ABSTRACT
Objective To determine whether store-operated Ca2+ entry (SOCE) is involved in chronic hypoxia-induced alteration of intracellular Ca2 + concentration ([Ca2+] i) and proliferation in pulmonary arterial smooth muscle cells (PASMC).Methods Rat PASMCs were cultured and treated in normoxia (21％O2) or hypoxia (4％ O2) condition.The proliferation of PASMC was detected by cell counting kit-8 (CCK-8) assay.[Ca2 +] i,SOCE and the effects of store-operated Ca2 + channel (SOCC) inhibitors,SKF96365 and NiCl2,on SOCE in hypoxic PASMCs were tested by InCyte [Ca2 +] i measurement system.Results Hypoxia for 24-60 h augmented PASMC proliferation (1.12 ± 0.09 vs 0.71 ± 0.05,P ＜ 0.05) and [Ca2 +] i [(214.8 ± 20.4) nmol/L vs (115.2 ± 13.2) nmol/L,P ＜ 0.05] in a time-dependent manner with the maximum effect at 60 h.Perfusion of Ca2+-free Krebs solution containing nifedipine (5 μ mol/L),cyclopiazonic acid (CPA,10 μmol/L) in PASMCs caused a small transient increase of [Ca2+]i with peak [Ca2+]i (113.3 ± 49.3) nmol/L.Chronic hypoxia (4％ O2,60 h) enhanced [Ca2+]i level with peak value of (193.2 ± 22.7) nmol/L (P ＜ 0.05) in PASMC.After restoration of extracellular Ca2+,CPA caused marked increase of [Ca2+]i with peak value of (328.0 ± 56.7) nmol/L.Chronic hypoxia strengthened CPA-induced increase of [Ca2 +] i with peak value of (526.0 ± 33.7) nmol/L (P ＜ 0.05) in PASMCs.Either SKF96365 50 μmol/L or NiCl2 500 μmol/L distinctly attenuated CPA-induced enhancement of [Ca2 +] i,the peak value of which dropped from (526.0 ± 33.7) nmol/L to (170.4 ± 26.4) nmol/L (P＜0.05) or (177.4±45.9) nmol/L (P＜0.05) respectively.Conclusion Chronic hypoxia boosts the release of Ca2+ from sarcoplasmic reticulum and promotes the activity of SOCC and SOCE,leading to [Ca2 +] i elevation and proliferation of rat PASMCs.
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Objective To establish an analytical method for determination of ferulic acid inDanggui HujiaoRadix Angelicae Sinensis and Fructus Piperis]) Decoction in mouse plasma.Methods HPLC method was used.The conditions of chromatography: Kromasil C 18250 mm?4.6 mm, 7 ?m) was used with a mobile phase of CH3OH-H2O-CH3COOH (36.4∶63∶0.6).Flow rate was 1.0 mL/min.The detecting wavelength was 322 nm.External standard method was quantitative analysis method.Results The ferulic acid could be totally separated from other ingredients in plasma.The linear range was 1.88—188.00 ng/?L (r=0.999), the lowest detectability was 0.47 ng/?L, and the average recovery was 94.85%.Conclusion This method provides an accurate and sensitive way in detecting blood concentration of ferulic acid and studying in pharmacokinetics.
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Objective An HPLC method was established for determination of paeoniflorin in plasma after ig compound decoction of Radix Paeoniae Rubra with Fructus Piperis to mice. Methods The conditions of chromatography: Kromasil C 18 column (250 mm ? 4.6 mm, 7 ?m) was used with a mobile phase of CH 3OH-H 2O ( 38∶ 62); flow rate was 0.5 mL/min; the detecting wavelength was 230 nm; external standard method was quantitative analysis method. Results Paeoniflorin was fully separated from the other components in plasma. The linear range was 5.0—250.0 ng/?L (r= 0.999 9 ), the lowest detectability was 1.49 ng/?L, the average recovery was higher than 90%. Conclusion This method specially provides an accurate and sensitive way in detecting the in vivo blood concentration of paeoniflorin in plasma.
ABSTRACT
Objective To establish a RP-HPLC method for the determination of the blood concentration of paeoniflorin which was produced by seven kinds of warming-inside drugs (WID) being used with the promoting-blood drugs (PBD) individually and to explore the mechanism of PBD and WID compound prescription. Methods The blood concentration of peaoniflorin in mouse plasma was determined by HPLC after ig seven kinds of WID being compatible with Radix Paeoniae Rubra (RPR) to mice separately. Results Fructus Piperis (FP), Cortex Cinnamoni (CC), Fructus Evodiae (FE), Fructus Foeniculi (FF), and Pericarpium Zanthoxyli (PZ) compound prescription with RPR can increase the blood concentration of paeoniflorin in different degrees (P