ABSTRACT
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.
ABSTRACT
Objective To study the relationship between the inflammatory cytokine levels and the expression of hepatic alpha-7 nicotinic acetylcholine receptor (α7 nAChR) in hyperlipidemia rats and to investigate the intervention effect of Yirui Capsules ( mainly composed of Radix Salviae Miltiorrhizae, Fructus Crataegi, Rhizoma Alismatis, etc). Methods Fifty male SD rats were randomly divided into blank control group ( fed with conventional diet, N=10) and high-fat diet group (fed with high-fat diet, N=40). After feeding for 14 d, the forty rats in high-fat diet group were further randomly divided into model control group, and low-, middle-, and high-dose Yirui Capsules groups (140, 280, 560 mg/kg, respectively), 10 rats in each group and the treatment lasting 30 days. And then, the serum levels of lipids were detected, C-reactive protein (CRP) and tumor necrosis factor (TNF) -α in the serum were detected by enzyme-linked immunosorbent assay ( ELISA) , and the expression of hepatic α7 nAChR was analyzed by immunohistochemical staining and reverse transcription-polymerase chain reaction (RT-PCR) . Results Yirui Capsules in different dosages could improve serum lipid levels in hyperlipidemia rats to various degrees. The contents of serum inflammatory cytokines CRP and TNF-α and the mRNA expression of hepatic α7nAChR were increased in the model control group as compared with those in the blank control group ( P<0.01). The contents of inflammatory cytokines CRP, TNF-αand the mRNA expression of hepatic α7nAChR were decreased in the three Yirui Capsules groups as compared with those in the model control group ( P<0.01). The results of immunohistochemical staining for α7nAChR expression were consistent with RT-PCR results. Conclusion The expression of hepatic α7nAChR is increased in hyperlipidemia rats. Yirui capsules are effective on decreasing inflammatory cytokine levels, improving lipid metabolism, and down-regulating α7nAChR expression.
ABSTRACT
Methods The maximum tolerated dose( MTD)of water and ethanol extracts of Radix millettiae speciosae were measured for the safety evaluation. Results Maximal tolerated dose( MTD)of water extract was higher than 1 000 g·kg-1 ,which equaled to 110 times the dose for human adults(60 kg). MTD of ethanol extract was higher than 1 700 g·kg-1 ,which equaled to 186 times the dose for human adults(60 kg). Conclusion Radix millettiae speciosae does not have obvious toxicity,thus its routine clinical dose is safe and feasible.