Shi Bo-Yu, Rao Zhi-Li, Luo Jie, Liu Xiao-Bo, Fang Yang, Cao Hai-Juan, Zeng Nan
Pharmacy College,Chengdu University of Traditional Chinese Medicine Chengdu 611137,China.
Zhongguo Zhong Yao Za Zhi. 2019 Feb;44(4):781-786. doi: 10.19540/j.cnki.cjcmm.20181101.003.
To investigate the relationship between anti-depressant effect and hippocampal nerve growth of Xiaoyao San,the inflammatory model of hippocampal neuron was induced by lipopolysaccharide( LPS). The effect of Xiaoyao San serum( final concentration of4%,8%) on the cell proliferation activity was detected by immunofluorescence,the levels of BDNF and β-NGF in the supernatant of hippocampal neurons were detected by ELISA,and the expressions of BDNF,NGF,Trk B,Trk A and CREB mRNA in cell lysate of hippocampal neuron were detected by PCR. Western blot was used to detect the expressions of Trk B,CREB,p-CREB and SYP protein in cell lysate of hippocampal neuron,and to reveal the neuroprotective effect and mechanism of Xiaoyao San. The results showed that8% Xiaoyao San serum could significantly increase in Brdu/Neu N ratio( P<0. 01). 4%,8% Xiaoyao San serum could significantly improve the levels of BDNF and β-NGF in supernatant( P<0. 05 or P<0. 01),up-regulate the expression of BDNF,NGF,Trk B,Trk A,CREB mRNA and Trk B,p-CREB,SYP protein in cell lysate( P< 0. 05 or P< 0. 01). 8% Xiaoyao San serum could significantly increase CREB protein in cell lysate( P<0. 05),and elevate in p-CREB/CREB ratio( P<0. 01). All the above results indicate that Xiaoyao San has a certain protective effect on LPS induced hippocampal neuron injury,which suggests that the protective effect of Xiaoyao San is related to the promotion of hippocampal nerve growth,which is one of its antidepressant mechanisms.
为探讨逍遥散抗抑郁作用与海马神经生长之间的关系，采用脂多糖（LPS）诱导海马神经元炎症模型。通过免疫荧光检测逍遥散含药血清（终浓度4%、8%）对细胞增殖活性的影响，采用ELISA法检测海马神经元培养上清中脑源性神经营养因子（BDNF）和β-神经生长因子（β-NGF）水平，采用PCR法检测海马神经元细胞裂解液中BDNF、NGF、酪氨酸激酶B（Trk B）、酪氨酸激酶A（Trk A）和环磷腺苷效应元件结合蛋白（CREB）mRNA的表达。采用Western blot法检测海马神经元细胞裂解液中Trk B、CREB、磷酸化CREB（p-CREB）和突触素（SYP）蛋白的表达，以揭示逍遥散的神经保护作用及机制。结果显示，8%逍遥散含药血清能显著提高5-溴脱氧尿嘧啶核苷（Brdu）/神经元核抗原（Neu N）比值（P<0.01）。4%、8%逍遥散含药血清能显著提高培养上清中BDNF和β-NGF水平（P<0.05或P<0.01），上调细胞裂解液中BDNF、NGF、Trk B、Trk A、CREB mRNA及Trk B、p-CREB、SYP蛋白的表达（P<0.05或P<0.01）。8%逍遥散含药血清能显著增加细胞裂解液中CREB蛋白含量（P<0.05），提高p-CREB/CREB比值（P<0.01）。上述结果表明，逍遥散对LPS诱导的海马神经元损伤具有一定的保护作用，提示逍遥散的保护作用与促进海马神经生长有关，这可能是其抗抑郁作用机制之一。
