RESUMO
AIM To establish an HPLC method for the simultaneous content determination of five constituents in Simotang Oral Liquid (Aucklandiae Radix,Aurantii Fructus,Arecae Semen,etc.).METHODS The analysis of 50% methanol extract of this drug was performed on a 35 ℃ thermostatic Agilent Zorbax C18 column (4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1% formic acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 283 nm.RESULTS Synephrine,norisoboldine,naringin,hesperidin and neohesperidin showed good linear relationships within the ranges of 5.8-185.6 μg/mL (r =0.999 9),0.829-26.52 μg/mL (r =1.000 0),9.775-312.8 μg/mL (r =1.000 0),0.594-19 μg/mL (r =0.999 5) and 5.2-166.4 μg/mL (r =1.000 0),whose average recoveries were 98.93%,98.95%,102.57%,99.67% and 103.43% with the RSDs of 1.85%,1.27%,0.52%,0.89% and 0.43%,respectively.CONCLUSION This simple,accurate and reproducible method can be used for the rapid quality control of Simotang Oral Liquid.
RESUMO
Although the etiology of inflammatory bowel disease is still uncertain, increasing evidence indicates that the excessive activation of NLRP3 inflammasome plays a major role. Norisoboldine (NOR), an alkaloid isolated from Radix Linderae, has previously been demonstrated to inhibit inflammation and IL-1β production. The present study was to examine the effect of NOR on colitis and the underlying mechanism related to NLRP3 inflammasome activation. Our results showed that NOR alleviated colitis symptom in mice induced by 2, 4, 6-trinitrobenzene sulfonic acid (TNBS). Moreover, it significantly reduced expressions of cleaved IL-1β, NLRP3 and cleaved Caspase-1 but not ASC in colons of mice. In THP-1 cells, NOR suppressed the expressions of NLRP3, cleaved Caspase-1 and cleaved IL-1β but not ASC induced by lipopolysaccharide (LPS) and adenosine triphosphate (ATP). Furthermore, NOR could activate aryl hydrocarbon receptor (AhR) in THP-1 cells, inducing CYP1A1 mRNA expression, and promoting dissociation of AhR/HSP90 complexes, association of AhR and ARNT, AhR nuclear translocation, XRE reporter activity and binding activity of AhR/ARNT/XRE. Both siAhR and α-naphthoflavone (α-NF) markedly diminished the inhibition of NOR on NLRP3 inflammasome activation. In addition, NOR elevated Nrf2 level and reduced ROS level in LPS- and ATP-stimulated THP-1 cells, which was reversed by either siAhR or α-NF treatment. Finally, correlations between activation of AhR and attenuation of colitis, inhibition of NLRP3 inflammasome activation and up-regulation of Nrf2 level in colons were validated in mice with TNBS-induced colitis. Taken together, NOR ameliorated TNBS-induced colitis in mice through inhibiting NLRP3 inflammasome activation via regulating AhR/Nrf2/ROS signaling pathway.
Assuntos
Animais , Humanos , Masculino , Camundongos , Alcaloides , Colite , Tratamento Farmacológico , Genética , Alergia e Imunologia , Medicamentos de Ervas Chinesas , Inflamassomos , Alergia e Imunologia , Interleucina-1beta , Genética , Alergia e Imunologia , Lindera , Química , Camundongos Endogâmicos BALB C , NF-kappa B , Genética , Alergia e Imunologia , Receptores de Hidrocarboneto Arílico , Genética , Metabolismo , Ácido TrinitrobenzenossulfônicoRESUMO
Although the etiology of inflammatory bowel disease is still uncertain, increasing evidence indicates that the excessive activation of NLRP3 inflammasome plays a major role. Norisoboldine (NOR), an alkaloid isolated from Radix Linderae, has previously been demonstrated to inhibit inflammation and IL-1β production. The present study was to examine the effect of NOR on colitis and the underlying mechanism related to NLRP3 inflammasome activation. Our results showed that NOR alleviated colitis symptom in mice induced by 2, 4, 6-trinitrobenzene sulfonic acid (TNBS). Moreover, it significantly reduced expressions of cleaved IL-1β, NLRP3 and cleaved Caspase-1 but not ASC in colons of mice. In THP-1 cells, NOR suppressed the expressions of NLRP3, cleaved Caspase-1 and cleaved IL-1β but not ASC induced by lipopolysaccharide (LPS) and adenosine triphosphate (ATP). Furthermore, NOR could activate aryl hydrocarbon receptor (AhR) in THP-1 cells, inducing CYP1A1 mRNA expression, and promoting dissociation of AhR/HSP90 complexes, association of AhR and ARNT, AhR nuclear translocation, XRE reporter activity and binding activity of AhR/ARNT/XRE. Both siAhR and α-naphthoflavone (α-NF) markedly diminished the inhibition of NOR on NLRP3 inflammasome activation. In addition, NOR elevated Nrf2 level and reduced ROS level in LPS- and ATP-stimulated THP-1 cells, which was reversed by either siAhR or α-NF treatment. Finally, correlations between activation of AhR and attenuation of colitis, inhibition of NLRP3 inflammasome activation and up-regulation of Nrf2 level in colons were validated in mice with TNBS-induced colitis. Taken together, NOR ameliorated TNBS-induced colitis in mice through inhibiting NLRP3 inflammasome activation via regulating AhR/Nrf2/ROS signaling pathway.
Assuntos
Animais , Humanos , Masculino , Camundongos , Alcaloides , Colite , Tratamento Farmacológico , Genética , Alergia e Imunologia , Medicamentos de Ervas Chinesas , Inflamassomos , Alergia e Imunologia , Interleucina-1beta , Genética , Alergia e Imunologia , Lindera , Química , Camundongos Endogâmicos BALB C , NF-kappa B , Genética , Alergia e Imunologia , Receptores de Hidrocarboneto Arílico , Genética , Metabolismo , Ácido TrinitrobenzenossulfônicoRESUMO
Objective:To establish an HPLC method for the determination of norisoboldine in Suoquan capsules. Methods: The chromatographic procedure with acetonitrile-water containing 0. 5% folic acid and 0. 1% triethylamine (10∶90) as the mobile phase was carried out on an Ecosil C18 HPLC(150 mm ×4.6 mm,5 μm). The flow rate was at 1.0 ml·min-1, the detection wavelength was 280nm and the column temperature was 30 ℃. Results: The linear range of norisoboldine was within 38. 56-578. 40 ng ( r =0. 999 2), the average recovery of norisoboldine was 99. 9%(RSD=1. 7%, n=6). Conclusion:The method is highly sensitive, re-liable and accurate, and can be applied in the determination of Suoquan capsules.