鹿茸多肽对冈田酸诱导的小鼠海马神经元HT22细胞损伤模型中PI3K、AKT、Caspase-9表达的影响

目的观察鹿茸多肽对冈田酸(OA)诱导的小鼠海马神经元HT22细胞损伤模型中磷脂酰肌醇-3激酶(PI3K)、蛋白激酶B(AKT)、半胱氨酸蛋白酶-9(Caspase-9)表达的影响,探讨鹿茸多肽对HT22细胞损伤模型的保护作用机制。方法采用含10%胎牛血清(FBS)培养液(DMEM/F12)传代培养HT22细胞7d后,分为正常对照组、二甲基亚砜(DMSO)对照组、OA细胞损伤模型组、鹿茸多肽高、中、低剂量组。正常对照组给予含10%FBS的DMEM/F12,DMSO对照组给予DMSO终浓度<0.01%的DMEM/F12,OA细胞损伤模型组给予10nmol OA的DMEM/F12,鹿茸多肽高、中、低剂量组分别给予50、500、1000μg/ml的DMEM/F12,于37℃、5%CO2条件下孵育24h。利用噻唑蓝(MTT)比色法检测细胞存活率,酶联免疫法(ELISA)检测各组实验细胞内PI3K、AKT含量,蛋白质印迹法(Western Blot)检测各组实验细胞内PI3K、AKT、Caspase-9表达水平。结果MTT比色法检测结果表明,与OA细胞损伤模型组比较,鹿茸多肽能够明显提高细胞存活率(P<0.05);ELISA检测结果表明,与OA细胞损伤模型组比较,鹿茸多肽能够明显提高受损HT22细胞内PI3K、AKT含量(P<0.05或P<0.01);Western Blot检测结果表明,与OA细胞损伤模型组比较,鹿茸多肽能够明显提高受损HT22细胞内PI3K、AKT、Caspase-9表达水平(P<0.05或P<0.01)。结论鹿茸多肽对OA诱导的HT22细胞损伤模型具有保护作用,作用机制可能与调节受损HT22细胞内PI3K、AKT、Caspase-9表达水平相关。

Effects of Pilose Antler Polypeptide on Expression of PI3K,AKT,Caspase-9 in Mouse Hippocampal Neuron HT22 Cell Damage Model Induced by Okadaic Acid

Objective Observation of pilose antler peptide on okadaic acid(OA)-induced mouse hippocampal neuron HT22 cell injury model of phosphatidylinositol-3 kinase(PI3K),protein kinase B(AKT),and cysteine proteinase-9(Caspase-9)effect of expression,and explore the protective mechanism of pilose antler peptide on HT22 cell injury model.Methods HT22 cells were subcultured with 10%fetal bovine serum(FBS)medium(DMEM/F12)for 7 days and divided into normal control group,dimethyl sulfoxide(DMSO)control group,OA cell injury model group,and velvet antler.High,medium and low doses of peptides.The normal control group was given DMEM/F12 with 10%FBS,the DMSO control group was given DMEM/F12 with a final DMSO concentration of<0.01%,and the OA cell injury model group was given 10 nmol of OA DMEM/F12.The antler peptide high,medium and low dose groups DMEM/F12 was administered at 50,500,and 1000μg/ml,and incubated at 37℃and 5%CO2 for 24 hours.The cell viability was measured by thiazole blue(MTT)colorimetry,the content of PI3K and AKT in each group of experimental cells was detected by enzyme-linked immunosorbent assay(ELISA),and the PI3K,AKT,and Caspase-9 in each group of experimental cells were detected by Western Blot expression level.Results The MTT colorimetric test results showed that compared with the OA cell damage model group,the velvet antler sample can significantly improve the cell survival rate(P<0.05);the ELISA test results showed that the velvet antler sample can significantly increase the damage compared with the OA cell damage model group.Contents of PI3K and AKT in HT22 cells(P<0.05 or P<0.01);Western blot results showed that velvet antler could significantly increase the expression levels of PI3K,AKT,Caspase-9 in HT22 cells compared with the OA cell damage model group(P<0.05 or P<0.01).Conclusion Antler peptides have a protective effect on OA-induced HT22 cell injury models,and the mechanism may be related to the regulation of PI3K,AKT,and Caspase-9 expression levels in damaged HT22 cells.