鹿茸多肽对冈田酸致大鼠HT22细胞损伤保护作用的实验研究

Protective effect of HT22 cell injury in rats induced by pilose antler polypeptides on Okada acid

为观察鹿茸多肽对冈田酸(Okdaic acid,OA)致大鼠HT22细胞损伤的保护作用,采用硫堇染色法对培养的HT22细胞进行染色鉴定。HT22细胞用含10%FCS的DMEM完全培养液传代培养7d后,分正常对照组、二甲基亚砜(DMSO)对照组、OA损伤模型组和鹿茸多肽3个组共6个组进行条件培养。各组分别加不同的培养液,正常对照组只加含FCS的DMEM;DMSO对照组加含DMSO(终浓度<0.01%)的DMEM;OA损伤模型组加含DMSO(终浓度<0.01%)、终浓度为10nmol/L OA的DMEM;鹿茸多肽3个组分别加含OA(10nmol/L)和鹿茸多肽(终质量浓度分别为50、500、1 000μg/mL)的DMEM,于37℃,5%CO_2孵育24h。通过倒置显微镜观察细胞形态,采用噻唑蓝(MTT)比色法检测细胞存活率,酶联免疫法测定乳酸脱氢酶(lactate dehydrogenase,LDH)渗漏率,采用试剂盒检测细胞上清液中神经递质的变化。结果表明,鹿茸多肽可明显改善HT22的细胞形态,提高细胞的存活率,降低细胞LDH的漏出率,增加神经递质的表达。鹿茸多肽对冈田酸致大鼠HT22细胞损伤具有明显的保护作用。

To observe the protection effect of HT22 cell injury in rats induced by pilose antler polypeptides on Okada acid （OA）, HT22 cells were dyed and identified by sulfur violet staining method. After HT22 cells were which contains 10% FCS complete medium, then they were divided mto six groups to subcultugroup , DMSO control , OA damage modll group and three pilose antler polypeptides groups. The six groups were added different medium. The normal control group only was added DMEM with FCS. The DMSO control group was added DMEM with DMSO （its final concentration lower 0. 01%）. OA damage model group was added DMSO （its final DMEM with OA （its final concentration was 10 nnom/L）. Three pilose antler polypeptides groups were added OA （10 nmol/L） and DMEM with different concentration of pilose antler polypeptides （its final concentrations were 50, 500, 1 000 ^.g/mL） , and they were incubated for 24 hours at 37 C , 5% CO2 . Cell morphology was detected by inverted microscope. Cell viability was de-tected by MTT colorimetric assay. The leakage rate of lactate dehydrogenase （LDH） was determined by ELISA kit The changes of neurotransmitter in cell supernatant were detected by ELISA kit The results show that velvet antler polypeptides can signit-cantly improve the HT22 cell morphology and cell survival rate and reduce the leakage rate of LDH and accelerate the expression of neurotransmitter. Pilose antler peptids has obvious protective effect in rat HT22 cells which injury by Okada acid.