淡豆豉炮制中γ-氨基丁酸含量测定及其抗抑郁作用研究

Determination of content ofγ-aminobutyric acid and its antidepressant effect at different time points during processing of Sojae Semen Praeparatum

目的检测淡豆豉炮制过程中各样本γ-氨基丁酸(GABA)含量并研究其抗抑郁作用。方法应用柱前在线衍生-高效液相色谱技术测定原料黑大豆(H)、发酵第6天(F6)、再闷6 d(Z6)、再闷15 d(Z15)样本水煎液中GABA含量;将昆明种小鼠随机分成对照组、模型组、盐酸氟西汀(阳性对照,0.01 g/kg)组、GABA(0.0128 g/kg)组、H(1.52 g/kg)组、F6(1.52 g/kg)组、Z6(1.52 g/kg)组和Z15低、中、高剂量(0.76、1.52、3.04 g/kg)组,采用单笼饲养加慢性温和不可预知性应激(CUMS)制备小鼠抑郁模型,各组在造模同时开始ig给药,每日应激前1 h给药,连续28 d。检测各组小鼠体质量、糖水偏好、敞箱、悬尾、强迫游泳等行为学指标的变化。结果H和F6样本水煎液未检出GABA,Z6、Z15样本的GABA质量分数分别为5.559、8.421 mg/g。与对照组比较,模型组小鼠的体质量、糖水偏好、跨格数和直立数均显著减少(P<0.01),悬尾及游泳不动时间显著增加(P<0.05、0.01),表明CUMS抑郁模型制备成功。与模型组比较,盐酸氟西汀组、GABA组、Z6组和Z15高剂量组糖水偏爱度、敞箱实验跨格数和直立数均显著增加(P<0.05、0.01),Z15中剂量组糖水偏爱度和敞箱实验直立数显著增加(P<0.05、0.01);盐酸氟西汀组、GABA组、Z6组及Z15高、中剂量组悬尾实验的不动时间均显著减少(P<0.05、0.01);各组小鼠强迫游泳实验的不动时间均显著缩短(P<0.05、0.01)。结论发现淡豆豉炮制后期出现高含量GABA;淡豆豉显著改善小鼠的快感缺失、行为绝望等抑郁症状,具有良好的抗抑郁作用并与GABA含量可能有关。

Objective To study the content ofγ-aminobutyric acid(GABA)and its antidepressant effect of each sample at different time points during the processing of Sojae Semen Praeparatum(SSP).Methods The GABA content in the decoction of raw black soybean(H),fermentation day 6(F6),"secondary fermentation"for 6 days(Z6)and 15 days(Z15)was determined by pre-column on-line high performance liquid chromatography(HPLC).KM mice were randomly divided into control group,model group,fluoxetine hydrochloride(positive control,0.01 g/kg)group,GABA(0.0128 g/kg)group,H(1.52 g/kg)group,F6(1.52 g/kg)group,Z6 group(1.52 g/kg)and Z15 low,high,medium and doses(0.76,1.52 and 3.04 g/kg).The mouse depression model was prepared by single cage feeding plus chronic mild unpredictable stress,each at the same time in the building began to ig drugs.The drug was given 1 h before daily stress for 28 consecutive days.The body weight,sugar water preference and open box of each group were detected.Changes in behavioral indicators such as hangovers and forced swimming were detected.Results GABA content in each sample decoction:H and F6 were not detected,and the content of Z6 and Z15 were 5.559,8.421 mg/g.Antidepressant effects of each sample:Compared with control group,the weight,syrup preference,cross-grid number and erect number of model group were significantly decreased(P<0.01),and the time of suspension and swimming was significant.An increase(P<0.05 or 0.01)indicates that the CUMS depression model was successfully constructed.Compared with model group,the preference for sugar water,the cross-cell number and the upright number of open box experiment in metformin hydrochloride group,GABA group,Z6 group and Z15 high-dose group were significantly increased(P<0.05 or 0.01),and the preference for sugar water and the upright number of open box experiment in Z15 medium-dose group were significantly increased(P<0.05 or 0.01).The immovable time of tail suspension test in metformin hydrochloride group,GABA group,Z6 group and Z15 high-dose and medium-dose groups was significantly decreased(P<0.05 or 0.01).The immobility time of forced swimming test was significantly shortened in all groups(P<0.05 or 0.01).Conclusion It was found that there was high content of GABA in the later stage of processing of Sojae Semen Praeparatum,Sojae Semen Praeparatum significantly improved the depression symptoms of mice,such as lack of pleasure and behavior despair,which had good antidepressant effect and might be related to the content of GABA.