ABSTRACT
OBJECTIVE： To investigate the vasodilatory effect mechanism of psoralen and bakuchiol. METHODS： The rat thoracic aorta was isolated to prepare vascular rings and de-endothelium vascular rings. Using contraction rate as index， the intact endothelium or de-endothelium vascular rings were pre-incubated with N-nitro-L-arginine methyl ester （L-NAME， 100 μmol/L）； vasodilatory effect of low-dose， medium-dose and high-dose of psoralen or bakuchiol（0.1，1，10 μmol/L）on aortic vessels pre- contracted with norepinephrine （NE， 1 μmol/L） or potassium chloride （KCl， 60 mmol/L） were investigated. The de-endothelium vascular rings were pre-incubated with calcium dependent potassium channel inhibitors tetraethylammonium chloride （TEA， 0.1 mmol/L） and inward rectifying potassium channel inhibitor barium chloride （BaCl2，0.1 mmol/L）； vasodilatory effect of low-dose， medium-dose and high-dose of bakuchiol （0.1， 1， 10 μmol/L） on de-endothelium vascular vessels pre-contracted with NE （1 μmol/L） were investigated. The microvascular endothelial cells were isolated by collagenase-neutral protease digestion； the effects of low-dose， medium-dose and high-dose of psoralen or bakuchiol （0.1， 1， 10 μmol/L） on the expression of eNOS protein were studied by ELISA. RESULTS： Psoralen and bakuchiol could significantly reduce the contraction rate of endothelium-intact aortic rings pre-contracted with NE（P＜0.01）； medium-dose and high-dose of psoralen and bakuchiol could significantly reduce the contraction rate of endothelium-intact aortic rings pre-contracted with KCl（P＜0.05 or P＜0.01）； while the contraction rate could be increased by de-endothelium and NOS inhibition significantly （P＜0.05 or P＜0.01）. The medium-dose and high-dose of bakuchiol could significantly reduce the contraction rate of de-endothelium vascular vessels pre-contracted with NE （P＜0.05 or P＜0.01）. The contraction rate could be increased by inhibiting inward rectifier potassium channels in vascular smooth muscle （P＜0.01）. Different dosages of psoralen and bakuchiol could significantly increase the expression levels of eNOS protein in rat cardiac microvascular endothelial cells（P＜0.01）. CONCLUSIONS： Psoralen and bakuchiol may play a role in vasodilation via endothelium-dependent NO pathway and by promoting eNOS protein expression in endothelial cells； bakuchiol may play a role in vasodilation via non-endothelium dependent pathway as opening inward rectifying potassium channel.
ABSTRACT
OBJECTIVE: To study the effects of ultrafine grinding technology on the dissolution of triterpenoids from Ganoderma lucidum.METHODS: UV spectrophotometry was used to determine total extration rates of triterpenoids from G. lucidum. HPLC method was used to determine the contents of 9 kinds of triterpenoids (ganoderic acid A, ganoderic acid B, ganoderic acid C, ganoderic acid C1, ganoderic acid C2, ganoderic acid D, ganoderic acid E, ganoderic acid G, ganoderic acid H) from G. lucidum. The total extration rates and the contents of 9 kinds of triterpenoids were compared between 30, 50, 80 mesh common G. lucidum powder and 300 mesh G. lucidum ultramicro powder. RESULTS: The total extraction rates of triterpenoids from 30, 50, 80, 300 mesh G. lucidum powder were (0. 74 ± 0. 08)％,(0. 75 ± 0. 06)％,(0. 78 ± 0. 06)％,(1. 09 ± 0. 10)％ (RSD< 2％, n=3), respectively. With the increase of the mesh number of G. lucidum powder, the contents of triterpenoids were increased gradually, and were the highest in 300 mesh, which was significant higher than common G. lucidum powder (P<0. 05). CONCLUSIONS: After ultrafine grinding, the dissolution of triterpenoids from G. lucidum is increased.
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OBJECTIVE:To optimize the best crushing particle size of Agkistrodon decocting powder,and study its effect on apoptosis of human rheumatoid arthritis fibroblast-like synoviocytes. METHODS:Pre-column derivatization RP-HPLC was adopted. Using the total decoction amounts of 4 main amino acids (aspartic acid,glutamic acid,L-hydroxyproline,glycine) that decocted once as index,the Agkistrodon decoction pieces and those through No.1-8 seive were screened,and the best crushing particle size of Agkistrodon decocting powder was optimized. The human rheumatoid arthritis fibroblast-like synoviocytes were divided into nega-tive control group,positive control group (1 μmol/L methotrexate) and the best crushing particle size of Agkistrodon decocting powder group(2.0 mg/mL),flow cytometry was used to determine the cell apoptosis after cultured for 48 h. RESULTS:The best crushing particle size was through No.6 seive,when the total amount of 4 main amino acids was(61.27±0.02)mg/g(n=3). Com-pared with negative control group,the apoptosis rate in the best crushing particle size of Agkistrodon decocting powder group was significantly increased(P<0.05),which was slightly higher than positive control group. CONCLUSIONS:The best crushing parti-cle size is through No.6 seive;and Agkistrodon decocting powder shows effect on reducing the apoptosis of human rheumatoid ar-thritis fibroblast-like synoviocytes.
ABSTRACT
OBJECTIVE:To optimize the best crushing particle size of Agkistrodon decocting powder,and study its effect on apoptosis of human rheumatoid arthritis fibroblast-like synoviocytes. METHODS:Pre-column derivatization RP-HPLC was adopted. Using the total decoction amounts of 4 main amino acids (aspartic acid,glutamic acid,L-hydroxyproline,glycine) that decocted once as index,the Agkistrodon decoction pieces and those through No.1-8 seive were screened,and the best crushing particle size of Agkistrodon decocting powder was optimized. The human rheumatoid arthritis fibroblast-like synoviocytes were divided into nega-tive control group,positive control group (1 μmol/L methotrexate) and the best crushing particle size of Agkistrodon decocting powder group(2.0 mg/mL),flow cytometry was used to determine the cell apoptosis after cultured for 48 h. RESULTS:The best crushing particle size was through No.6 seive,when the total amount of 4 main amino acids was(61.27±0.02)mg/g(n=3). Com-pared with negative control group,the apoptosis rate in the best crushing particle size of Agkistrodon decocting powder group was significantly increased(P<0.05),which was slightly higher than positive control group. CONCLUSIONS:The best crushing parti-cle size is through No.6 seive;and Agkistrodon decocting powder shows effect on reducing the apoptosis of human rheumatoid ar-thritis fibroblast-like synoviocytes.
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OBJECTIVE:To optimize the decoction process of agkistrodon. METHODS:RP-HPLC with pre-column derivatiza-tion was adopted. With the contents of 4 main amino acids in agkistrodon as index,the decoction process(decocting times,heating time,water quantity and medicinal material crushing granularity) was optimized by orthogonal tests and verified. RESULTS:The optimal technology of the decoction process of agkistrodon was as 3 times of decoction,60 min of heating time,50 ml of water consumption for 0.90 g medicinal material and No.6 sieve. The results of verification test showed the total extraction of 4 kinds of amino acids was 72.68 mg/g(RSD=3.77%,n=3). CONCLUSIONS:The decoction technology can be used for the decoction pro-cess of agkistrodon,and it is stable and feasible.
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To investigate the vasodilator effect and the endothelium-dependent mechanism of oroxylin A in thoracic aorta isolated from rats.