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Objective To explore the effect of the combination of Tuina and treadmill training on nerve regeneration after sciatic nerve transection.Methods A total of 60 Sprague-Dawley rats were randomly divided into control group,model group and treatment group equal-ly.The sciatic nerve was transected in the latter two groups and the epineurium suture was delayed four days.Then the treatment group ac-cepted Tuina and treadmill training once a day.The motor conduction velocity(MCV)of sciatic nerve was measured one and two months af-ter intervention in ten rats respectively,while the number of regenerated axons and Schwann cells was counted,and structures of sciatic nev-er were observed under electron microscopy. Results Compared with the model group, the MCV accelerated in the treatment group (P<0.05),the number of axons was not significantly different(P>0.05),and the number of Schwann cells increased two months after modeling (P<0.05),with less injury under electron microscopy.Conclusion Tuina combined with treadmill training can promote the regeneration of the injured peripheral nerve.
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[This corrects the article DOI: 10.1093/ecam/neq011.].
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Ampelopsis sinica root is widely used in Chinese folk medicine for treating liver disorders caused by the hepatitis B virus (HBV). The present study was performed in order to investigate the anti-HBV activity and mechanisms of the ethanol extract from A. sinica root (EASR) in vitro. The antiviral activity of EASR was examined by detecting the levels of HBsAg, HBeAg and extracellular HBV DNAs in stable HBV-producing human hepatoblastoma HepG2 2.2.15 cells. We found that EASR effectively suppressed the secretion of HBsAg and HBeAg from HepG2 2.2.15 cells in a dose-dependent manner, and it also suppressed the amount of extracellular HBV DNA. After EASR treatment, the percentage of apoptotic cells was found to be significantly higher than that of control by flow cytometric analysis. A luciferase reporter gene assay was used to determine the effects of EASR on the activities of HBV promoters and intracellular signaling pathways. The results showed that EASR selectively inhibited the activities of HBV promoters (Cp, S1p and Fp) and the p53 signaling pathway in HepG2 cells significantly. These data indicate that EASR exerts anti-HBV effects via inhibition of HBV promoters and the p53-associated signaling pathway, which helps to elucidate the mechanism underlying the potential therapeutic value of EASR.
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Despite the availability of an effective vaccine, the hepatitis B virus (HBV) infection and its treatment remains one of the foremost public health problems in the world. The present study was performed in order to investigate the anti-HBV activity of lutein in vitro. The antiviral activity of lutein was examined by detecting the levels of HBsAg, HBeAg and extracellular HBV DNA in stable HBV-producing human hepatoblastoma HepG2 2.2.15 cells. It was found that lutein effectively suppressed the secretion of HBsAg from HepG2 2.2.15 cells in a dose-dependent manner, and it also suppressed the amount of extracellular HBV DNA. A luciferase reporter gene assay was used to determine the effects of lutein on the activities of HBV promoters. The results showed that lutein inhibited the activity of HBV full-length promoter (Fp). These data indicate that lutein possesses an anti-HBV activity and exerts its antivirus effects via inhibition of HBV transcription.
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Antivirais/farmacologia , Vírus da Hepatite B/efeitos dos fármacos , Luteína/farmacologia , DNA Viral/análise , Células Hep G2 , Antígenos de Superfície da Hepatite B/análise , Antígenos E da Hepatite B/análise , Humanos , Regiões Promotoras Genéticas/efeitos dos fármacos , Transcrição Gênica/efeitos dos fármacosRESUMO
Melilotus suaveolens Ledeb is a traditional medicinal plant for treating inflammation-related disease. This explores the inner anti-inflammatory mechanism of n-butanol extract from M. suaveolens Ledeb. Inflammatory cellular model was established by lipopolysaccharide intervention on RAW264.7 cell line. Levels of secreted cytokines TNF-α, IL-1ß, IL-6, NO and IL-10 in supernatant, mRNA expression of TNF-α, COX-2, iNOS and HO-1, protein expression of COX-2 and HO-1, activation of NF-κB and ingredients in the extract were assayed by ELISA, real time quantitative PCR, western blot, immunocytochemical test and HPLC fingerprint test, respectively. As a result, the extract could not only markedly reduce the production of pro-inflammatory mediators to different extents by blocking NF-κB activation but also promote the release of anti-inflammatory mediator HO-1 significantly. Each 1 g extract contained 0.023531 mg coumarin and another two high polar ingredients, probably saponins. It can be concluded that the extract has similar effects on antagonizing pro-inflammatory mediators and cytokines like Dexamethasone, and has effects on promoting the production of anti-inflammatory mediators.
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AIM OF THE STUDY: This paper aimed to elucidate the anti-inflammatory effects of EtOAc fraction prepared from Melilotus suaveolens Ledeb ethanol extract with a cellular model of LPS-stimulated RAW 264.7 cell. MATERIALS AND METHODS: Some key pro-inflammatory cytokines and mediators including IL-1 beta, IL-6, NO, iNOS, COX-2 and TNF-alpha, two important anti-inflammatory cytokines and mediators IL-10 and HO-1, I-kappaB and NF-kappaB were studied by sandwich ELISA, real-time PCR, western blot analysis and immunocytochemistry. At last a HPLC fingerprint was taken to evaluate the fraction. RESULTS: The EtOAc fraction could significantly inhibit the production of IL-1 beta, IL-6, NO, TNF-alpha, COX-2 in LPS-stimulated cell than that of single LPS-stimulated cell (p<0.01 or p<0.05), and the extract could increase the production of IL-10 and HO-1 than that of single LPS intervention cell (p<0.01 or p<0.05). Meanwhile, the extract also could inhibit the production of NF-kappaB compared to single LPS-stimulated cell. All the results showed that the extract had a good anti-inflammatory effect on LPS-stimulated RAW264.7 cell. CONCLUSIONS: Taken together, the anti-inflammatory actions of M. suaveolens Ledeb EtOAc fraction might be due to the down-regulation of IL-1 beta, IL-6, NO, TNF-alpha and COX-2 via the suppression of NF-kappaB activation, and another pathway was up regulating the production of IL-10 and HO-1. Meanwhile, the EtOAc fraction might be further studied to isolate the active anti-inflammatory ingredients besides coumarin.
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Acetatos/química , Anti-Inflamatórios/uso terapêutico , Lipopolissacarídeos/farmacologia , Melilotus/química , Extratos Vegetais/uso terapêutico , Animais , Sequência de Bases , Linhagem Celular , Cromatografia Líquida de Alta Pressão , Citocinas/análise , Primers do DNA , Ensaio de Imunoadsorção Enzimática , Mediadores da Inflamação/análise , Ativação de Macrófagos , Macrófagos/efeitos dos fármacos , Camundongos , Reação em Cadeia da PolimeraseRESUMO
AIM: To study the anti-inflammatory effect of petroleum ether extract from Melilotus suaveolens Ledeb. METHODS: Inflammatory cell model was constructed by LPS acting on the RAW264.7 cell line. The expression and distribution of NF-kappaB were detected using immunocytochemical method. The expression of mRNA and protein of Heme oxygenase 1(HO-1) were detected by real-time PCR and Western blot. RESULTS: The immunocytochemical analysis showed that the cytoplasm stained to brown presented NF-kappaB inactivation after the intervention of petroleum ether extract while the cell nucleus stained to brown presented NF-kappaB activation after the only intervention of LPS. The expression of HO-1 mRNA was significantly enhanced by the extract in a dose-dependent manner, and the expression of HO-1 protein was markedly enhanced too. CONCLUSION: The petroleum ether extract from Melilotus suaveolens Ledeb can resist inflammation by inhibiting the activation of proinflammatory factor NF-kappaB and enhancing the expression of anti-inflammatory factor HO-1.
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Anti-Inflamatórios/farmacologia , Expressão Gênica/efeitos dos fármacos , Heme Oxigenase-1/genética , Melilotus/química , NF-kappa B/genética , Extratos Vegetais/farmacologia , Animais , Linhagem Celular Tumoral , Éter/química , Heme Oxigenase-1/imunologia , Inflamação/tratamento farmacológico , Inflamação/genética , Inflamação/imunologia , Camundongos , NF-kappa B/imunologiaRESUMO
This study is to explore the anti-inflammatory mechanism of the ethanol extract of Duchesnea indica (Andr) Focke. An inflammatory cellular model was established by addition of lipopolysaccharide (LPS) on RAW264.7 cell line. The cellular secretion of TNF-alpha, IL-1beta, IL-6, NO and IL-10 in supernatant, mRNA expression of TNF-alpha, COX-2, iNOS and HO-1, protein expression of COX-2 and HO-1, and activation of NF-kappaB were assayed by ELISA, the Griess method, real-time quantitative PCR, and Western blot and immunocytochemistry method, respectively. The ethanol extract of D. indica not only reduced production of pro-inflammatory cytokines and mediators and blocked NF-kappaB activation, but also slightly promoted release of the anti-inflammatory mediator HO-1 and suppressed IL-10 secretion. In conclusion, the anti-inflammatory effects of the extract of D. indica are attributed to the suppression of pro-inflammatory cytokines and mediators by blocking NF-kappaB activation. The extract of D. indica can also slightly promote HO-1 production to reduce inflammation.
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Anti-Inflamatórios/farmacologia , Citocinas/metabolismo , Medicamentos de Ervas Chinesas/farmacologia , Heme Oxigenase-1/metabolismo , Mediadores da Inflamação/metabolismo , Macrófagos/imunologia , NF-kappa B/metabolismo , Animais , Linhagem Celular , Citocinas/imunologia , Inflamação/imunologia , Inflamação/metabolismo , Macrófagos/citologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , CamundongosRESUMO
Corilagin (beta-1-O-galloyl-3,6-(R)-hexahydroxydiphenoyl-D-glucose) is a novel member of the tannin family which has been discovered from many medicinal plants and has been confirmed in many pharmacological activities. However, the purified Corilagin that was used in experiment is rare, and the anti-inflammatory mechanism of Corilagin has not been investigated clearly. This study is to explore the inner anti-inflammatory mechanism of Corilagin. Inflammatory cellular model was established by lipopolysaccharide (LPS) interfering on RAW264.7 cell line. Levels of TNF-alpha, IL-1beta, IL-6, NO and IL-10 in supernatant, mRNA expression of TNF-alpha, COX-2, iNOS and HO-1, protein expression of COX-2 and HO-1, translocation of NF-kappaB were assayed by ELISA or Griess method, real-time quantitative PCR, western blot and immunocytochemistry method, respectively. As a result, Corilagin could significantly reduce production of pro-inflammatory cytokines and mediators TNF-alpha, IL-1beta, IL-6, NO (iNOS) and COX-2 on both protein and gene level by blocking NF-kappaB nuclear translocation. Meanwhile Corilagin could notably promote release of anti-inflammatory factor HO-1 on both protein and gene level, but suppress the release of IL-10. In conclusion, the anti-inflammatory effects of Corilagin are attributed to the suppression of pro-inflammatory cytokines and mediators by blocking NF-kappaB activation. Corilagin also can promote HO-1 production to induce regression of inflammation but can inhibit IL-10 production like Dexamethasone. Corilagin possesses a potential anti-inflammatory effect by not only abating inflammatory impairment but also promoting regression of inflammation and has a good prospect to be used in many inflammation-related diseases.
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Anti-Inflamatórios/farmacologia , Glucosídeos/farmacologia , Transporte Ativo do Núcleo Celular , Animais , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Citocinas/biossíntese , Heme Oxigenase-1/genética , Taninos Hidrolisáveis , Lipopolissacarídeos/farmacologia , Camundongos , NF-kappa B/metabolismo , Óxido Nítrico/biossíntese , RNA Mensageiro/análiseRESUMO
Kummerowia striata (Thunb.) Schindl has long been used as a fork herb in inflammation-related therapy. This study was undertaken to determine the anti-inflammatory effect of the plant. High performance liquid chromatography (HPLC) was used for evaluating the extract. While dexamethasone (DM) was used as a positive control, the effects of ethanol extract on the production of IL-1beta, IL-6, NO, COX-2 and TNF-alpha, the expression of iNOS mRNA, TNF-alpha mRNA, COX-2 mRNA, protein production of COX-2 and HO-1, NF-kappaB and I-kappaB of LPS-stimulated RAW 264.7 cells were studied by sandwich ELISA, real-time PCR, Western blot analysis and immunocytochemistry assay respectively. The results showed that K. striata (Thunb.) Schindl had a good anti-inflammatory effect on LPS-stimulated RAW264.7 cell. On one hand, it could significantly inhibit the production of IL-1beta, IL-6, NO, TNF-alpha, COX-2 in LPS-stimulated cell than that of single LPS stimulated cell (p < 0.01 or p < 0.05). On the other hand, it could increase the production of IL-10 and HO-1 than that of single LPS intervention cell (p < 0.01 or p < 0.05). Furthermore, the extract also could inhibit the production of NF-kappaB and I-kappaB compared to single LPS stimulated cell. In a word, it suggested that the anti-inflammatory actions of K. striata (Thunb.) Schindl ethanol extract might be due to the down-regulation of IL-1beta, IL-6, NO, TNF-alpha and COX-2 via the suppression of NF-kappaB activation and conversation of I-kappaB production, and another pathway was up regulating the production of IL-10 and HO-1.
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Anti-Inflamatórios/farmacologia , Fabaceae , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/isolamento & purificação , Western Blotting , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Relação Dose-Resposta a Droga , Ensaio de Imunoadsorção Enzimática , Etanol/química , Fabaceae/química , Heme Oxigenase-1/genética , Heme Oxigenase-1/metabolismo , Proteínas I-kappa B/metabolismo , Imuno-Histoquímica , Interleucinas/metabolismo , Macrófagos/enzimologia , Macrófagos/metabolismo , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Camundongos , NF-kappa B/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Componentes Aéreos da Planta , Extratos Vegetais/farmacologia , Reação em Cadeia da Polimerase , Solventes/química , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismoRESUMO
This study is to explore the inner anti-inflammatory mechanism of the ethanol extract of Rungia pectinata (Linn.) Nees. As a result, the ethanol extract of Rungia pectinata (Linn.) Nees could not only strongly reduce production of pro-inflammatory cytokines and mediators via blocking NF-kappaB activation but slightly promote release of anti-inflammatory mediator HO-1 and suppress IL-10 secretion. In conclusion, compared to Dexamethasone, Rungia pectinata (Linn.) Nees has not only similar effects on antagonizing pro-inflammatory mediators and cytokines but also mild effects on promoting production of anti-inflammatory mediators.
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Acanthaceae , Anti-Inflamatórios/farmacologia , Citocinas/antagonistas & inibidores , NF-kappa B/antagonistas & inibidores , Animais , Anti-Inflamatórios/química , Linhagem Celular , Ciclo-Oxigenase 2/metabolismo , Citocinas/genética , Citocinas/metabolismo , Etanol/química , Heme Oxigenase-1/genética , Heme Oxigenase-1/metabolismo , Inflamação/imunologia , Interleucina-10/antagonistas & inibidores , Camundongos , Óxido Nítrico/antagonistas & inibidores , Extratos Vegetais/química , Extratos Vegetais/farmacologiaRESUMO
BACKGROUND: Melilotus suaveolens Ledeb (M. suaveolens Ledeb) has long been used as a folk medicine in inflammation-related therapy. This study was undertaken to determine the anti-inflammatory effect of the plant. METHODS: Petroleum ether fraction, ethyl acetate fraction, n-butanol fraction, aqueous fraction were obtained from ethanol extract of M. suaveolens Ledeb and evaluated by high performance liquid chromatography (HPLC). While dexamethasone (DM) was used as a positive control, the effects of different solution fractions of ethanol extract on tumor necrosis factor alpha (TNF-alpha) mRNA, cyclooxygenase 2 (COX-2) mRNA, COX-2 and nuclear factor kappaB (NF-kappaB) of LPS-stimulated RAW 264.7 cells were studied by real-time PCR, Western blot analysis and immunocytochemical assay, respectively. RESULTS: Coumarin was one of the main ingredients in different solution fractions of ethanol extract except the aqueous fraction with no inflammatory effect. The petroleum ether fraction, ethyl acetate fraction and n-butanol fraction of ethanol extract could inhibit the production of TNF-alpha mRNA, COX-2 mRNA and NF-kappaB to some extent. CONCLUSIONS: Different solution fractions of ethanol extract from M. suaveolens Ledeb had similar anti-inflammatory effect as did dexamethasone except the aqueous fraction. Coumarin was likely to be essential to the anti-inflammatory effect, and other ingredients might attribute to their different anti-inflammatory effects from the HPLC fingerprint.
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Anti-Inflamatórios não Esteroides/farmacologia , Melilotus , Extratos Vegetais/farmacologia , Animais , Linhagem Celular , Ciclo-Oxigenase 2/genética , Lipopolissacarídeos/farmacologia , Melilotus/química , Camundongos , NF-kappa B/antagonistas & inibidores , Extratos Vegetais/análise , RNA Mensageiro/análise , Fator de Necrose Tumoral alfa/genéticaRESUMO
UNLABELLED: Melilotus suaveolens Ledeb is a species of traditional medical plant for treating inflammation-related disease. This study is to explore the inner anti-inflammatory mechanism on petroleum ether extract from Melilotus suaveolens Ledeb. MATERIALS AND METHODS: Inflammatory cellular model was founded by intervention of lipopolysaccharide (LPS) on RAW264.7 cell line. Secretion of TNF-alpha, IL-1beta, IL-6, NO and IL-10 in supernatant, mRNA expression of TNF-alpha, COX-2, iNOS and HO-1, protein expression of COX-2 and HO-1, activation of NF-kappaB and ingredients in the extract were assayed. RESULTS: The extract could not only reduce production of pro-inflammatory mediators by blocking NF-kappaB activation but promote release of anti-inflammatory mediator HO-1 significantly. The only active ingredient in the extract was coumarin and the concentration of coumarin in each 1 g extract was 0.27822 mg. CONCLUSION: Compared to Dexamethasone, the extract not only has similar effects on antagonizing pro-inflammatory mediators and cytokines but has effects on promoting production of anti-inflammatory mediators.