ABSTRACT
Objective To observe the effect of sinomenine (SIN) on the expression of cyclooxygenase (COX2),alpha-7 nicotinic acetylcholine receptor(α7nAChR) and adenosine receptor(A2A) in A549 cells,and to explore the relative mechanism for cell proliferation.Methods The effect of SIN and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) on the proliferation of A549 cells was determined by methyl thiazolyl tetrazolium (MTT) assay.The effect of SIN and NNK on the migration of A549 cells was detected by cell wound scratch assay.The effect of SIN and NNK on COX2 expression in A549 cells was determined by Western blotting method.The effect of SIN and NNK on the expression of α7nAChR and A2A mRNA and protein was evaluated by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting method.Results NNK increased the proliferation and migration of A549 cells,while SIN inhibited the proliferation and migration of A549 cells.COX2 expression level was increased in NNK group but was decreased in SIN group.The expression levels of α7nAChR and A2A were up-regulated in NNK group but were down-regulated in SIN group.Conclusion SIN plays a role in inhibiting the proliferation and migration of A549 cells by suppressing COX2 expression.SIN has an inhibitory effect on the expression of α7nAChR and A2A.
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Objective To investigate the effect of sinomenine on the purinergic receptors A2A and P2X7 in endotoxemia mouse model and RAW264.7 macrophage model stimulated by lipopolysaccharide(LPS). Methods BALB/c mice were randomly divided into blank control group, model group and sinomenine group. Thirty minutes after the rats of sinomenine group were pretreated with intraperitoneal injection of sinomenine (40, 80, 160 mg/kg), the mice were given intraperitoneal injection of 15 mg/kg LPS to induce endotoxemia model. The serum levels of tumor necrosis factor-alpha(TNF-α) and interleukin-6(IL-6) were measured by enzyme-linked immunosorbent assay (ELISA). The expression levels of purinergic receptor A2A and P2X7 in the liver and spleen were detected by reverse transcription-polymerase chain reaction(RT-PCR). RAW264.7 macrophages were divided into blank control group, LPS group and sinomenine group. Sinomenine group was firstly treated with sinomenine(300μmol/L) for 2 h, and then LPS group and sinomenine group were treated with LPS (100 ng/mL) for another 8 hours. TNF-α in the cell supernatant was measured by ELISA, and the expression levels of A2A and P2X7 in RAW264.7 cells were detected by RT-PCR. Results Stimulation with LPS could induce the increase of the mouse serum levels of TNF-α and IL-6 as well as the expression of A2A and P2X7 in mouse liver and spleen, and sinomenine had a counteraction on the above indexes(P<0.05) . In-vitro stimulation with LPS could induce the increase of the content of TNF-α and the expression of A2A and P2X7 in RAW264.7 cells , and sinomenine decreased TNF-α content and P2X7 expression (P<0.05) , but had an effect on enhancing A2A expression. Conclusion Sinomenine suppresses the expression of purinergic receptor P2X7 in mouse spleen and liver as well as in RAW264.7 macrophages, but its effect on the expression of A2A in various tissues and cells varies, whose related mechanism is needed further study.
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Objective To observe the effect of sinomenine(SINO) on immune function of adjuvant arthritis(AA) rats.Methods SD rats were randomized into normal group,model group,SINO groups(treated with gastric gavage of SINO at the doses of 60,120 and 240 mg/kg respectively),and glucosides of Tripterygium Wilfordii(30 mg/kg) group.Except the normal group,the rats in other groups received subcutaneous injection of the complete Freund's adjuvant 0.1mL into the left hind foot to induce AA.The medication began from 12 days after the modeling and lasted 12 days.The pedal swelling and joint function scores were observed in different time.Radioimmunoassay was used to detect the contents of interleukin-1?(IL-1?) and tumor necrosis factor ?(TNF-?) in synovial cells.Expression of IL-1? and TNF-? mRNA was examined by reverse transcription-polymerase chain reaction(RT-PCR) assay.Results SINO at different concentrations decreased the pedal swelling and arthritis scores to various degrees,inhibited the production of IL-1? and TNF-? in the synovial cells,reduced the expression of IL-1? and TNF-? mRNA,and recovered the normal histological features of synovial cells in AA rats.Conclusion The therapeutic mechanism of SINO for rheumatoid arthritis may be related with the inhibition of secretion of inflammatory mediators in synovial cells,and with the recovery of histological features of synovial cells.
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[Objective] To explore the anti - inflammatory mechanism of sinomenine, a monomer alkaloid from Caulis Sinomenii. [ Methods] In - vitro enzyme was applied to observe the effect of sinomenine on cyclooxygenase (COX) 1 and COX 2. [Results] Sinomenine at the concentration of 1 - 250 ?mol/L could inhibit the formation of PGE2 induced by COX 2 in a good dose - effect manner. [Conclusion] Sinomenine selectively inhibiting the activity of COX 2 may be the anti - inflammatory and anagesic mechanism of sinomenine with less gastrointestinal adverse effect.
ABSTRACT
【Objective】The effect of sinomenine on human cyclo-oxygenase(COX) and interleukin-1?(IL-1?) was observed to explore the anti-inflammatory mechanism of sinomenine.【Methods】Peripheral blood monocytes(PBMC) were isolated and incubated with sinomenine.After incubation,total RNA from PBMC was extracted and the effect of sinomenine on COX and IL-1? expression with or without the stimulation of lipopolysaccharide(LPS) was observed by the method of semi-quantitative reverse transcription polymerase chain reaction(RT-PCR).【Results】Sinomenine had no effect on COX-1 gene expression,had a slight inhibitory effect on COX-2 gene expression and had a strong inhibitory effect on IL-1? gene expression.【Conclusion】 The anti-inflammatory mechanism may be related to the inhibition of inflammatory cytokines.