柴胡加龙骨牡蛎汤对抑郁症肝郁气滞型大鼠肝脏sEH表达的影响

Effect of Chaihu Jia Longgu Muli Decoction(柴胡加龙骨牡蛎汤)on sEH Expression in Liver of Rats with Liver Depression and Qi Stagnation Syndrome of Depression

目的基于肝脏可溶性环氧化物水解酶(soluble epoxide hydrolase,sEH)表达变化探讨柴胡加龙骨牡蛎汤对肝郁大鼠的影响。方法选取50只SPF级SD雄性大鼠,10只作为空白组,40只建立慢性温和不可预知性刺激(CUMS)大鼠模型,成模后随机分为模型组、氟西汀组、柴胡加龙骨牡蛎汤低剂量组、柴胡加龙骨牡蛎汤高剂量组,每组各10只。连续灌胃给药4周,1次/d,分别于造模前、成模后及给药后进行旷场实验、悬尾实验、强迫游泳实验评估大鼠抑郁状态,实时荧光定量聚合酶链式反应(RT-PCR)检测大鼠肝脏组织中sEH mRNA表达,蛋白免疫印迹法(Western blot)定量检测大鼠肝脏组织中sEH蛋白表达。结果与空白组比较,模型组大鼠活动总路程、悬尾挣扎时间均明显缩短(P<0.05),游泳不动时间明显延长(P<0.05),肝sEH蛋白含量及mRNA表达水平均明显升高(P<0.05)。与模型组比较,氟西汀组、柴胡加龙骨牡蛎汤低剂量组(CLMT-L)、柴胡加龙骨牡蛎汤高剂量组(CLMT-H)的活动总路程、悬尾挣扎时间均明显延长(P<0.05),游泳不动时间均明显缩短(P<0.05),肝sEH蛋白含量及mRNA表达水平均明显降低(P<0.05)。干预后,氟西汀组、CLMT-L组、CLMT-H组活动总路程、悬尾挣扎时间均明显长于干预前(P<0.05),游泳不动时间明显短于干预前(P<0.05)。结论抑郁发生后可以增加sEH在肝脏内的表达,而柴胡加龙骨牡蛎汤能显著改善肝郁气滞大鼠的抑郁样行为,并降低肝脏sEH的表达,其作用机制可能与肝sEH含量变化有关。

Objective To investigate the influence of Chaihu Jia Longgu Mulitang(柴胡加龙骨牡蛎汤,CLMD)on liver depression rats based on the change of soluble epoxide hydrolase(sEH)expression. Methods Among 50 SPF male SD rats,10 were randomly selected as normal group and the rest were established chronic unpredictable mild model. The successfully modeled rats were randomly divided into model group,fluoxetine hydrochloride group,CLMD-L group and CLMD-H group with 10 rats in each group. Behavioral changes of rats in each group were evaluated by open field test,tail suspension test,and forced swimming test before,after modeling,and after 4-week treated with CLMD,while sEH mRNA expression in rat liver was detected by Real-time PCR,sEH protein expression in rat liver was quantitatively detected by Western blot. Results Compared with the normal group,the total walking distance and tail hanging struggle time of rats in the model group were significantly shortened(P <0.05), the immobile swimming time was significantly prolonged(P <0.05),the content of liver sEH protein and mRNA expression levels were significantly increased(P <0.05). Compared with the model group,the total distance of activity and tail hanging struggle time in the fluoxetine hydrochloride group,CLMD-L group and CLMD-H group were significantly extended(P <0.05),the immobile swimming time was significantly shortened(P <0.05),and the liver sEH protein content and mRNA expression levels were significantly decreased(P <0.05). After the intervention,the total distance of activity and tail hanging struggle time in the fluoxetine hydrochloride group,CLMD-L group and CLMD-H group were significantly longer than before the intervention(P <0.05),and the swimming immobility time was significantly shorter than before the intervention(P <0.05). Conclusion The occurrence of depression can increase the expression of sEH in the liver,and Chaihu Jia Longgu Muli Decoction can significantly improve the depression-like behavior and reduce the expression of sEH in the liver of depressed rats. These results indicate that the liver plays an essential role in depression,which may be involved in the occurrence of depression through sEH.