Treatment of Bone and Joint Diseases with Gancao Fuzitang： A Review / 中国实验方剂学杂志

Gancao Fuzitang originates from the Treatise on Febrile Diseases and Miscellaneous Diseases （《伤寒杂病论》） and is mainly used to treat pain in the bones and joints and symptoms such as no flexion or extension. It has the effect of tonifying the spleen and kidney and removing dampness and turbidity， so it is widely used in the clinical treatment of various bone and joint diseases. This article reviewed the clinical research and mechanism of Gancao Fuzitang in the treatment of bone and joint diseases. The research has found that this prescription has good efficacy in treating bone and joint diseases such as rheumatoid arthritis， rheumatoid arthritis， ankylosing spondylitis， gout， and intervertebral disc herniation. Its mechanism of action may be related to regulating the level of inflammatory factors， antioxidation， and the protein expression of inflammatory and apoptotic cell-related pathways， improving bone and joint diseases， and alleviating related symptoms. This study can provide a reference for further deepening the research on the prevention and treatment of bone and joint diseases with Gancao Fuzitang.

Virtual screening combined with experimental verification reveals the potential mechanism of Fuzitang decoction against Gouty Arthritis.

Background: and Purpose: Fuzitang decoction (FZT), a classic prescription of traditional Chinese medicine (TCM), has excellent efficacy in treating gouty arthritis (GA). However, the underlying molecular mechanism remains obscure. In the present study, we aimed to explore the underlying mechanisms of FZT in treating GA by virtual screening combined with experimental verification. Methods: In this study, the active components of FZT and their corresponding targets were screened from the TCMSP database and TargetNet database. Then, the potential targets of FZT against GA were retrieved from multiple databases to generate a network. Protein-protein interaction, herbal-component-target, Gene Ontology (GO) enrichment, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were applied to identify potential targets and related signaling pathways. Furthermore, molecular docking simulation was applied to identify the interactions between the drug and targets. Finally, in vitro experiments were conducted to validate the potential targets and signaling pathways. Results: In the present study, several crucial components, including kaempferol, luteolin, catechin, deoxyandrographolide, and perlolyrine in FZT, were obtained through network pharmacology, and several potential targets to treat GA were developed, such as PPARG, CYP3A4, PTGS2 (known as COX2), VEGFA, and CYP1A1. Experimental validation suggested that deoxyandrographolide significantly suppressed the expression of IL-1ß, COX2, NLRP3 and IL-6 in inflammatory monocyte cells. Conclusions: Our results identified a novel anti-inflammatory compound, deoxyandrographolide, which helps to explain the potential mechanism of FZT in treating GA and provides evidence to support FZT's clinical use.

Effect of Fuzitang on Proliferation of Human Rheumatoid Arthritis Synovial Fibroblast Cell Line MH7A and Expression of miR-155 / 中国实验方剂学杂志

ObjectiveTo observe the effects of Fuzitang （FZT） on the proliferation of MH7A cells， the human rheumatoid arthritis synovial fibroblasts， and the expression of miR-155 and explore its anti-rheumatoid arthritis mechanism. MethodMH7A cells were cultured in vitro and divided into a blank group， high- （25 g·L-1） and low-dose （12.5 g·L-1） FZT groups， and a positive drug group （hydroxychloroquine， 0.006 25 g·L-1）. The cell proliferation was detected by cell counting kit-8（CCK-8） method， and the change in the MH7A cell cycle was detected by flow cytometry. The mRNA expression of miR-155 and its downstream genes， including SH2 domain-containing inositol 5-phosphatase-1（SHIP-1）， protein kinase B 3（Akt3）， and mammalian target of rapamycin（mTOR）， was detected by Real-time fluorescence quantitative polymerase chain reaction（Real-time PCR）， and the protein expression of phosphatidylinositol 3-kinase （PI3K）， Akt3， and mTOR was detected by Western blot. ResultFZT in vitro in a concentration of 6.25 g·L-1 above could inhibit the proliferation of MH7A cells in the significant dose- and time-effect manner. Compared with the blank group， the FZT groups showed increased proportions of cells in the G2/M phase （P<0.05）， and the high-dose FZT group showed a decreased proportion of cells in the G0/G1 phase （P<0.05）. The arresting effect of FZT on the cell cycle was in a significant dose-effect manner. Compared with the blank group， the FZT groups showed down-regulated miR-155 and mTOR mRNA expression （P<0.05）， and the high-dose FZT group showed up-regulated SHIP1 mRNA expression and down-regulated Akt3 mRNA expression （P<0.05）. Compared with the blank group， the FZT groups showed reduced protein expression of PI3K， Akt3， and mTOR （P<0.05）. ConclusionFZT can significantly inhibit the proliferation of MH7A cells， and the mechanism is related to the promotion of the expression of SHIP-1 and down-regulation of the gene expression of the PI3K/Akt3/mTOR signaling pathway by down-regulating the expression of miR-155.

Clinical Effect of Mahuang Xixin Fuzitang Combined with Acupuncture and Moxibustion in Treatment of Localized Scleroderma / 中国实验方剂学杂志

ObjectiveTo observe the clinical effect of Mahuang Xixin Fuzitang combined with acupuncture and moxibustion in the treatment of localized scleroderma. MethodA total of 95 patients with localized scleroderma treated in Wuhan No. 1 Hospital from September 2019 to October 2021 were assigned into a control group （47 patients） and an observation group （48 patients） by random number table method. The control group was treated with Centella triterpenes tablets and heparin sodium cream， and the observation group was additionally treated with Mahuang Xixin Fuzitang combined with acupuncture and moxibustion. Both groups were treated for 8 weeks， and the clinical effect was compared between groups. The traditional Chinese medicine（TCM） syndrome score （local skin sclerosis， loss of skin texture， darkening of skin pigment， scaly dry skin， etc.）， serum levels of soluble interleukin-2 receptor （sIL-2R） and tumor necrosis factor-alpha （TNF-α）， erythrocyte sedimentation rate （ESR）， and eosinophil count （EO） were compared between before and after treatment as well as between groups. Additionally， the adverse reactions were recorded. ResultThe observation group had higher total effective rate than the control group ［95.83% （46/48） vs. 82.98% （39/47）， χ2=4.166 4， P<0.05］. Before treatment， the TCM syndrome score， sIL-2R， TNF-α， ESR， and EO showed no significant differences between the two groups. The 8 weeks of treatment improved the TCM syndrome score， sIL-2R， TNF-α， ESR， and EO. Moreover， the observation group was superior to the control group in these indicators （P<0.05）. During the treatment， the observation group showed 1 case of abnormal liver function and 1 case of nausea and vomiting， and the control group had 1 case of nausea and vomiting， 1 case of abnormal renal function， and 1 case of abnormal liver function. The total adverse reactions of the observation group （4.17%） and the control group （6.38%） had no significant difference （χ2=0.233 9， P=0.062 86）. ConclusionMahuang Xixin Fuzitang combined with acupuncture and moxibustion is safe and effective in the treatment of localized scleroderma.

Clinical Effect of Mahuang Xixin Fuzitang Combined with Acupuncture and Moxibustion in Treatment of Localized Scleroderma / 中国实验方剂学杂志

ObjectiveTo observe the clinical effect of Mahuang Xixin Fuzitang combined with acupuncture and moxibustion in the treatment of localized scleroderma. MethodA total of 95 patients with localized scleroderma treated in Wuhan No. 1 Hospital from September 2019 to October 2021 were assigned into a control group （47 patients） and an observation group （48 patients） by random number table method. The control group was treated with Centella triterpenes tablets and heparin sodium cream， and the observation group was additionally treated with Mahuang Xixin Fuzitang combined with acupuncture and moxibustion. Both groups were treated for 8 weeks， and the clinical effect was compared between groups. The traditional Chinese medicine（TCM） syndrome score （local skin sclerosis， loss of skin texture， darkening of skin pigment， scaly dry skin， etc.）， serum levels of soluble interleukin-2 receptor （sIL-2R） and tumor necrosis factor-alpha （TNF-α）， erythrocyte sedimentation rate （ESR）， and eosinophil count （EO） were compared between before and after treatment as well as between groups. Additionally， the adverse reactions were recorded. ResultThe observation group had higher total effective rate than the control group ［95.83% （46/48） vs. 82.98% （39/47）， χ2=4.166 4， P<0.05］. Before treatment， the TCM syndrome score， sIL-2R， TNF-α， ESR， and EO showed no significant differences between the two groups. The 8 weeks of treatment improved the TCM syndrome score， sIL-2R， TNF-α， ESR， and EO. Moreover， the observation group was superior to the control group in these indicators （P<0.05）. During the treatment， the observation group showed 1 case of abnormal liver function and 1 case of nausea and vomiting， and the control group had 1 case of nausea and vomiting， 1 case of abnormal renal function， and 1 case of abnormal liver function. The total adverse reactions of the observation group （4.17%） and the control group （6.38%） had no significant difference （χ2=0.233 9， P=0.062 86）. ConclusionMahuang Xixin Fuzitang combined with acupuncture and moxibustion is safe and effective in the treatment of localized scleroderma.

Clinical Research on Treatment of Acute Cerebral Infarction with ‘Sanjiao Regular Sequence Therapy’ / 中国实验方剂学杂志

Objective:To evaluate the clinical efficacy of Sanjiao Cidi therapy on acute cerebral infarction and its effect on levels of S100-β protein （S100-β）， tumor necrosis factor-α （TNF-α）， hypersensitive C-reactive protein （hs-CRP） and neuropeptide （NPY）. Method:One hundred and eighty patients were randomly divided into control group （90 cases） and observation group （90 cases） by random number table. Patients in control group got aspirin enteric-coated tablets， 100 mg/time， 1 time/day， edaravone injection （injected within 30 minutes） for 14 days， 30 mg/time， 2 times/day， simvastatin tablets， 20 mg/time， 1 time/day. In addition to the basic therapy of meloxicam tablets， patients in observation group were also treated with Sanjiao Cidi therapy. In the first step， patients got Guizhi therapy to dredge Zhongjiao and Shangjiao， 1 dose/day， for 8 days. In the second step， patients got Sini therapy to dredge Zhongjiao and Xiajiao， 1 dose/day， for 10 days. In the third step， patients got Tianjing Gubentherapy， 1 dose/day， for 10 days. The course of treatment was 4 weeks. Before the treatment， and at the first， second， third and fourth weeks after treatment， National Institutes of Health Stroke Scale （NIHSS） was scored. And before and after treatment， function scale of fuglmeyer （FMA）， ability of daily life activities （ADL）， mini-mental state examination （MMSE） and main symptoms of traditional Chinese medicine were scored. Comprehensive assessment of patient report outcome （PRO） was made. And levels of S100-β， hs-CRP， TNF-α and NPY were detected. And the incidence rate of pulmonary infection， urinary infection， skeletal myalgia， shoulder hand syndrome and shoulder subluxation of patients were recorded during hospitalization. Result:The clinical efficacy in observation group was better than that in control group （Z=2.141， P<0.05）. Scores of NIHSS in observation group were lower than those in control group at the first， second， third and fourth weeks after treatment （P<0.01）. Scores of upper limb， legs and the total scores from FMA were higher than those in control group （P<0.01）. Scores of the main symptoms of traditional Chinese medicine， symptoms， psychological and social scores， total scores of PRO， S100-β， hs-CRP， TNF-α and NPY were lower than those in the observation group （P<0.01）. And scores of ADL and MMSE were higher than those in control group （P<0.01）. Total incidence of complications in observation group was 27.27%（21/77）， which was lower than 46.15%（36/78） in control group （χ2=5.941， P<0.05）. Conclusion:In addition to conventional western medicine treatment， Sanjiao Cidi therapy can treat the patients with acute cerebral infarction， alleviate the degree of neurological deficit， improve the cognitive function， motor function of limbs and the ability of daily life， reduce the main symptoms of traditional Chinese medicine， the incidence of complications and the inflammatory response， protect the nerve cells， with a better clinical efficacy and comprehensive effect in patients than pure Western medicine.

Effect of Gancao Fuzitang on Synovial Fibroblast-like Cell Proliferation in Adjuvant Arthritis Mice Model / 中国实验方剂学杂志

Objective:To investigate the effect of Gancao Fuzitang on the proliferation of fibroblast-like synoviocytes (FLS) in synovial tissue of mice with adjuvant-induced arthritis (AA). Method:The 48 male Balb/c mice were randomly divided into 4 groups:normal control group, model group(arthritis induced through injection with adjuvant), Gancao Fuzitang group and triptolide group. The normal control group and the model control group were orally given physiological saline. After the successful modeling, the mice of the other two groups were treated with Gancao Fuzitang or triptolide orally for 18 days. Then the animals were euthanized, the paw edema of the animals was measured, and arthritic ankles were observed by histological examination. Tumor necrosis factor (TNF)-α was examined by immunoassays. The proliferation of synovial cells of foot joints was detected by immunohistochemical expressions of Vimentin. The expressions of Cyclin D1, proliferating cell nuclear antigen (PCNA), p53 and p21 were detected by Western blot. Result:Compared with the normal control group, the mice of synovial hyperplasia and bone erosion in model group were obvious. The joint swelling degree, TNF-α level and proliferation of FLS were increased (P1,PCNA were up-regulated(PPα, cell cycles (Cyclin D1 and PCNA), and increased protein expressions of p53 and p21 (PConclusion:Gancao Fuzitang has a therapeutic effect on AA mice, and the mechanism might be associated with its anti-inflammatory effect and the effects in regulating Cyclin D1, PCNA, p53 and p21 expressions and inhibiting FLS proliferation.

Effect of Wuzang Wenyang Huayu Decoction on Neurofibrillary Tangles by Inhibiting Cyclin-dependent Kinase-5 Over-expression in SAMP8 Mice / 中国实验方剂学杂志

Objective: To explore the mechanism of Wuzang Wenyang Huayu decoction in improving the cognitive competence and the pharmacological mechanism for neurofibrillary tangles related to cyclin-dependent kinase-5(CDK-5).Method: The 10 SAMR1 mice were used as normal group,40 SAMP8 mice were randomly divided into model group,donepezil group (0.4 mg·kg-1·d-1),high and low dose Wuzang Wenyang Huayu decoction groups (5,1.25 g·kg-1·d-1).Drugs were administered by gastric lavage for 4 continuous weeks.Directional navigation and space exploration ability were evaluated with Morris amaze.Real-time PCR was used to measure the mRNA expression of CDK-5 in brain nerve tissues.Western blot was used to detect the protein expression of CDK-5 and phosphorylation of Tau protein.Meanwhile,neurofibrillary tangles in brain tissue were detected with silver staining method.Result: As compared with normal group,both CDK-5 expression and Tau protein phosphorylation in brain nerve tissues were remarkably increased in model group (PPPPPPConclusion: Wuzang Wenyang Huayu decoction can markedly improve the cognitive competence of SAMP8 mice,and the mechanism may be related to its inhibition on CDK-5 over-expression,and down-regulation of Tau protein phosphorylation and neurofibrillary tangles in brain tissue.