to August 13th, 2021, for randomized controlled trials (RCTs) concerning the use of DJD combined with Western medicine to treat AS. Review Manager was used for the meta-analysis of the retrieved data. The risk of bias was evaluated using the revised Cochrane risk of bias tool for RCTs. RESULTS: The results indicated that the combinational use of DJD and Western medicine resulted in significantly higher outcomes in terms of effective rate (RR = 1.40, 95% CI: 1.30, 1.51); thoracic mobility (MD = 0.32, 95% CI: 0.21, 0.43); morning stiffness time (SMD = -0.38, 95% CI: 0.61, -0.14); BASDAI (MD = -0.84, 95% CI: 1.57, -0.10); VAS for pain [spinal (MD = -2.76, 95% CI: 3.10, -2.42); peripheral joint (MD = -0.84, 95% CI: 1.16, -0.53)]; CRP (MD = -3.75, 95% CI: 6.36, -1.14); ESR: (MD = -4.80, 95% CI: 7.63, -1.97); and adverse reactions (RR = 0.50, 95% CI: 0.38, 0.66) in comparison to the Western medicine alone in treating AS. CONCLUSION: Compared to the use of Western medicine, DJD combined with Western medicine improves the effective rate, functional scores, and symptoms of AS patients, with a reduced rate of adverse reactions.

Clinical effect of Huangqi Chifeng Decoction combined with modified Duhuo Jisheng Decoction in the treatment of ischemic stroke during recovery / 国际中医中药杂志

Objective:To evaluate the clinical curative effect of Huangqi Chifeng Decoction combined with modified Duhuo Jisheng Decoction on patients with ischemic stroke (IS) during recovery.Methods:Prospective cohort study. A total of 220 patients with IS during recovery who met inclusion criteria in the First People's Hospital of Dongcheng District in Beijing, were enrolled and divided into control group ( n=110) and observation group ( n=110) by random number table method between January 2015 and July 2020. The control group was given basic treatment, while observation group was given Huangqi Chifeng Decoction combined with modified Duhuo Jisheng Decoction. All were treated for 1 month. Before and after treatment, Traditional Chinese Medicine (TCM) syndromes were scored. The activities of daily living were evaluated by Barthel index. The quality of life was evaluated by stroke specific quality of life scale (SS-QOL). The severity of neurological impairment was evaluated by National Institute of Health Stroke Scale (NIHSS). The functional recovery of stroke was evaluated by modified Rankin scale (mRS). The adverse reactions during treatment were observed and recorded. Results:After treatment, scores of TCM syndromes, mRS and NIHSS in observation group were significantly lower than those in the control group ( t=21.87, 4.66, 12.06, P<0.01), while scores of Barthel index and SS-QOL were significantly higher than those in the control group ( t=14.13, 5.80, P<0.01). During treatment, there were no obvious adverse reactions in either group. Conclusion:Huangqi Chifeng Decoction combined with modified Duhuo Jisheng Decoction can improve clinical symptoms and nerve function in patients with IS during recovery, which is beneficial to improve outcomes and quality of life.

Network Pharmacological Study on Mechanism of the Therapeutic Effect of Modified Duhuo Jisheng Decoction in Osteoporosis.

Background: Modified Duhuo Jisheng Decoction (MDHJSD) is a traditional Chinese medicine prescription for the treatment of osteoporosis (OP), but its mechanism of action has not yet been clarified. This study aims to explore the mechanism of MDHJSD in OP through a combination of network pharmacology analysis and experimental verification. Methods: The active ingredients and corresponding targets of MDHJSD were acquired from the Traditional Chinese Medicine System Pharmacology (TCMSP) database. OP-related targets were acquired from databases, including Genecards, OMIM, Drugbank, CTD, and PGKB. The key compounds, core targets, major biological processes, and signaling pathways of MDHJSD that improve OP were identified by constructing and analysing the relevant networks. The binding affinities between key compounds and core targets were verified using AutoDock Vina software. A rat model of ovariectomized OP was used for the experimental verification. Results: A total of 100 chemical constituents, 277 targets, and 4734 OP-related targets of MDHJSD were obtained. Subsequently, five core components and eight core targets were identified in the analysis. Pathway enrichment analysis revealed that overlapping targets were significantly enriched in the tumour necrosis factor-alpha (TNF-α) signaling pathway, an inflammation signaling pathway, which contained six of the eight core targets, including TNF-α, interleukin 6 (IL-6), transcription factor AP-1, mitogen-activated protein kinase 3, RAC-alpha serine/threonine-protein kinase, and caspase-3 (CASP3). Molecular docking analysis revealed close binding of the six core targets of the TNF signaling pathway to the core components. The results of experimental study show that MDHJSD can protect bone loss, inhibit the inflammatory response, and downregulate the expression levels of TNF-α, IL-6, and CASP3 in ovariectomized rats. Conclusion: The mechanism of MDHJSD in the treatment of OP may be related to the regulation of the inflammatory response in the bone tissue.

Clinical study on thermal moxibustion combined with Duhuo Jisheng Decoction for the patients with knee osteoarthritis and liver and kidney deficiency syndrome / 国际中医中药杂志

Objective:To explore the clinical efficacy of thermal moxibustion combined with Duhuo Jisheng Decoction in the treatment of liver and kidney deficiency syndrome of knee osteoarthritis (KOA).Methods:From January 2020 to January 2021, 124 KOA patients with liver and kidney deficiency syndrome, who met the inclusion criteria, were divided into 2 groups according to the random number table method, with 62 in each group. The control group was treated with Duhuo Jisheng Decoction, and the observation group was treated with thermal moxibustion on the basis of the control group. Both groups were treated for 28 days. TCM symptom scores were performed before and after treatment, the Osteoarthritis Index of Western Ontario and McMaster University (WOMAC) was used to evaluate joint function. ELISA was used to detect serum insulin-like growth factor-1 (IGF-1), fibroblast growth factor-2 (FGF-2), transforming growth factor-β1 (TGF-β1), IL-1β, IL-6, TNF-α levels, and the clinical efficacy was evaluated.Results:The total effective rate was 91.9% (57/62) in the observation group and 77.4% (48/62) in the control group, and there was significant difference between two groups ( χ2=5.04, P=0.025). After treatment, the TCM symptom score and WOMAC score of the observation group were significantly lower than those of the control group ( t values were 11.33 and 12.23, respectively, all Ps<0.01). After treatment, the serum levels of IGF-1 [(15.63±2.03) ng/L vs. (12.78±1.57) ng/L, t=8.75], FGF-2 [(30.26±5.37) ng/L vs. (26.31±1.94) ng/L, t=5.45] and TGF-β1[(30.39±6.71)μg/L vs. (24.31±5.12) μg/L, t=5.67] in the observation group were significantly higher than those in the control group ( P<0.01), while the levels of IL-1β [(12.50±3.36) ng/L vs. (16.09±4.90) ng/L, t=4.76], IL-6 [(10.59±3.28) ng/L vs. (21.75 ± 4.09) ng/L, t=16.76] and TNF-α [(4.04±1.92) ng/L vs. (6.48±1.43) ng/L, t=8.03] in the observation group were significantly lower than those in the control group ( P<0.01). There was no adverse events from both groups during treatment. Conclusion:Thermal moxibustion combined with Duhuo Jisheng Ddecoction can alleviate the joint pain of KOA patients with liver and kidney deficiency syndrome, promote cartilage repair and improve the clinical curative effect.

Efficacy and safety of Duhuo Jisheng Decoction add-on bisphosphonate medications in patients with osteoporosis: A meta-analysis of randomized controlled trials.

ETHNOPHARMACOLOGICAL RELEVANCE: Duhuo Jisheng Decoction (DHJSD) is the most frequently prescribed herbal formula for the treatment of osteoporosis. However, efficacy and safety of DHJSD add-on bisphosphonate medications remain unclear. AIM OF THE STUDY: The purpose of this study was to reveal efficacy and safety of DHJSD add-on bisphosphonate medications in patients with osteoporosis through a systematic review with meta-analysis of randomized controlled trials (RCTs). METHODS: Five important databases were searched for RCTs on this topic, and two authors individually extracted information and data concerning study design, baseline characteristics, efficacy rate, bone mineral density (BMD), pain score, and adverse event. Meta-analysis was done mainly with risk ratio (RR) and standardized mean difference (SMD) for BMD and pain, using random-effects model; while Peto odds ratios (PORs) were used for pooling adverse event rates due to sparse data. Point estimate was reported with 95% confidence intervals (CIs). RESULTS: Seventeen RCTs (n = 1526) met eligibility criteria, and were included in this synthesis. Pooled estimates demonstrated that as compared with no DHJSD, DHJSD-B led to significantly higher efficacy rates (RR = 1.25, 95%CI: 1.19-1.31; I2 = 0%), more lumbar BMD (SMD = 0.61, 95%CI: 0.25-0.96; I2 = 20%), lower pain score (SMD = -1.10, 95%CI: 1.40-0.79; I2 = 33%), and lower overall adverse event rates (POR = 0.40; 95%CI: 0.20-0.97; I2 = 27%). CONCLUSION: Adding DHJSD on bisphosphonate medications seems to be an effective and safe strategy in treating patients with osteoporosis.

Clinical study on modified Duhuo Jisheng Decoction combined with Traditional Chinese Medicine fumigation in the treatment of knee osteoarthritis / 国际中医中药杂志

Objective:To explore the effects of modified Duhuo Jisheng Decoction combined with Traditional Chinese Medicine (TCM) fumigation on joint function and inflammatory mediators in synovial fluid of patients with knee osteoarthritis.Methods:A total of 203 patients with knee osteoarthritis who met the inclusion criteria between March 2020 and March 2021 in the hospital were divided into observation group (102 cases) and control group (101 cases) according to the random number table method. The control group was given oral administration of celecoxib capsules + TCM fumigation, and the observation group was given modified Duhuo Jisheng Decoction on the basis of the control group. Both groups were treated for 3 months. TCM symptoms were scored before and after treatment, and Japanese Orthopaedic Association scores (JOA) was used to evaluate the knee function. Immunoturbidimetry was adopted to detect serum high-sensitivity C-reactive protein (hs-CRP), and ELISA was used for the detection of levels of TNF-α and IL-6 in synovial fluid, and the clinical efficacy was evaluated.Results:The total effective rate was 90.2% (92/102) in observation group and was 79.2% (80/101) in control group ( χ2=4.74, P=0.030). The JOA scores at 1 month and 3 months after treatment in observation group were significantly higher than those in the control group ( t=3.17, 7.74, all Ps<0.01). The scores of cold and painful knee joint, soreness and weakness of waist and knees, joint swelling and inhibited bending and stretching were significantly in the observation group after treatment lower than those in the control group ( t=7.61, 10.81, 6.62, 8.77, all Ps<0.001). The levels of TNF-α and IL-6 in synovial fluid and level of serum hs-CRP were significantly in the observation group after treatment lower than those in the control group ( t=7.97, 9.52, 9.56, all Ps<0.001). During treatment, the incidence rate of adverse reactions was 32.7% (33/101) in control group and that of observation group was 19.6% (20/102) ( χ2=4.49, P=0.034). Conclusion:Modified Duhuo Jisheng Decoction combined with TCM fumigation can improve the clinical symptoms and knee function, relieve the inflammatory response and enhance the clinical efficacy of patients with knee osteoarthritis.

Duhuo jisheng decoction suppresses matrix degradation and apoptosis in human nucleus pulposus cells and ameliorates disc degeneration in a rat model.

ETHNOPHARMACOLOGICAL RELEVANCE: The lower back pain (LBP) caused by intervertebral disc (IVD) degeneration brings a heavy burden to society. A classic treatment method of Chinese medicine, fangji-duhuo jisheng decoction (DHJSD), has been effective in the clinical treatment of LBP, although the underlying mechanism remains unknown. AIM OF THE STUDY: In this work, the main objective was to study the effects of DHJSD on in vitro IVD degeneration of human nucleus pulposus (NP) cells after pressure treatment and on an in vivo interrupted IVD degeneration rat model. MATERIALS AND METHODS: The effects of DHJSD on the viability of NP cells were detected using Cell Counting Kit-8. RT-qPCR, western blotting, TUNEL assay, transmission electron microscopy, and immunofluorescence staining were performed to explore the molecular mechanism underlying protection against compression-induced matrix degradation and apoptosis in NP cells by DHJSD. Furthermore, the effects of DHJSD on IVD degeneration in a rat IDD model were also determined. RESULTS: We found that DHJSD increased the viability of NP cells in a concentration- and time-dependent manner. Furthermore, DHJSD significantly reduced compression-induced NP matrix degeneration and apoptosis, activated autophagy, and inhibited the p38/MAPK signaling pathway in NP cells subjected to compression. Autophagy inhibitor 3-MA and p38/MAPK signaling pathway activator anisomycin reversed the beneficial effects of DHJSD in NP cells, indicating that DHJSD protects against IVD degeneration by autophagy activation and P38/MAPK signaling pathway inhibition. Furthermore, DHJSD treatment effectively delayed IVD degeneration in a puncture-induced IDD rat model. CONCLUSIONS: DHJSD prevents compression-induced matrix degradation and cell apoptosis through regulating autophagy and the P38/MAPK signaling pathway. The mechanism underlying the effects of DHSJD elucidated in this study provides a new direction for LBP treatment.