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目的研究人参皂苷Rg1对β-淀粉样肽(Aβ25-35)诱导的神经母细胞瘤细胞(SK-N-SH细胞)损伤的保护作用。方法用Aβ25-35处理人SK-N-SH细胞,模拟Alzhermer′s(AD)病人体内神经元病理损伤,并以人参皂苷Rg1拮抗其作用。采用MTT比色法检测细胞活力;应用流式细胞仪检测细胞凋亡率;DNA梯型观察凋亡细胞的DNA片段化;化学比色法测定细胞培养液和细胞内丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性;RT-PCR检测细胞淀粉样肽蛋白前体(APP)基因、中性内肽酶(NEP)基因的表达情况;Western Blot检测细胞APP和NEP蛋白的表达情况。结果20μmol/LAβ25-35诱导SK-N-SH细胞24 h细胞活力下降(均P<0.01),细胞凋亡率为33.9%,DNA断裂呈片段状,细胞培养液及细胞内MDA含量增加,SOD活性降低。而人参皂苷Rg1能提高细胞的存活率,减少细胞损伤,使细胞培养液及细胞内MDA含量降低,SOD活性提高(P<0.01),RT-PCR结果显示人参皂苷Rg1可降低APP基因的表达并提高NEP基因的表达,同时Western Blot结果显示人参皂苷Rg1可降低APP蛋白的表达并提高NEP蛋白的表达。结论人参皂苷Rg1对Aβ25-35造成的细胞毒性具有一定的保护作用,其作用机理可能与人参皂苷Rg1能提高SK-N-SH细胞的抗氧化能力以及下调APP基因的表达,上调NEP基因的表达,从而抑制细胞凋亡有关。
Objective To study the protective effect of ginsenoside Rg1 on neuronal blastoma cells (SK-N-SH cells) induced by β-amyloid peptide (Aβ25-35). Methods Human SK-N-SH cells were treated with Aβ25-35 to simulate neuronal pathological injury in patients with Alzhermer’s (AD) and antagonized its effects with ginsenoside Rg1. MTT colorimetric assay was used to detect cell viability; flow cytometry was used to detect the apoptosis rate; DNA ladder was used to observe the DNA fragmentation of apoptotic cells; and chemical colorimetry was used to determine cell culture medium and intracellular malondialdehyde (MDA) content. And superoxide dismutase (SOD) activity; RT-PCR detection of cell amyloid peptide protein (APP) gene, neutral endopeptidase (NEP) gene expression; Western Blot detection of cell APP and NEP protein expression Happening. Results The cell viability of SK-N-SH cells induced by 20μmol/L Aβ25-35 was decreased (all P<0.01), the apoptosis rate was 33.9%, DNA fragmentation was fragmented, cell culture medium and intracellular MDA content increased, SOD Reduced activity. The ginsenoside Rg1 can increase the survival rate of cells, reduce cell damage, reduce cell MDA content and intracellular MDA content (SOD activity) (P<0.01), RT-PCR results show that ginsenoside Rg1 can reduce the expression of APP gene and The expression of NEP gene was increased, and Western Blot showed that ginsenoside Rg1 could reduce the expression of APP protein and increase the expression of NEP protein. Conclusion Ginsenoside Rg1 has a protective effect on cytotoxicity induced by Aβ25-35. Its mechanism of action may be related to that ginsenoside Rg1 can increase the antioxidant capacity of SK-N-SH cells, down-regulate the expression of APP gene and up-regulate the expression of NEP gene. This inhibits apoptosis.