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1.
PLoS One ; 19(6): e0305141, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38857250

RESUMO

To analyze the lower limb muscle strength characteristics of amateur runners with patellofemoral pain (PFP). Sixty amateur runners (30 in the knee pain group, 30 in the control group) underwent measurements of hip abduction strength, knee extension strength, and knee flexion strength. Additionally, the hamstring/quadriceps ratio and limb strength symmetry index were calculated for all participants. Statistical analyses were conducted using linear mixed models. The hip abduction and knee extensors strength of amateur runners with PFP was significantly lower than that of the control group. The hamstring/quadriceps ratio was significantly higher in the male knee pain group compared to the control group, while no significant difference was found between the female knee pain group and the control group. Furthermore, both the hip abduction strength symmetry index and knee extensors symmetry index were significantly lower in the knee pain group compared to the control group. Amateur runners with PFP exhibit distinct lower limb strength characteristics compared to non-knee pain runners. Additionally, differences in lower limb strength characteristics between male and female amateur runners with PFP were observed. These findings emphasize the importance of considering functional and gender differences in PFP rehabilitation training.


Assuntos
Extremidade Inferior , Força Muscular , Síndrome da Dor Patelofemoral , Corrida , Humanos , Masculino , Feminino , Força Muscular/fisiologia , Estudos Transversais , Corrida/fisiologia , Adulto , Síndrome da Dor Patelofemoral/fisiopatologia , Extremidade Inferior/fisiopatologia , Adulto Jovem
2.
Biomed Pharmacother ; 171: 116190, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38278026

RESUMO

Sinomenine (SIN), an alkaloid extracted from the Chinese herbal medicine Sinomenium acutum, has great potential in anti-inflammatory, immune regulation, analgesic and sedative, and is already a clinical drug for the treatment of rheumatoid arthritis in China. Our previous studies show SIN inhibits inflammation by regulating ɑ7nAChR, a key receptor of cholinergic anti-inflammatory pathway (CAP), which plays an important role in regulating peripheral and central nervous system inflammation. Growing evidence supports the cholinergic dysregulation and inflammatory responses play the key role in the pathogenesis of AD. The intervention effects of SIN on AD by regulating CAP and homeostasis in brain and gut were analyzed for the first time in the present study using scopolamine-induced AD model mice. Behavioral tests were used to assess the cognitive performance. The neurons loss, cholinergic function, inflammation responses, biological barrier function in the mouse brain and intestinal tissues were evaluated through a variety of techniques, and the gut microbiota was detected using 16SrRNA sequencing. The results showed that SIN significantly inhibited the cognitive decline, dysregulation of cholinergic system, peripheral and central inflammation, biological barrier damage as well as intestinal flora disturbance caused by SCOP in mice. More importantly, SIN effectively regulated CAP to suppress the activation of TLR4/NF-κB and protect the homeostasis in brain and gut to alleviate cognitive impairment.


Assuntos
Doença de Alzheimer , Morfinanos , NF-kappa B , Camundongos , Animais , NF-kappa B/metabolismo , Transdução de Sinais , Receptor 4 Toll-Like/metabolismo , Neuroimunomodulação , Escopolamina/farmacologia , Inflamação/patologia , Homeostase , Encéfalo/metabolismo , Colinérgicos/farmacologia
3.
Acta Pharmacol Sin ; 44(12): 2504-2524, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37482570

RESUMO

Sinomenine (SIN) is an isoquinoline alkaloid isolated from Sinomenii Caulis, a traditional Chinese medicine used to treat rheumatoid arthritis (RA). Clinical trials have shown that SIN has comparable efficacy to methotrexate in treating patients with RA but with fewer adverse effects. In this study, we explored the anti-inflammatory effects and therapeutic targets of SIN in LPS-induced RAW264.7 cells and in collagen-induced arthritis (CIA) mice. LPS-induced RAW264.7 cells were pretreated with SIN (160, 320, 640 µM); and CIA mice were administered SIN (25, 50 and 100 mg·kg-1·d-1, i.p.) for 30 days. We first conducted a solvent-induced protein precipitation (SIP) assay in LPS-stimulated RAW264.7 cells and found positive evidence for the direct binding of SIN to guanylate-binding protein 5 (GBP5), which was supported by molecular simulation docking, proteomics, and binding affinity assays (KD = 3.486 µM). More importantly, SIN treatment markedly decreased the expression levels of proteins involved in the GBP5/P2X7R-NLRP3 pathways in both LPS-induced RAW264.7 cells and the paw tissue of CIA mice. Moreover, the levels of IL-1ß, IL-18, IL-6, and TNF-α in both the supernatant of inflammatory cells and the serum of CIA mice were significantly reduced. This study illustrates a novel anti-inflammatory mechanism of SIN; SIN suppresses the activity of NLRP3-related pathways by competitively binding GBP5 and downregulating P2X7R protein expression, which ultimately contributes to the reduction of IL-1ß and IL-18 production. The binding specificity of SIN to GBP5 and its inhibitory effect on GBP5 activity suggest that SIN has great potential as a specific GBP5 antagonist.


Assuntos
Artrite Experimental , Artrite Reumatoide , Humanos , Camundongos , Animais , Artrite Experimental/induzido quimicamente , Artrite Experimental/tratamento farmacológico , Interleucina-18/efeitos adversos , Receptores Purinérgicos P2X7/uso terapêutico , Proteína 3 que Contém Domínio de Pirina da Família NLR , Lipopolissacarídeos/farmacologia , Transdução de Sinais , Artrite Reumatoide/tratamento farmacológico , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Proteínas de Ligação ao GTP
4.
Phytomedicine ; 100: 154050, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35397284

RESUMO

BACKGROUND: Sinomenine (SIN) is an anti-inflammatory drug that has been used for decades in China to treat arthritis. In a previous study, SIN acted on α7 nicotinic acetylcholine receptor (α7nAChR) to inhibit inflammatory responses in macrophages, which indicates a new anti-inflammatory mechanism of SIN. However, the level of α7nAChR was increased in the inflammatory responses and was downregulated by SIN in vitro, so the underlying mechanisms of SIN acting on α7nAChR remain unclear. PURPOSE: To analyze the role of α7nAChR in inflammation and the effect and mechanism of SIN regulation of α7nAChR. METHODS: The effects of SIN on α7nAChR in endotoxemic mice and LPS-stimulated macrophages were observed. Nicotine (Nic) was used as a positive control, and berberine (Ber) was used as a negative control targeting α7nAChR. The antagonists of α7nAChR, α-bungarotoxin (BTX) and mecamylamine (Me), were used to block α7nAChR. In RAW264.7 macrophage cells in vitro, α7nAChR short hairpin RNA (shRNA) was used to knock down α7nAChR. Macrophage polarization was analyzed by the detection of TNF-α, IL-6, iNOS, IL-10, Arg-1, and Fizz1. U0126 was used to block ERK phosphorylation. The cytokines α7nAChR, ERK1/2, p-ERK1/2 and Egr-1 were detected. RESULTS: SIN decreased the levels of TNF-α, IL-6 and the expression of α7nAChR increased by LPS in endotoxemic mice. The above effects of SIN were attenuated by BTX. In the α7nAChR shRNA transfected RAW264.7 cells, compared with the control, α7nAChR was knocked down, and M1 phenotype markers (including TNF-α, IL-6, and iNOS) were significantly downregulated, whereas M2 phenotype markers (including IL-10, Arg-1, and Fizz1) were significantly upregulated when stimulated by LPS. SIN inhibited the expression of p-ERK1/2 and the transcription factor Egr-1 induced by LPS in RAW264.7 cells, and the above effects of SIN were attenuated by BTX. The expression of α7nAChR was suppressed by U0126, which lessened the expression of p-ERK1/2 and Egr-1. CONCLUSIONS: SIN acts on α7nAChR to inhibit inflammatory responses and downregulates high expression of α7nAChR in vivo and in vitro. The increase of α7nAChR expression is correlated with inflammatory responses and participates in macrophage M1 polarization. SIN downregulates α7nAChR via a feedback pathway of α7nAChR/ERK/Egr-1, which contributes to inhibiting macrophage M1 polarization and inflammatory responses.


Assuntos
Interleucina-10 , Receptor Nicotínico de Acetilcolina alfa7 , Animais , Anti-Inflamatórios/metabolismo , Anti-Inflamatórios/farmacologia , Retroalimentação , Interleucina-10/metabolismo , Interleucina-6/metabolismo , Lipopolissacarídeos/farmacologia , Macrófagos , Camundongos , Morfinanos , RNA Interferente Pequeno/farmacologia , Fator de Necrose Tumoral alfa/metabolismo , Receptor Nicotínico de Acetilcolina alfa7/metabolismo
5.
J Leukoc Biol ; 110(6): 1113-1120, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34425026

RESUMO

Sinomenine (SIN) is a clinical drug for treating rheumatoid arthritis (RA) in China. Our previous study found SIN inhibited inflammation via alpha7 nicotinic acetylcholine receptor (α7nAChR) in macrophages in vitro. Adenosine receptor A2A has anti-inflammatory and immunosuppressive function. However, the mechanisms of SIN acting on α7nAChR and the effect on adenosine A2A receptor (A2A R) in RA are not clear. In the present study, the effects of SIN on adjuvant-induced-arthritis (AIA) rats in vivo and on fibroblast-like synoviocytes (FLSs) in vitro were investigated. Indomethacin (Indo) and methotrexate (MTX), the clinical anti-arthritis drugs, were used as controls. Nicotine (Nic), a specific agonist of α7nAChR, was used as a control for targeting α7nAChR. Alpha-bungarotoxin (α-BTX), the antagonist of α7nAChR or small interference RNA (siRNA) was used to block or knock down α7nAChR. Results showed that SIN decreased arthritis index, hind paw volume, erythrocyte sedimentation (ESR) and serum TNF-α in AIA rats, and α-BTX attenuated the earlier-mentioned effects of SIN and Nic, but not Indo and MTX. The expressions of A2A R in synovium declined in AIA rats, but remarkably increased after the intervention of SIN. The expression of A2A R decreased by LPS or TNF-α, but increased by SIN; cAMP also increased by SIN in FLSs in vitro. SIN inhibited the expression of MCP-1, IL-6, and vascular endothelial growth factor in LPS-induced FLSs. SIN inhibited the activation of NF-κB. Meanwhile, α-BTX or α7nAChR siRNA blocked the earlier-mentioned effects of SIN in FLSs. Results suggested the expressions of A2A R in synovium and FLSs are negatively correlated with the arthritis progression of AIA rats and the activation of FLSs. SIN increases A2A R and inhibits the activation of NF-κB pathway via α7nAChR in AIA rats and FLSs.


Assuntos
Antirreumáticos/farmacologia , Artrite Reumatoide/metabolismo , Morfinanos/farmacologia , NF-kappa B/metabolismo , Receptor A2A de Adenosina/metabolismo , Receptor Nicotínico de Acetilcolina alfa7/metabolismo , Animais , Artrite Experimental/metabolismo , Artrite Experimental/patologia , Artrite Reumatoide/patologia , Masculino , Ratos , Ratos Sprague-Dawley , Sinoviócitos/metabolismo
6.
J Leukoc Biol ; 109(4): 843-852, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-32726882

RESUMO

Lung cancer is the leading cause of cancer deaths worldwide, with a high morbidity and less than 20% survival rate. Therefore, new treatment strategies and drugs are needed to reduce the mortality of patients with lung cancer. α7 nicotinic acetylcholine receptor (α7 nAChR), as a receptor of nicotine and its metabolites, is a potential target for lung cancer treatment. Our previous studies revealed that sinomenine plays anti-inflammation roles via α7 nAChR and down-regulates the expression of this receptor, thus increasing the inflammatory response. Hence, sinomenine is possibly a natural ligand of this receptor. In the present study, the effects of sinomenine on lung cancer A549 cells and tumor-bearing mice were determined to investigate whether this alkaloid has an inhibitory effect on lung cancer via α7 nAChR. CCK-8 assay, wound-healing test, and flow cytometry were performed for cell proliferation, cell migration, and apoptosis analysis in vitro, respectively. Xenograft mice were used to evaluate the effects of sinomenine in vivo. Results showed that sinomenine decreased cell proliferation and migration abilities but increased the percentage of apoptotic cells. Tumor volume in tumor-bearing mice was significantly reduced after sinomenine treatment compared with that in the vehicle group mice (p < 0.05). Furthermore, the effects of sinomenine were abolished by the α7 nAChR antagonist mecamylamine and the allosteric modulator PNU-120596, but no change occurred when the mice were pretreated with the muscarinic acetylcholine receptor antagonist atropine. Meanwhile, sinomenine suppressed α7 nAChR expression in vitro and in vivo, as well as the related signaling molecules pERK1/2 and ERK1/2 and the transcription factors TTF-1 and SP-1. By contrast, sinomenine up-regulated the expression of another transcription factor, Egr-1. These effects were restricted by mecamylamine and PNU but not by atropine. Results suggested that sinomenine can inhibit lung cancer via α7 nAChR in a negative feedback mode.


Assuntos
Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Morfinanos/farmacologia , Receptor Nicotínico de Acetilcolina alfa7/metabolismo , Células A549 , Animais , Antineoplásicos/farmacologia , Humanos , Masculino , Camundongos Endogâmicos C57BL , Transdução de Sinais/efeitos dos fármacos , Fatores de Transcrição/metabolismo
7.
Artigo em Inglês | MEDLINE | ID: mdl-31186658

RESUMO

Sinomenine (SIN) is the active ingredient of the Chinese herb Sinomenium acutum that has been used to treat rheumatoid arthritis (RA) for about 30 years in China. Marked expression of the alpha7 nicotinic acetylcholine receptor (α7nAChR) in the joint synovium of RA patients suggested a relationship between α7nAChR and RA. This study investigated the relationship between α7nAChR and RA development and the effects of SIN on α7nAChR expression in vivo and in vitro. Sprague-Dawley rats were injected with complete Freund's adjuvant to induce arthritis and then treated with SIN or methotrexate (MTX) from day 0 to day 30. Four clinical parameters-paw volume, arthritic index (AI), serum TNF-α concentration, and erythrocyte sedimentation rate (ESR)-were measured. Splenic lymphocytes were isolated for Bacille Calmette Guerin (BCG) stimulation. α7nAChR expression in tissues and cells was examined by RT-PCR, western blot, immunofluorescence, flow cytometry, and immunohistochemistry. Cell proliferation was evaluated by the CCK-8 assay. The relationship between α7nAChR expression and the four clinical parameters was analyzed by single-factor correlation analysis. Our results showed that the paw volume, AI, TNF-α concentration, and ESR in adjuvant-induced arthritic (AIA) rats were reduced by SIN or MTX treatment. SIN decreased α7nAChR expression in tissues and cells compared to the model group, while MTX had no significant effect on α7nAChR expression. Moreover, there was a positive relationship between α7nAChR expression and paw swelling, AI, and TNF-α concentration. Splenic lymphocyte activation was accompanied by increased α7nAChR expression, while SIN treatment inhibited cell activation and downregulated α7nAChR expression. α7nAChR expression showed a positive correlation with the progression of RA in AIA rats that may involve lymphocyte activation. Different from MTX, the inhibition of SIN on α7nAChR expression might contribute to its antiarthritic effect, suggesting that SIN could be an important supplement to the treatment strategy for RA.

8.
Immunopharmacol Immunotoxicol ; 41(1): 172-177, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30896303

RESUMO

Objective: To investigate the cellular mechanism that sinomenine (SIN) inhibits inflammation in macrophages induced by LPS through α7 nicotinic acetylcholine receptor (α7nAChR). Materials and methods: RAW264.7 cells were stimulated with LPS and treated by SIN or nicotine (Nic). A selective antagonist of α7nAChR, α-bungarotoxin (BTX) was used to block α7nAChR. AG490 was used to inhibit JAK2 activation. ELISA was performed to detect the levels of TNF-α and MCP-1. Western blotting was used to analyze the expression of MIF, MMP-9, CD14, TLR4, STAT3 and p-STAT3. Intracellular-free calcium level was measured by Fluorescent probe fluo-3/AM Results: SIN inhibited the production of TNF-α, MCP-1, MIF, and MMP-9, decreased the expression of CD14 and TLR4, and inhibited the release of intracellular-free calcium from intracellular stores in RAW 264.7 cells stimulated by LPS. JAK-specific inhibitor AG490 attenuated the inhibitory effect of SIN on TNF-α. SIN increased the phosphorylation of STAT3. And the above effects of SIN were attenuated by antagonist of α7nAChR. Conclusions: SIN can decrease the expression of CD14/TLR4 and intracellular free calcium level, activate JAK2/STAT3 pathway to inhibit inflammatory response through α7nAChR in macrophages.


Assuntos
Anti-Inflamatórios/farmacologia , Cálcio/metabolismo , Janus Quinase 2/metabolismo , Receptores de Lipopolissacarídeos/metabolismo , Morfinanos/farmacologia , Fator de Transcrição STAT3/metabolismo , Receptor 4 Toll-Like/metabolismo , Receptor Nicotínico de Acetilcolina alfa7/metabolismo , Animais , Lipopolissacarídeos/toxicidade , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/metabolismo , Camundongos , Células RAW 264.7 , Transdução de Sinais
10.
Int Immunopharmacol ; 56: 65-70, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29367088

RESUMO

Fibroblast like synoviocyte (FLS) is a crucial in the pathogenesis of rheumatoid arthritis (RA), and involved in inflammation and joint destruction. Sinomenine (SIN), an alkaloid derived from the plant Sinomenium acutum, has anti-inflammatory and analgesic effect and been used for RA treatment in China. Alpha 7 nicotinic acetylcholine receptors (α7nAChR), as the key receptor in cholinergic anti-inflammatory pathway (CAP) to inhibit inflammation, has been detected in RA patients synovium, but its role is still unclear. Here we investigated the association between the aggressive proliferation of FLS and α7nAChR expression and the effect of sinomenine. FLS was isolated from synovial tissues of adjuvant-induced-arthritis (AIA) rat. Tumor necrosis factor(TNF)-α was used to induce the aggressive proliferation of FLS. MTT assay was applied to evaluate the proliferation of FLS. The messenger RNA (mRNA) and protein levels of α7nAChR and early growth response gene-1 (Egr-1) were measured. The results showed that TNF-α induced FLS proliferation in vitro (P < .01) and increased the phosphorylation of ERK1/2 and the expression of Egr-1 and α7nAChR (P < .05 or P < .01). U0126, the inhibitor of ERK1/2 inhibited α7nAChR expression and FLS proliferation significantly (P < .05 or P < .01). Specific short interference RNA(siRNA) of α7nAChR decreased α7nAChR expression and inhibited FLS proliferation as well. SIN inhibited the proliferation of FLS and decreased the phosphorylation of ERK1/2, and the expression of Egr-1 and α7nAChR induced by TNF-α (P < .05). In conclusion, the expression of α7nAChR involved in the aggressive proliferation of FLS induced by TNF-α and was regulated by ERK/Egr-1 signal pathway. SIN inhibited FLS proliferation and α7nAChR expression through inhibiting ERK/Egr-1 signal pathway, this may contribute to the anti-inflammatory and anti-arthritic effect of SIN.


Assuntos
Anti-Inflamatórios/uso terapêutico , Artrite Experimental/tratamento farmacológico , Artrite Reumatoide/tratamento farmacológico , Morfinanos/uso terapêutico , Sinoviócitos/imunologia , Receptor Nicotínico de Acetilcolina alfa7/metabolismo , Animais , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Modelos Animais de Doenças , Proteína 1 de Resposta de Crescimento Precoce/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Regulação da Expressão Gênica , Humanos , Masculino , RNA Interferente Pequeno/genética , Ratos , Ratos Sprague-Dawley , Transdução de Sinais , Sinomenium/imunologia , Sinoviócitos/efeitos dos fármacos , Receptor Nicotínico de Acetilcolina alfa7/genética
11.
Pharmacol Res ; 125(Pt B): 150-160, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-28867637

RESUMO

Zhengqing Fengtongning (ZQFTN), the pharmaceutical preparation of sinomenine (SIN) derived from the medicinal plant Sinmenium acutum, is well-known in China as an effective treatment for rheumatoid arthritis (RA). However, its histamine-release anaphylactoid reactions (HRARs) occur often in some patients. Therefore, it is desirable to establish effective clinical protocols to manage such HRARs. In the study, rat models with systemic HRARs and local HRARs of the skin were established. The level of vascular permeability and mast cell numbers was determined by quantitative analysis using Evans blue dye and histological assays. The levels of histamine, leukotriene B4 (LTB4) and IL-33 in plasma were detected by UHPLC-SPE-MS, ELISA and immunohistochemistry assays, respectively. The results demonstrated that SIN significantly induced both systemic and local HRARs in rats, showing significant decrease of body temperature, increases in vascular permeability in skin, injury of lung tissues and mast cell infiltration and IL-33 expression in skin and lung tissues. Mechanistic study showed that tranilast could prevent SIN-triggered HRARs via inhibition of H1 receptor gene expression and NF-κB signaling. Our findings provide evidence that mast cell membrane stabilizers and H1 receptor blockers effectively prevent SIN-induced HRARs, and cromolyn, cetirizine and tranilast can be used in the clinic for the management of HRARs induced by ZQFTN.


Assuntos
Anafilaxia/tratamento farmacológico , Antialérgicos/uso terapêutico , Antagonistas dos Receptores Histamínicos H1/uso terapêutico , Morfinanos , NF-kappa B/antagonistas & inibidores , ortoaminobenzoatos/uso terapêutico , Lesão Pulmonar Aguda/sangue , Lesão Pulmonar Aguda/induzido quimicamente , Lesão Pulmonar Aguda/imunologia , Anafilaxia/sangue , Anafilaxia/induzido quimicamente , Anafilaxia/imunologia , Animais , Antialérgicos/farmacologia , Cetirizina/farmacologia , Cetirizina/uso terapêutico , Cromolina Sódica/farmacologia , Cromolina Sódica/uso terapêutico , Feminino , Histamina/sangue , Antagonistas dos Receptores Histamínicos H1/farmacologia , Liberação de Histamina/efeitos dos fármacos , Interleucina-33/imunologia , Leucotrieno B4/sangue , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Mastócitos/efeitos dos fármacos , Mastócitos/imunologia , NF-kappa B/imunologia , Ratos Sprague-Dawley , Receptores Histamínicos H1/imunologia , Transdução de Sinais/efeitos dos fármacos , Pele/efeitos dos fármacos , Pele/imunologia , ortoaminobenzoatos/farmacologia
12.
Biochem Pharmacol ; 142: 133-144, 2017 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-28711625

RESUMO

Recently, microsomal prostaglandin E synthase 1 (mPGES-1) has attracted much attention from pharmacologists as a promising strategy and an attractive target for treating various types of diseases including rheumatoid arthritis (RA), which could preserve the anti-inflammatory effect while reducing the adverse effects often occur during administration of non-steroidal anti-inflammatory drugs (NSAIDs). Here, we report that sinomenine (SIN) decreased prostaglandin (PG)E2 levels without affecting prostacyclin (PG)I2 and thromboxane (TX)A2 synthesis via selective inhibiting mPGES-1 expression, a possible reason of low risk of cardiovascular event compared with NSAIDs. In addition, mPGES-1 protein expression was down-regulated by SIN treatment in the inflamed paw tissues both in carrageenan-induced edema model in rats and the collagen-II induced arthritis (CIA) model in DBA mice. More interestingly, SIN suppressed the last step of mPGES-1 gene expression by decreasing the DNA binding ability of NF-κB, paving a new way for drug discovery.


Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Artrite Experimental/tratamento farmacológico , Edema/tratamento farmacológico , Expressão Gênica/efeitos dos fármacos , Morfinanos/uso terapêutico , Prostaglandina-E Sintases/genética , Células A549 , Animais , Anti-Inflamatórios não Esteroides/efeitos adversos , Anti-Inflamatórios não Esteroides/isolamento & purificação , Anti-Inflamatórios não Esteroides/farmacologia , Artrite Experimental/imunologia , Técnicas de Cultura de Células , Sobrevivência Celular/efeitos dos fármacos , Edema/imunologia , Feminino , Macrófagos Peritoneais/efeitos dos fármacos , Masculino , Camundongos Endogâmicos DBA , Morfinanos/efeitos adversos , Morfinanos/isolamento & purificação , Morfinanos/farmacologia , Ratos Sprague-Dawley , Transfecção
13.
Pharmacol Res ; 111: 303-315, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27343699

RESUMO

Inflammation is a defensive reaction of body to resist foreign invasion. However, it has been demonstrated that excessive and continuous inflammatory responses contribute to various inflammatory diseases, including rheumatoid arthritis. Nuclear factor-κB (NF-κB) regulates the expression of an array of inflammatory mediators, cytokines and chemokine genes in activated macrophages. Therefore, NF-κB has become an attractive drug target for controlling inflammation. In this study, stauntoside B, a C21 steroidal glycosides compound isolated from a Chinese medicine Cynanchi Stauntonii, was for the first time found to suppress macrophage activation induced by lipopolysaccharide (LPS) in RAW264.7 cells and rat primary peritoneal macrophages and could be a potent NF-κB inhibitor. The results showed that stauntoside B significantly reduced the release of inflammatory mediators in activated RAW264.7 cells and rat peritoneal macrophages, including nitric oxide (NO) and prostaglandin E2 (PGE2). The mRNA expressions of pro-inflammatory mediators and cytokines, including inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), microsomal prostaglandin synthetase-1 (mPGES-1), tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), interleukin-6 (IL-6), and monocyte chemoattractant protein-1 (MCP-1) as well as the production of TNF-α and IL-6 were also inhibited by stauntoside B. Mechanistic investigation implies that the anti-inflammatory activity of stauntoside B could result from the suppression of LPS-induced IKKα/ß activation, IκBα phosphorylation, p65 (ser536) NF-κB phosphorylation, and ERK MAPK activation by stauntoside B treatment in activated macrophages. Meanwhile, stauntoside B could induce apoptosis in LPS-activated macrophages. The current study suggests stauntoside B being a valuable candidate drug for the treatment of inflammatory diseases, especially for NF-κB activation associated inflammatory diseases.


Assuntos
Anti-Inflamatórios/farmacologia , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Inflamação/prevenção & controle , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos Peritoneais/efeitos dos fármacos , NF-kappa B/metabolismo , Saponinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Esteroides/farmacologia , Animais , Apoptose/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ativação Enzimática , Inflamação/enzimologia , Inflamação/genética , Mediadores da Inflamação/metabolismo , Lipopolissacarídeos/farmacologia , Macrófagos Peritoneais/enzimologia , Camundongos , Fosforilação , Células RAW 264.7 , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos Sprague-Dawley
14.
Shock ; 44(2): 188-95, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25895149

RESUMO

Sinomenine (SIN), an alkaloid derived from the plant Sinomenium acutum, has anti-inflammatory and analgesic effects and has been used for rheumatoid arthritis treatment in China. This study aims to verify the hypothesis that SIN acts on α7 nicotinic acetylcholine receptor (α7nAChR) to inhibit the activation of macrophages stimulated by lipopolysaccharide. The prototypical α7nAChR antagonist α-bungarotoxin and mecamylamine attenuated the effect of SIN on tumor necrosis factor-α and interleukin-6 in RAW264.7 murine macrophage-like cells and primary peritoneal macrophages of mouse induced by lipopolysaccharide. With the knockdown of α7nAChR expression in RAW264.7 cells by small interfering RNA, the inhibitory effect of SIN on tumor necrosis factor-α and interleukin-6 was reversed. Sinomenine decreased p65 expression in nuclear and increased IκBα expression in cytoplasm, and these effects were reversed by the α7nAChR small interfering RNA as well. These results indicate that the anti-inflammatory effects of SIN on macrophages in vitro depend on α7nAChR.


Assuntos
Anti-Inflamatórios/química , Lipopolissacarídeos/química , Macrófagos Peritoneais/efeitos dos fármacos , Morfinanos/química , Receptor Nicotínico de Acetilcolina alfa7/metabolismo , Animais , Bungarotoxinas/química , Citoplasma/metabolismo , Proteínas I-kappa B/metabolismo , Interleucina-6/metabolismo , Macrófagos Peritoneais/metabolismo , Masculino , Mecamilamina/química , Camundongos , Camundongos Endogâmicos BALB C , Inibidor de NF-kappaB alfa , Células RAW 264.7 , Interferência de RNA , RNA Interferente Pequeno/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
15.
J Cardiovasc Pharmacol ; 63(4): 323-32, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24710469

RESUMO

The adherence to circulating leukocytes, such as monocytes and neutrophils, to vascular endothelial cells is of central importance to the pathogenesis of various cardiovascular diseases (CVDs) including atherosclerosis and myocardial ischemia-reperfusion injury. Compound danshen dripping pill (CDDP; Fufang Danshen Diwan in Chinese), namely cardiotonic pill, is extensively used for CVDs medication in China and some other countries. Here, we sought to investigate the effect of CDDP on leukocytes binding to vascular endothelial cells and elaborate the possibly underlying mechanism. Using seropharmacological method, rabbit sera containing CDDP were shown to mitigate the adhesiveness of monocytes and neutrophils to tumor necrosis factor alpha-stimulated human umbilical vein endothelial cells in dose and time-dependent manners, alleviate the levels of intercellular adhesion molecule 1 and vascular cell adhesion molecule 1 messenger RNA and protein dose dependently and also encumber IκBα degradation, p65 nuclear translocation, nuclear factor-kappaB (NF-κB) DNA-binding activity, and NF-κB-responsive gene transcription in tumor necrosis factor alpha-activated human umbilical vein endothelial cells. This study suggests that CDDP protects against CVDs potentially by attenuation of leukocytes-endothelium adhesion cascade via lessening endothelial cell adhesion molecules expression and NF-κB signaling pathway activity.


Assuntos
Adesão Celular/efeitos dos fármacos , Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos , Molécula 1 de Adesão Intercelular/biossíntese , Leucócitos/efeitos dos fármacos , NF-kappa B/efeitos dos fármacos , Salvia miltiorrhiza/química , Fator de Necrose Tumoral alfa/farmacologia , Molécula 1 de Adesão de Célula Vascular/biossíntese , Animais , Células Endoteliais/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Monócitos/efeitos dos fármacos , Neutrófilos/efeitos dos fármacos , Coelhos , Transdução de Sinais/efeitos dos fármacos
16.
Zhongguo Zhong Yao Za Zhi ; 35(13): 1660-5, 2010 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-20862951

RESUMO

OBJECTIVE: To transform the antimicrobial peptide fusion gene of cecropin B and rabbit NP-1(CN) into Houttuynia cordata to improve its antimicrobic capability. METHOD: The fusion gene of CN designed and synthesized artificially was recombined with expression vector pBI121. The recombined vector was transformed to Agrobacterium tumefaciens LBA4404, by which CN gene was transformed to the explants of H. cordata. The transgenic regeneration plantlets were selected by kanamycin and rapid screening PCR. The transgenic plants were identified by PCR-Southern of genomic DNA and RT-PCR. The disease resistances were detected by antibacterial zone trail of leaf extracts to E. coli K12 and infection by Rhizoctonia solani. RESULT: Gene of interesting CN was inserted into genomic DNA and expressed in transformed H, cordata, whose resistance to E. coli K12 and Rh. solani was stronger than that of the non-transformed control. CONCLUSION: The fusion gene CN can improve antimicrobic capability of transformed H. cordata.


Assuntos
Proteína C-Reativa/genética , Houttuynia/genética , Proteínas de Insetos/genética , Proteínas do Tecido Nervoso/genética , Plantas Geneticamente Modificadas/genética , Transformação Genética , Animais , Antibacterianos/imunologia , Antibacterianos/farmacologia , Proteína C-Reativa/metabolismo , Proteína C-Reativa/farmacologia , Houttuynia/imunologia , Houttuynia/microbiologia , Imunidade Inata , Proteínas de Insetos/imunologia , Proteínas de Insetos/farmacologia , Proteínas do Tecido Nervoso/metabolismo , Proteínas do Tecido Nervoso/farmacologia , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Plantas Geneticamente Modificadas/imunologia , Plantas Geneticamente Modificadas/microbiologia , Coelhos , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/farmacologia , Rhizoctonia/fisiologia
17.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi ; 20(1): 113-6, 2004 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-15182636

RESUMO

AIM: To study the effects of total alkaloid of Tongbiling prescription(TBL) on Th1 type cytokine expression in T cells in order to elucidate the anti-inflammatory mechanism of TBL. METHODS: The lymphocytes were isolated from mouse mesenteric lymph nodes and cultured in-vitro. Various concentrations of TBL were added to the culture followed by phorbol ester and inomycin treatment and then incubated for another 4 hours. The expressions of IFN-gamma and TNF-alpha in the lymphocytes were analyzed by flow cytometry. RESULTS: 200 mg/L and 100 mg/L TBL could obviously inhibit IFN-gamma and TNF-alpha expressions in T lymphocytes. CONCLUSION: Inhibiting Th1 cytokine expression may be one mechanism by which TBL can treat rheumatoid arthritis.


Assuntos
Alcaloides/farmacologia , Medicamentos de Ervas Chinesas/farmacologia , Interferon gama/análise , Linfócitos T/efeitos dos fármacos , Fator de Necrose Tumoral alfa/análise , Animais , Artrite Reumatoide/tratamento farmacológico , Artrite Reumatoide/imunologia , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Linfócitos T/imunologia , Células Th1/imunologia
18.
Artigo em Chinês | MEDLINE | ID: mdl-14764218

RESUMO

OBJECTIVE: To investigate the gene expression of some lipopolysaccharide (LPS) receptors after LPS stimulation. METHODS: The total RNA from normal and LPS-challenged mice was extracted by Trizol reagent and the gene expression of Toll-like receptor 2 (TLR2), TLR4, CD(14), LPS-binding protein (LBP) and tumor necrosis factor-alpha(TNF-alpha) were measured by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). RESULTS: The gene of TLR2 was expressed in normal lungs and spleen tissues, and TLR2, CD(14), LBP in liver. After challenged by LPS, the expressions of TLR2, TLR4, and TNF-alpha in lungs, TLR2, CD(14), LBP and TNF-alpha in liver, TLR2, TLR4, CD(14), and TNF-alpha in spleen were increased at 1, 3, and 5 hours. CONCLUSION: LPS might alert the ability against pathogen-associated molecules by inducing or enhancing the expression of genes that involved in the LPS signal transduction.


Assuntos
Proteínas de Fase Aguda , Glicoproteínas de Membrana/genética , Proteínas Monoméricas de Ligação ao GTP/farmacologia , Receptores de Superfície Celular/genética , Animais , Proteínas de Transporte/genética , Expressão Gênica/efeitos dos fármacos , Fígado/metabolismo , Pulmão/metabolismo , Masculino , Camundongos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Baço/metabolismo , Receptor 2 Toll-Like , Receptor 4 Toll-Like , Receptores Toll-Like , Fator de Necrose Tumoral alfa/genética , Proteína ran de Ligação ao GTP
19.
Zhongguo Zhong Yao Za Zhi ; 28(4): 352-5, 2003 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-15139149

RESUMO

OBJECTIVE: To observe in vitro the effect of Sinomenine, a pure alkaloid extracted from the chinese medical plant Sinomenium acutum on the activity of cyclooxygenase (COX-1 and COX-2) and the expression of COX-1 and COX-2 mRNA. METHOD: Mononuclear leukocytes were obtained from healthy adults. Isolated mononuclear leucocytes from human peripheral blood (PBMC) were incubated (1 x 10(6).mL-1) with or without sinomenine (or indomethacin), after incubated for 24 hours at 37 degrees C with 5% CO2; the media were assayed for the PGE2 by radioimmunoassay (RIA). LPS was used to stimulate the monocytes at a concentration of 5 micrograms.mL-1. And by RT-PCR, both COX-1 and COX-2 mRNAs were detected in Mononuclear leukocytes after incubation for different hours with drug (sinomenine or indomethacin) or not. RESULT: LPS (stimulated) induced the production of PGE2 in PBMC increasing with high expression of COX-2 mRNA; sinomenine reduced PGE2 production in LPS stimulated human monocytes more than in non-stimulated human monocytes. In comparative experiments, indomethacin, a non selective COX inhibitor, reduced the production of PGE2 equally in both states. Meanwhile, neither sinomenine(0.1-1 mmol.L-1) nor indomethacin(0.5-10 mumol.L-1) inhibited the expression of both COX-1 and COX-2 mRNAs by RT-PCR with beta-actin as reference. CONCLUSION: In contrast with indomethacin, Sinomenine shows a preferential inhibitory effect on COX-2 over COX-1, These results suggest that Sinomenine is a selective COX-2 inhibitor, which may be directly related to suppressing cyclooxygenase activity.


Assuntos
Isoenzimas/biossíntese , Leucócitos Mononucleares/enzimologia , Morfinanos/farmacologia , Prostaglandina-Endoperóxido Sintases/biossíntese , Prostaglandina-Endoperóxido Sintases/metabolismo , Adulto , Ciclo-Oxigenase 1 , Ciclo-Oxigenase 2 , Dinoprostona/sangue , Humanos , Proteínas de Membrana , Morfinanos/isolamento & purificação , Plantas Medicinais/química , RNA Mensageiro/biossíntese , Sinomenium/química
20.
Zhong Yao Cai ; 26(10): 751-3, 2003 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-14768397

RESUMO

OBJECTIVE: To supply basic research materials for clarifying the immunocompetence materials of Sijunzi Decoction. METHODS: Two polysaccharides(SJZPS-Vb-1-2) were purified with Sephadex DEAE A 25 and Sephadex G 200 from SJZPS-Vb part (with the highest immunocompetence). Their molecular weights were determined. The sugar compositions were analyzed with GC. The linkage positions of the component sugars were determined by methylation and GC/MS. RESULTS: SJZPS-Vb-1-2 was uonic-containing heteropolysaccharides with the component sugars of glucose, galactose and mannose in different molar ratios. The molecular weight of SJZPS-VB-1 was 38,300, SJZPS-VB-2, 26,000. The molar ratio of SJZPS-Vb-1 was as glu:gal:man 1:046:2.15 and SJZPS-Vb-2, glu:gal:man 1:1.41:4.18.


Assuntos
Medicamentos de Ervas Chinesas/química , Proteínas Hemolisinas/imunologia , Monossacarídeos/análise , Plantas Medicinais/química , Polissacarídeos/isolamento & purificação , Animais , Combinação de Medicamentos , Medicamentos de Ervas Chinesas/isolamento & purificação , Feminino , Galactose/análise , Glucose/análise , Imunocompetência/imunologia , Masculino , Manose/análise , Camundongos , Peso Molecular , Polissacarídeos/química , Polissacarídeos/imunologia
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