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Comparison of Heptanol Precondictioning, Hypioxic Preconditioning and Hypoxic Postconditiong by Cx43 of Myocardial Mitochondria on Anoxia/Reoxygenation Injury in Rat Myocardium Cells

Author: LiWeiKe
Tutor: ZhongGuoQiang
School: Guangxi Medical University
Course: Department of Cardiology
Keywords: Heptanol Myocardial cells Mitochondria Cx43 Hypoxia / reoxygenation Hypoxia preconditioning After hypoxia treatment Mitochondrial Cx43
CLC: R541.4
Type: Master's thesis
Year: 2011
Downloads: 45
Quote: 0
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Abstract


Today's prevention role in ischemia-reperfusion (I / R) injury has become an important issue to be solved in coronary atherosclerotic heart disease treatment, the study found that the gap junctional communication involved in the I / R injury. Gap junctions as a target for anti-ischemic reperfusion injury may be a new research direction of myocardial protection. Heptanol (Heptanol) as gap junction blockers against I / R injury protection mechanism is not clear, especially the mitochondrial Cx43 mediated myocardial hypoxia / reoxygenation injury protective effect has not been reported in the literature. Therefore, the subject is the establishment of myocardial hypoxia / reoxygenation model (AR) applications heptanol pretreatment detect mitochondrial Cx43 content of its cell apoptosis and ischemic preconditioning and post-processing on myocardial cells lack oxygen / reoxygenation injury in myocardial mitochondrial Cx43. The purpose of research through the cell membrane and mitochondrial Cx43 expression changes, explore different concentrations of heptanol (Heptanol) pretreatment on myocardial cells hypoxia / reoxygenation injury mechanisms at the subcellular level. Primary myocardial cell cultures born 1-3 days SD rats were randomly divided into 7 groups: normal control group (control group), hypoxia / reoxygenation group (AR group), plasmin group (DMSO group) , 0.1mmol / L heptanol preconditioning group (0.1HT group), 0.5 mmol / L heptanol preconditioning group (0.5HT group), 1.0 mmol / L heptanol preconditioning group (1.0HT group), 2.0 mmol / L heptyl the alcohol pretreatment group (2.0HT group). Control group, hypoxia / reoxygenation and HT drug intervention. AR group, the cultured myocardial cells during hypoxia / reoxygenation for 30 minutes, and then incubated for 120 minutes in the incubator under normal conditions; the DMSO group of myocardial cell culture flasks and HT group equal amount of DMSO carried hypoxia heptanol pretreatment / reoxygenation treatment; other groups to join the appropriate concentration (0.1 mmol / L and 0.5mmol / L and 1.0mmol / L and 2.0mmol / L), and then be hypoxia / reoxygenation. RT-PCR technology from the level of transcription detect Cx43mRNA expression levels change. Extraction of mitochondria, mitochondrial Cx43 protein content detected by Western blot. AR group compared the control group Cx43mRNA expression levels and mitochondrial Cx43 content decreased significantly (P lt; 0.01). 1.0 HT group, 2.0HT AR group comparison, Cx43mRNA expression levels and mitochondrial Cx43 content was significantly increased (P lt; 0.01). 0.1HT group and 0.5 HT group compared with AR group, Cx43mRNA expression levels and mitochondrial Cx43 content was no significant difference (P gt; 0.05). Conclusion heptanol pretreatment AR lead to cell membranes and mitochondrial Cx43 reduction can be reversed, but the effect concentration. Mitochondrial Cx43 may be involved in the cardioprotective effect of heptanol pretreatment. Heptanol purpose at the subcellular level pre-processing, processing ischemic preconditioning and ischemic rat heart mitochondrial Cx43 mediated hypoxia / reoxygenation injury mechanisms. Primary myocardial cell cultures, born 1-3 days SD rats were randomly divided into six groups: normal control group (control group), hypoxia / reoxygenation group (AR group), plasmin group (DMSO group) , hypoxia preconditioning group (HP group) after hypoxia treatment group (PC group), heptanol preconditioning group (HT group). Control group, hypoxia / reoxygenation and HT drug intervention. AR group, the cultured myocardial cells during hypoxia / reoxygenation pretreatment 30 minutes, and then replace the normal culture medium and incubated for 120 minutes in the incubator under normal conditions; DMSO group in myocardial cell culture flasks with HT group amount of DMSO solvent, hypoxia / reoxygenation treatment; HP group, to be hypoxic preconditioning of cardiac myocytes, hypoxia / reoxygenation treatment; PC group first be hypoxia / reoxygenation, and then be hypoxia treatment ; HT group 1.0mmol / L heptanol in culture flasks, and then be hypoxia / reoxygenation. Cell apoptosis was detected by flow cytometry. RT-PCR technology from the level of transcription detect Cx43mRNA expression levels change. Extraction of mitochondria, mitochondrial Cx43 protein content detected by Western blot. Results AR group, DMSO group compared to the control group apoptosis rate increased significantly (P lt; 0.01), Cx43mRNA expression levels and mitochondrial Cx43 content was significantly reduced (P lt; 0.01). HP group, the PC group and the HT group and AR group compared cardiomyocyte apoptosis rate decreases (P lt; 0.01) Cx43mRNA expression levels and mitochondrial Cx43 content increased significantly (P lt; 0.01). HP group, the PC group compared to the HT group apoptosis rate, Cx43mRNA expression levels and mitochondrial Cx43 content was no significant difference (P gt; 0.05). Conclusion heptanol protective effect of preconditioning on hypoxia / reoxygenation its protective effect may be achieved by affecting the expression of cardiac mitochondria Cx43 with of hypoxia pretreatment and after treatment similar.

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CLC: > Medicine, health > Internal Medicine > Heart, blood vessels ( circulatory ) disease > Heart disease > Coronary arteries ( atherosclerosis ),heart disease (CHD)
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