广州医科大学学报

目的：筛选半夏厚朴汤抗抑郁的活性部位，并初步揭示其抗抑郁的分子机制。方法：半夏厚朴汤提取浓缩后，AB-8大孔树脂柱层析，经梯度乙醇洗脱得到水（W）、30%(30%E)、50%(50%E)和70%乙醇（70%E）部位。强迫游泳实验（FST）、旷场实验（OFT）和脂多糖（LPS）抑郁模型筛选半夏厚朴汤抗抑郁的有效部位。ELISA和实时荧光定量PCR(QPCR)检测小鼠炎性细胞因子表达；蛋白免疫印迹法（Western blotting）测定炎症相关蛋白水平。结果：LPS注射后小鼠表现出抑郁样行为，包括FST中不动时间增加，OFT中运动距离减少；W和70%E部位治疗可逆转LPS引起的抑郁样行为。ELISA和QPCR结果显示：LPS增加小鼠血清、前额叶皮层（PFC）和海马（HPC）中TNF-α、IL-1β、皮质酮（CORT）含量，增加TNF-α、IL-1β mRNA表达；W、70%E部位治疗可以抑制炎性因子的增加。Western blotting显示，W和70%E部位治疗可抑制LPS注射引起的PFC和HPC中AKT、NF-κB p65、TNF-α、JAK2和STAT3蛋白水平增加。结论：半夏厚朴汤发挥抗抑郁作用的活性部位可能是W和70%E组分，可能通过调控AKT/NF-κB/JAK2/STAT3抑制炎症反应发挥抗抑郁作用。

Objective:To explore the extracted fractions of Banxia Houpo Decoction for antidepressant activity and elucidate their potential mechanisms.Methods:After extraction and concentration,Banxia Houpo Decoction was transferred to an AB-8 macroporous resin column. The water(W),30% ethanol(30%E),50% ethanol(50%E),and70% ethanol(70%E) fraction were obtained with gradient ethanol elution. Forced Swim ming Test(FST) and Lipopolysaccharide(LPS)injection induced depression model was used to screened the antidepressant-active fractions.Enzyme-linked immunosorbent assay(ELISA)and quantitative real-time PCR(QPCR)were used to detect the expression of inflammatory cytokines in mice,while Western blotting was employed to deter mine the expression of inflammation-related proteins in PFC and HPC.Results:Mice exhibited depression-like behaviors after LPS injection,characterized by increased immobility time in the FST and reduced locomotor distance in the OFT. Treatment with the W and 70%E fractions reversed these depression-like behaviors induced by LPS. ELISA and QPCR results showed that LPS injection increased TNF-α,IL-1β,corticosterone(CORT)content in serum,prefrontal cortex(PFC),and hippocampus(HPC),and increased the mRNA expression level of TNF-α,IL-1β. Treatment with W and 70%E fraction inhibited the increase in these inflammatory factors. Western blotting analysis revealed that treatment with theW and 70%E fractions suppressed the increased protein levels of AKT,NF-κB p65,TNF-α,JAK2 and STAT3 in thePFC and HPC induced by LPS injection.Conclusion:The active fractions of the antidepressant effect exerted byBanxia Houpo Decoction may be the W and 70% E fractions,which may exert antidepressant effects by inhibitinginflammatory responses through the modulation of the AKT/NF-κB/JAK2/STAT3 signaling pathway.