毛蕊花糖苷对皮质酮损伤HT22细胞保护作用研究

Protective effect of verbascoside on corticosterone injury HT22 cells

目的 探讨毛蕊花糖苷对皮质酮诱导小鼠海马神经元细胞株HT22损伤的保护作用及其机制。方法 用皮质酮构建慢性应激损伤HT22神经元细胞的体外模型。将HT22细胞分为空白组、模型组、雌二醇(E2)组和毛蕊花糖苷组。空白组与模型组以DMEM培养24 h;雌二醇组加入DMEM和0.3μmol·L^(-1)E2溶液;毛蕊花糖苷组加入DMEM和10μmol·L^(-1)毛蕊花糖苷溶液,培养2 h后,除空白组外,其余各组加入400μmol·L^(-1)的皮质酮储备液,继续培养22 h。采用流式细胞术检测各组细胞凋亡率;以蛋白质印迹法检测脑源性神经营养因子(BDNF)、原肌球蛋白受体激酶B(TrkB)、雌激素受体(ERβ)蛋白表达。结果 正常组、模型组、雌二醇组和毛蕊花糖苷组的细胞凋亡率分别为(3.31±0.35)%、(20.82±0.73)%、(9.35±0.87)%和(7.73±0.62)%,BDNF蛋白表达水平分别为1.16±0.02、0.17±0.02、0.70±0.02和0.43±0.03,TrkB蛋白表达水平分别为1.18±2.00×10^(-3)、0.35±3.00×10^(-3)、0.72±0.01和0.52±4.00×10^(-3),ERβ蛋白表达水平分别为1.22±2.00×10^(-3)、0.58±4.00×10^(-3)、0.80±0.01和0.64±4.00×10^(-3)。上述指标,模型组与正常组比较,差异均有统计学意义(均P<0.05);雌二醇组和毛蕊花糖苷组与模型组比较,差异均有统计学意义(均P<0.05)。结论 毛蕊花糖苷可促进皮质酮损伤的HT22细胞增殖,抑制细胞凋亡,具有一定的细胞保护作用,其机制与E2/ERβ/BDNF/TrkB信号通路有关。

Objective To investigate the protective effect of verbascoside oncorticosteronee-induced HT22 injuryiin mouse hippocampal neuronal cell line and its mechanism.Methods Anin vitro model of HT22 neuronal cells damaged by chronic stresswas constructed using corticosterone.HT22 cells were divided into blank group,model group,estradiol(E2)group and verbascoside group.The blank and model groups were incubated with DMEM for 24 h;the estradiol group was incubated with DMEM and O.3μmol·L^(-1) E2 solution;the verbascoside group was incubated with DMEM and 10μmol·L-I verbascoside solution for 2 h.After 2 h,all groups except the blank group were incubated with 400μmol·L-'corticosterone stock solution and continued to incubate for 22 h.The apoptosis rate of each group of cells was detected by flow cytometry;the expression of brain-derived neurotrophic factor(BDNF),tropomyosin receptor kinase B(TrkB),and estrogen receptor(ERβ)was detected by Western blotting.Results The apoptosis levels of normal group,model group,estradiol group and verbascoside group were(3.31±0.35)%,(20.82±0.73)%,(9.35±0.87)%and(7.73±0.62)%,respectively;the expression levels of BDNF protein were 1.16±0.02、0.17±0.02、0.70±0.02 and 0.43±0.03,respectively;TrkB protein expression levels were 1.18±2.00×10^(-3)、0.35±3.00×10^(-3)、0.72±0.01 and 0.52±4.00×10^(-3),respectively;ERβprotein expression levels were 1.22±2.00×10^(-3)、0.58±4.00×10^(-3)0.80±0.01 and 0.64±4.00×10^(-3),respectively.The above indicators:Compared with the normal group,the differences were statistically significant(all P<0.05);both estradiol group and verbascoside group had statistical significance compared with model group(all P<0.05).Conclusions Verbascoside promotes the proliferation of corticosterone-injured HT22 cells,inhibits apoptosis and has a certain cytoprotective effect by a mechanism related to the E,/ERβ/BDNF/TrkB signaling pathway.