右归饮及其入血成分对大鼠成骨细胞的促增殖作用

Proliferative Effect of Youguiyin and its Main Constituents Absorbed in the Blood on Rats Osteoblasts

目的明确右归饮的主要入血成分;阐明右归饮及其主要入血成分对大鼠成骨细胞的促增殖作用。方法应用超高效液相色谱-质谱联用(UPLC-MS)技术建立右归饮、各单味药及Sprague-Dawley大鼠灌服右归饮后含药血清的分析方法;体外分离培养成骨细胞,噻唑蓝(MTT)法评价右归饮、含药血清及其主要入血成分对成骨细胞增殖率的影响。结果大鼠灌服右归饮后在含药血清中检测到10种入血成分,分别为君药附子中的乌头碱、次乌头碱、新乌头碱、苯甲酰乌头原碱、苯甲酰次乌头原碱、苯甲酰新乌头原碱,君药肉桂中的肉桂酸,臣药山茱萸中的莫诺苷、马钱苷和佐药杜仲中的松脂醇二葡萄糖苷。右归饮药液及两味君药附子、肉桂中7种入血成分浓度分别为1μg·mL^(-1)、10 ng·mL^(-1)、10 ng·mL^(-1)、1 ng·mL^(-1)、100μg·mL^(-1)、100 pg·mL^(-1)、1μg·mL^(-1)和1 ng·mL^(-1)时,对成骨细胞有一定的促增殖作用且起效较为迅速,右归饮含药血清的起效时间缓慢。结论右归饮可原型入血的10种化学成分得以揭示,且在一定浓度范围内,右归饮药液、右归饮含药血清及右归饮主要入血成分对成骨细胞具有促增殖作用。

Objective To identify the main blood absorbed constituents from Youguiyin and elucidate the proliferative effect of Youguiyin and its main blood absorbed constituents on osteoblasts in rats.Methods Ultra-performance liquid chromatography-mass spectrometry(UPLC-MS)technique was used to establish the analysis method for Youguiyin,individual herbal medicines and the medicated serum containing Youguiyin after oral administration in Sprague-Dawley rats.Osteoblasts of rats were isolated and cultured in vitro.The effect of Youguiyin,medicated serum,and its main blood absorbed components on osteoblast proliferation rate was evaluated by thiazole blue(MTT)assay.Results Ten components were detected in the blood serum of rats after oral administration of Youguiyin,namely aconitine,hypaconitine,mesaconitine,benzoylaconitine,benzoylhypaconitine,benzoylmesaconine from Aconiti Lateralis Radix Praeparata,cinnamic acid from Cinnamomi Cortex,morroniside,loganin from Corni Fructus,and pinoresinoldiglucoside from Eucommiae Cortex,respectively.Youguiyin decoction and seven components from Aconiti Lateralis Radix Praeparata and Cinnamomi Cortex showed a certain promoting proliferation effect on osteoblasts when the concentrations were 1μg·mL^(-1),10 ng·mL^(-1),10 ng·mL^(-1),1 ng·mL^(-1),100μg·mL^(-1),100 pg·mL^(-1),1μg·mL^(-1)and 1 ng·mL^(-1),respectively.Besides,the effect onset was relatively rapid.The onset time of medicated serum containing Youguiyin was slow.Conclusions The ten chemical components absorbed in the blood from Youguiyin were identified.It was revealed that Youguiyin,medicated serum and the main components of Youguiyin absorbed in the blood can promote the proliferation of osteoblasts within a certain concentration range.