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.

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.

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.