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目的:探讨尖吻蝮蛇小分子多肽对人神经胶质瘤细胞系U251的增殖抑制作用及其机制。方法:MTT法检测2.5、5.0和10.0mg/L尖吻蝮蛇小分子多肽对人神经胶质瘤细胞系U251的增殖抑制作用;RT-PCR法检测尖吻蝮蛇小分子多肽对人神经胶质瘤细胞bcl-2和bax基因表达的影响;分光光度法检测胶质瘤细胞中丙二醛(MDA)的含量和超氧化物歧化酶(SOD)的活性。结果:2.5、5.0和10.0 mg/L尖吻蝮蛇小分子多肽对人神经胶质瘤细胞生长抑制率(49.77%、67.65%和76.42%)高于对照组(P<0.001)。当小分子多肽的剂量增加时,bcl-2 mRNA的表达呈逐渐下降趋势,而bax mRNA的表达则呈逐渐升高趋势,bcl-2/bax亦逐渐减小。2.5、5.0和10.0 mg/L组MDA含量高于对照组(P<0.01),且SOD活性低于对照组(P<0.01);5.0和10.0 mg/L组MDA含量高于2.5mg/L组和对照组(P<0.001),10.0 mg/L组SOD活性低于2.5mg/L组和对照组(P<0.001);10.0 mg/L组SOD活性低于5.0、2.5mg/L组和对照组(P<0.01)。结论:尖吻蝮蛇小分子多肽通过氧化应激使bcl-2/bax mRNA表达降低可能是抑制人神经胶质瘤细胞系U251增殖的重要原因之一。
Objective: To investigate the inhibitory effect of Agkistrodon acutus venom polypeptide on human glioma cell line U251 and its mechanism. Methods: MTT assay was used to detect the inhibitory effects of 2.5, 5.0 and 10.0 mg / L Agkistrodon acutus venom peptides on human glioma cell line U251; RT- And the expression of bcl-2 and bax genes in glioma cells. The content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) in glioma cells were detected by spectrophotometry. Results: The inhibitory rates of growth inhibition of human glioma cells (2.5%, 5.0% and 10.0%) were significantly higher than that of the control group (49.77%, 67.65% and 76.42%, P <0.001). When the dose of small molecule polypeptide increased, the expression of bcl-2 mRNA gradually decreased, while the expression of bax mRNA increased gradually and bcl-2 / bax decreased gradually. The content of MDA in 2.5, 5.0 and 10.0 mg / L groups was higher than that in control group (P <0.01), and the activity of SOD was lower than that in control group (P <0.01) (P <0.001). The SOD activity in 10.0 mg / L group was lower than that in 2.5 mg / L group and control group (P <0.001). The SOD activity in 10.0 mg / L group was lower than that in 5.0 and 2.5 mg / L group Group (P <0.01). Conclusions: The decrease of bcl-2 / bax mRNA expression induced by oxidative stress may be one of the important reasons for inhibiting the proliferation of human glioma cell line U251.