法舒地尔保护阿霉素对心肌损害的实验研究

发布时间：2018-09-16 19:48

【摘要】：目的:研究阿霉素联合法舒地尔在白血病患者化疗过程中对K562细胞的杀伤作用以及对人心肌细胞的保护作用。方法:选取对数期生长的K562细胞及HCM细胞,分别对其进行药物干预,设立对照组A1及A2,单用阿霉素组B1及B2,阿霉素联合法舒地尔组C1及C2,阿霉素联合右丙亚胺组D1及D2(阳性对照组),干预后的细胞分别在倒置显微镜下通过检测细胞的形态,MTT比色法检测细胞活性,流式检测细胞凋亡率,western blot检测细胞中抑制凋亡基因Bcl-2和促凋亡基因Bax的表达情况。结果:K562细胞实验的结果为:倒置显微镜下与对照组A1相比,实验组B1、C1、D1细胞拒染实验均为阳性;MTT比色法测细胞活性示与对照组A1相比,实验组B1、C1、D1细胞活性均降低,且通过吸光度散点图可以看出三组实验组K562细胞吸光度相近;流式检测细胞凋亡率示与对照组A1((?)=4.93%)相比,实验组B1((?)=81.9%)、C1((?)=78.3%)、D1((?)=86.0%)细胞凋亡率明显升高,且三组间凋亡率差异不大;western条带图示对照组A1及实验组B1、C1、D1的内参条带β-Actin强度基本相等,抑凋亡基因Bcl-2条带强度在对照组A1强度较高,在三组实验组B1、C1、D1条带强度较A1组低,促凋亡基因Bax的条带强度在对照组A1第,在三组实验组B1、C1、D1条带强度较A1组高。HCM细胞实验结果为:倒置显微镜下与对照组A2相比,实验组B2、C2、D2细胞拒染实验均为阳性,且镜下C2、D2组细胞形态较B2组细胞形态变化较少;MTT比色法测细胞活性示与对照组A2相比,实验组B2、C2、D2细胞活性均降低,且通过吸光度散点图可以看出C2、D2组HCM细胞吸光度相近并高于B2组;流式检测细胞凋亡率示与对照组A1((?)=0.13%)相比,实验组B1((?)=42.5%)、C1((?)=24.1%)、D1((?)=23.7%)细胞凋亡率明显升高,且C2、D2组间凋亡率相近并低于B2组;western条带图示对照组A2及实验组B2、C2、D2的内参条带β-Actin强度基本相等,抑凋亡基因Bcl-2条带强度在对照组A1强度最高,其次为C2及D2组,在B2组条带强度最低,促凋亡基因Bax的条带强度在对照组A2最低,在三组实验组B2、C2、D2条带强度较A2增高,C2及D2组条带较高,B2组条带强度最高。结论:在白血病患者化疗过程中,阿霉素联合法舒地尔无明显药物拮抗作用,联合用药在不降低化疗效果的基础上对心肌细胞有一定的保护作用。
[Abstract]:Aim: to study the cytotoxicity of doxorubicin combined with fasudil on K562 cells and its protective effect on human cardiomyocytes during chemotherapy. Methods: K562 cells and HCM cells grew in logarithmic phase were treated with drugs respectively. Control group A1 and A2, adriamycin group B1 and B2, adriamycin combined with fasudil group C1 and C2, adriamycin combined with dextromine group D1 and D2 (positive control group) were used to detect the cells under inverted microscope. The cell activity was detected by MTT colorimetric assay. Apoptosis rate was detected by flow cytometry and western blot was used to detect the expression of Bcl-2 and Bax. Results the results of the cell experiment were as follows: compared with the control group A1 under inverted microscope, the cell activity of the experimental group was decreased by MTT colorimetric assay, while that of the control group was decreased by MTT colorimetric assay compared with that of the control group, and the results showed that the activity of the cells in the experimental group was lower than that in the control group, and that in the control group was significantly lower than that in the control group. The results showed that the absorbance of K562 cells in the three experimental groups was similar to that in the control group, and the apoptotic rate of K562 cells in the experimental group was significantly higher than that in the control group A1 (?) 4.93%), B _ 1 (?) 81.9%), C _ 1 (?) 78.3%) and D _ 1 (?) 86.0%), compared with the control group (A _ 1 (?) 4.93%). There was no significant difference in apoptosis rate among the three groups. The intensity of 尾 -Actin bands in the control group A1 and the experimental group B1C1C1nD1 was basically equal. The intensity of the Bcl-2 band of anti-apoptotic gene was higher in the control group than that in the control group, and the intensity of the B1C1C1mD1 band in the three experimental groups was lower than that in the A1 group. The band intensity of apoptosis-promoting gene Bax was higher in control group A1 than that in A1 group. The results were as follows: compared with control group A _ 2 under inverted microscope, B _ 2C _ 2T _ 2 cells in experimental group were positive in staining rejection test. Under microscope, the changes of cell morphology in group C _ 2 were less than those in group B _ 2. MTT assay showed that the activity of B _ 2 C _ 2O _ 2 cells in the experimental group was lower than that in the control group (A _ 2). The results showed that the absorbance of HCM cells in C _ 2O _ 2 group was similar and higher than that in B _ 2 group, and the apoptotic rate was significantly higher in experimental group B1 (?) 42.5%) than that in control group A1 (?) 0.13%), C _ 1 (?) 24.1%) and D _ 1 (?) 23.7%) in the experimental group, and the apoptosis rate was significantly higher in the C _ 2O _ 2 group than in the control group (A _ 1 (?) 0.13%). The intensity of 尾 -Actin in B _ 2 and B _ 2C _ 2N _ 2 of the control group and the experimental group was basically equal. The intensity of the Bcl-2 band of anti-apoptotic gene was the highest in control group A1, followed by C2 and D2 groups. In B2 group, the band intensity of apoptotic gene Bax was the lowest, and the intensity of B _ 2 C _ 2C _ 2 D _ 2 band in three experimental groups was higher than that of A _ 2 and C _ 2 and D _ 2 groups, and the intensity of B _ 2 band in B _ 2 group was higher than that in B _ 2 group and B _ 2 group was higher than that in D _ 2 group. Conclusion: doxorubicin combined with fasudil has no obvious antagonistic effect during chemotherapy in leukemia patients. The combination of adriamycin and fasudil has a protective effect on cardiomyocytes on the basis of not reducing the effect of chemotherapy.
【学位授予单位】：西安医学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R733.7

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