远志寡糖酯对β淀粉样蛋白25~35诱导的SH-SY5Y神经细胞损伤及AKT / CREB / BDNF 信号通路的影响  

Effects of oligosaccharide esters of polygala tenuifolion on β-Amyloid protein 25~35 induced SH-SY5Y cell injury and AKT/CREB/BDNF signal pathway

作　　者：牛芬溪 桑旭星 杨依 唐晓丽 刘雅楠 方芳[1] NIU Fenxi;SANG Xuxing;YANG Yi;TANG Xiaoli;LIU Yanan;FANG Fang(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488,China)

机构地区：[1]北京中医药大学中药学院,北京102488

出　　处：《北京中医药大学学报》2022年第4期414-420,共7页Journal of Beijing University of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(No.81373995)。

摘　　要：目的 观察远志寡糖酯对β淀粉样蛋白25~35(Aβ;)诱导的人成神经母细胞瘤SH-SY5Y细胞损伤及蛋白激酶B/环磷酸腺苷反应元件结合蛋白/脑源性神经营养因子(AKT/CREB/BDNF)信号通路关键凋亡蛋白的影响。方法 将培养的SH-SY5Y细胞分为对照组、Aβ;损伤模型组、远志寡糖酯(50、100、200、400 mg/L)组和Aβ;损伤+远志寡糖酯(50、100、200、400 mg/L)组。对照组和远志寡糖酯组分别给予等体积完全培养液和不同浓度的远志寡糖酯溶液后与细胞共孵育24 h;Aβ;损伤模型组和Aβ;损伤+远志寡糖酯组分别给予等体积完全培养液和不同浓度的远志寡糖酯溶液2 h后,加入25μmol/L Aβ;的溶液与细胞共孵育24 h。采用MTT法检测SH-SY5Y细胞存活率;Hoechst 33258染色观察细胞凋亡;Western Blot检测SH-SY5Y细胞内pAKT/AKT、pCREB/CREB和BDNF蛋白表达。结果 与对照组比较,不同浓度远志寡糖酯(100、200、400 mg/L)能明显增加细胞存活率;Aβ;可降低SH-SY5Y细胞存活率,增强SH-SY5Y细胞内的Hoechst 33258染色荧光强度,降低pAKT/AKT、pCREB/CREB和BDNF的蛋白表达(P<0.01)。与Aβ;损伤模型组比较,不同浓度的远志寡糖酯(100、200、400 mg/L)均可明显提高细胞存活率,降低细胞内Hoechst 33258染色的荧光强度,减少细胞凋亡,增加细胞内BDNF、 pAKT/AKT、pCREB/CREB的表达(P<0.05,P<0.01)。结论 远志寡糖酯抗Aβ;诱导的SH-SY5Y神经细胞损伤可能与激活AKT/CREB磷酸化,促进BDNF表达有关。Objective To investigate the effect of oligosaccharide esters(OE) of polygala tenuifolion(Yuanzhi) on β-Amyloid protein(Aβ;) induced SH-SY5 Y cell injury and AKT/CREB/BDNF signal pathway. Methods The cultured SH-SY5 Y cells were divided into control group, Aβ;injury model group, OE(50, 100, 200 and 400 mg/L) groups, and Aβ;injury + OE(50, 100, 200 and 400 mg/L) group. Cells in the control group and OE group were incubated with equal volume of solvent and different concentrations of OE 24 hours, respectively. Cells in the Aβ;injury model group and Aβ;injury + OE group were incubated with 25 μmol/L Aβ;for 24 hours after the addition of equal volume of solvent and different concentrations of OE for 2 h. The survival rate of SH-SY5 Y cells was detected by MTT assay;Hoechst 33258 staining was used to observe apoptosis;The expressions of pAkt/Akt, pCREB/CREB and BDNF proteins in SH-SY5 Y cells were detected by Western blot. Results Compared with the control group, OE(100, 200 and 400 mg/L) significantly increased the survival rate of SH-SY5 Y cells;Aβ;significantly decreased the survival rate of SH-SY5 Y cells, increased the fluorescence intensity of Hoechst 33258 staining, and decreased the protein expression of pAkt/Akt, pCREB/CREB and BDNF in cells(P<0.01). Compared with the model group, OE(100, 200 and 400 mg/L) treatment significantly enhanced the cell survival rate, decreased the fluorescence intensity of Hoechst 33258 staining, reduced apoptosis, and increased the expression of BDNF, pAkt/Akt, pCREB/CREB in cells(P<0.05, P<0.01). Conclusion OE has protective effects against β-Amyloid protein induced SH-SY5 Y cell injury, which is related to activating Akt/CREB phosphorylation and promoting the expression of BDNF.

关 键 词：远志寡糖酯 β淀粉样蛋白25~35 人成神经母细胞瘤SH-SY5Y细胞 脑源性神经营养因子 

分 类 号：R285.5[医药卫生—中药学]