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目的研究阿里红不同极性萃取物对β-淀粉样蛋白25-35片段(Aβ25-35)诱导大鼠肾上腺嗜络瘤细胞(PC12)损伤的保护作用。方法阿里红不同极性萃取物作用于PC12细胞,四甲基偶氮唑蓝(MTT)法筛选出对PC12细胞增殖活性最强的萃取物,Aβ25-35诱导PC12细胞损伤建立阿尔茨海默病体外模型;将培养的细胞分为5组:正常对照组、模型组(Aβ25-35损伤组)、药物低剂量组(50μg/mL)、药物中剂量组(100μg/mL)、药物高剂量组(200μg/mL)。采用MTT法检测PC12细胞的存活率;AinexinV-FITC/7-AAD双染法流式细胞仪检测细胞凋亡;采用RT-PCR法检测bax和bcl-2基因mRNA的表达水平。结果在阿里红不同极性萃取物中,乙酸乙酯萃取物可促进PC12细胞的增殖,差异具有统计学意义(P<0.05);给予Aβ25-35诱导后PC12细胞存活率降低为(44.73±10.14)%,细胞凋亡率升高(19.6%);而给予阿里红乙酸乙酯萃取物处理后,高剂量组细胞存活率升高(64.36±4.00)%,细胞凋亡率降低(6.1%),阿里红乙酸乙酯萃取物高、中、低剂量组的细胞存活率差异有统计学意义。RTPCR结果表明经Aβ25-35处理后细胞的bax基因表达上调,bcl-2基因表达下调;给予阿里红乙酸乙酯萃取物处理后,bax基因表达水平明显降低,bcl-2基因表达水平明显增高。结论阿里红不同极性萃取物中乙酸乙酯萃取物对Aβ25-35诱导PC-12细胞损伤有保护作用。
Objective To study the protective effects of different polar extracts of Alison on the injury of adrenal sinuses tumor cells (PC12) induced by β-amyloid 25-35 fragment (Aβ25-35). Methods Extracts of different polarities were treated in PC12 cells. Methyl thiazolyl tetrazolium (MTT) method was used to extract the extract with the strongest proliferative activity on PC12 cells. Aβ25-35 induced PC12 cell injury and established Alzheimer’s disease In vitro, the cultured cells were divided into 5 groups: normal control group, model group (Aβ25-35 injury group), low dose group (50μg / mL), medium dose group (100μg / mL) (200 μg / mL). The survival rate of PC12 cells was detected by MTT assay. Apoptosis was detected by flow cytometry with Ainexin V-FITC / 7-AAD double staining. The mRNA expression of bax and bcl-2 was detected by RT-PCR. Results Ethyl acetate extract could promote the proliferation of PC12 cells in different polar extracts of Aliex, the difference was statistically significant (P <0.05); the survival rate of PC12 cells induced by Aβ25-35 was decreased to (44.73 ± 10.14 ), The apoptotic rate increased (19.6%). After being treated with ethyl acetate extract of Aliri, the cell survival rate increased (64.36 ± 4.00)% and the cell apoptosis rate decreased by 6.1% There was significant difference in cell viability between high, middle and low doses of ethyl acetate extract of Alibaba. The results of RTPCR indicated that the expression of bax gene was up-regulated and the expression of bcl-2 gene was down-regulated after treatment with Aβ25-35. The expression of bax gene and the expression of bcl-2 gene were significantly increased after the treatment with ethidium acetate extract. CONCLUSION: Acetate extracts from extracts of different polarity have protective effects on Aβ25-35-induced PC-12 cell injury.