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EFFECT OF NIMODIPINE ON HL-60 CELL APOPTOSIS AND ITS MECHANISM

Baoyu Gao

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Abstract

ObjectiveTo study the effect of Nimodipine on HL-60 and its mechanism.MethodsThe standard HL-60 cells were divided into control and experimental groups.Nimodipine(0.01 g/L) was added into the experimental group. The dynamic changes were observed through an inverted microscope and morphological changes through a microscope with Giemsa method.The DNA fragments were detected through agarose gel electrophoresis.The content of DNA was analyzed through a flow cytometer.The expression of apoptosis-related genes bcl-2 and bax were investigated using immunohistochemical technique.Results(In experimental) group: from after eight hours of cell culture,the cells showed deforming,bubbling;while the membrane of the cells and bubbles remained integrity.With the extension of the culture time,the number of bubbled cell increased,cell capacity decreased,some cells split;no morphological changes were noted in the control group.With Giemsa staining,the split of cell membrane was seen and the chromatin appeared as purplish red,and concentrated into pieces,approaching to nuclear membrane,gathering to the edge and forming typical apoptosis bodies.DNA ladders on agarose gel were clearly seen at 8 h,16 h and 24 h in the experimental group.The apoptosis rate increased in the experimental group with time elongating,being markedly statistically significant from 8 h compared with the control one.The optical density(OD) values of Bcl-2 protein expression obviously decreased in the experimental group,being different(t=3.66,5.62;P0.05) by 16 h and 24 h compared with the control group.The OD values of Bax obviously elevated,being different(t=4.13,P0.05) by 16 h and significantly different(t=7.55,P0.01) by 24 h compared with the control group.The rate of bcl-2/Bax protein descended and showed a significant difference(t=5.32-7.76,P0.01) by 8 h,16 h and 24 h compared with the control group.ConclusionNimodipine could effectively induce apoptosis of HL-60 cells.The decrease of the proportionality of Bcl-2/Bax protein might be one of the mechanisms inducing apoptosis of HL-60.

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ObjectiveTo study the effect of Nimodipine on HL-60 and its mechanism.MethodsThe standard HL-60 cells were divided into control and experimental groups.Nimodipine(0.01 g/L) was added into the experimental group. The dynamic changes were observed through an inverted microscope and morphological changes through a microscope with Giemsa method.The DNA fragments were detected through agarose gel electrophoresis.The content of DNA was analyzed through a flow cytometer.The expression of apoptosis-related genes bcl-2 and bax were investigated using immunohistochemical technique.Results(In experimental) group: from after eight hours of cell culture,the cells showed deforming,bubbling;while the membrane of the cells and bubbles remained integrity.With the extension of the culture time,the number of bubbled cell increased,cell capacity decreased,some cells split;no morphological changes were noted in the control group.With Giemsa staining,the split of cell membrane was seen and the chromatin appeared as purplish red,and concentrated into pieces,approaching to nuclear membrane,gathering to the edge and forming typical apoptosis bodies.DNA ladders on agarose gel were clearly seen at 8 h,16 h and 24 h in the experimental group.The apoptosis rate increased in the experimental group with time elongating,being markedly statistically significant from 8 h compared with the control one.The optical density(OD) values of Bcl-2 protein expression obviously decreased in the experimental group,being different(t=3.66,5.62;P0.05) by 16 h and 24 h compared with the control group.The OD values of Bax obviously elevated,being different(t=4.13,P0.05) by 16 h and significantly different(t=7.55,P0.01) by 24 h compared with the control group.The rate of bcl-2/Bax protein descended and showed a significant difference(t=5.32-7.76,P0.01) by 8 h,16 h and 24 h compared with the control group.ConclusionNimodipine could effectively induce apoptosis of HL-60 cells.The decrease of the proportionality of Bcl-2/Bax protein might be one of the mechanisms inducing apoptosis of HL-60.

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Available abstract

ObjectiveTo study the effect of Nimodipine on HL-60 and its mechanism.MethodsThe standard HL-60 cells were divided into control and experimental groups.Nimodipine(0.01 g/L) was added into the experimental group. The dynamic changes were observed through an inverted microscope and morphological changes through a microscope with Giemsa method.The DNA fragments were detected through agarose gel electrophoresis.The content of DNA was analyzed through a flow cytometer.The expression of apoptosis-related genes bcl-2 and bax were investigated using immunohistochemical technique.Results(In experimental) group: from after eight hours of cell culture,the cells showed deforming,bubbling;while the membrane of the cells and bubbles remained integrity.With the extension of the culture time,the number of bubbled cell increased,cell capacity decreased,some cells split;no morphological changes were noted in the control group.With Giemsa staining,the split of cell membrane was seen and the chromatin appeared as purplish red,and concentrated into pieces,approaching to nuclear membrane,gathering to the edge and forming typical apoptosis bodies.DNA ladders on agarose gel were clearly seen at 8 h,16 h and 24 h in the experimental group.The apoptosis rate increased in the experimental group with time elongating,being markedly statistically significant from 8 h compared with the control one.The optical density(OD) values of Bcl-2 protein expression obviously decreased in the experimental group,being different(t=3.66,5.62;P0.05) by 16 h and 24 h compared with the control group.The OD values of Bax obviously elevated,being different(t=4.13,P0.05) by 16 h and significantly different(t=7.55,P0.01) by 24 h compared with the control group.The rate of bcl-2/Bax protein descended and showed a significant difference(t=5.32-7.76,P0.01) by 8 h,16 h and 24 h compared with the control group.ConclusionNimodipine could effectively induce apoptosis of HL-60 cells.The decrease of the proportionality of Bcl-2/Bax protein might be one of the mechanisms inducing apoptosis of HL-60.

Key concepts: Apoptosis, Nimodipine, Agarose gel electrophoresis, Molecular biology, Agarose, Giemsa stain, Cell, Gel electrophoresis

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EFFECT OF NIMODIPINE ON HL-60 CELL APOPTOSIS AND ITS MECHANISM — Research Paper | ScholarLens