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Na ~ / H ~ exchanger inhibitor in cultured neonatal rat cardiomyocytes during hypoxia / reoxygenation injury protection effect and mechanism of

Author: XiaoBin
Tutor: HuangJianQun
School: China Medical University
Course: Emergency Medicine
Keywords: Calcium NHE blockers Hypoxia - reoxygenation injury Cultured neonatal rat myocardial cells
CLC: R541
Type: Master's thesis
Year: 2003
Downloads: 93
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Abstract

Introduction Na ^{/ H ^{exchanger (NHE) is a H ^{discharge cells are transferred to Na ^{of ion channels in ischemia - reperfusion (I / R) injury, especially in myocardial ischemia and early reperfusion, NHE activity leading to intracellular calcium overload, leading to further cell damage important responsibility. According to some reports in the literature, NHE blockers inhibit the activity of NHE, can effectively reduce I / R injury, play a powerful role in myocardial protection, such as the reduction of infarct size, antiarrhythmic, improve endothelial function and reduce apoptosis . But the NHE blockers myocardial protective mechanisms is not yet clear; especially in the application of reperfusion period NHE blockers can play the role of myocardial protection is still controversial, this study using cultured neonatal rat myocardial cells to hypoxia / reoxygenation to simulate myocardial I / R injury by observing a single myocardial cells in hypoxia / cytoplasmic Ca ^{2 concentration (ie the [Ca ^{2 ] < sub> i ) the dynamic changes, and EIPA medication impact on the Ca ^{2 concentration in a different time, while observing the changes of the cell area and cell viability, in order to clarify the the EIPA protection myocardial cells, reducing I / R injury mechanism of action. Materials and Methods 1. Materials 1.1 Animal Wistar rat neonatal rat born 1-2d 1.2 Drugs, 5 - (N-ethyl-N-isopropyl)-amiloride (EI-PA) 2. The method 2.1 I / R injury model was prepared by using a mixed nitrogen P5% nitrogen, 10% carbon dioxide) saturated 30min Tyrode Tyrode's solution (no sugar) replacement bottle culture medium to achieve the purpose of hypoxia, the of observation 180 min. The reoxygenation change DMEM mixed air saturated containing 5% air 5% CO 2) is% fetal bovine serum, the purpose of the I / R injury of observed 70 min 2.2 Experimental groups doors) control group * ONTROL group of people in the lack of reoxygenation, no drug treatment administered p) hypoxia group (EIPA-I group of people in addition to hypoxia of Tyrode's solution and reoxygenation in DMEM uniform contains a 14moUL of the EIPA, more than with the control group. c) before reoxygenation administration group OipA-R; group X at the beginning of the hypoxia non-drug treatment, and 5 min before the start of the end of hypoxia and reoxygenation plus EIPA (advance min dosing so that the drug can give full play to its pharmacodynamics final concentration of lpmoUL, the reoxygenation applications in DMEM containing lpmoUL EIPA K) reoxygenation before the administration of the two groups (EIPA-R group of people the EIPA-R; group EIPA concentration increased to 4pmoVL head. 3. OUTCOME MEASURES the 3.l myocardial cells [Ca Bu; Determination: Load Fur-myocardial cells were placed on an inverted fluorescence microscope to detect The calcium fluorescence excitation wavelength 340nln/380nm emission wavelength of 490-slonm fluorescence image acquisition by the high sensitivity cooled CCD camera digital video camera. fluorescence signal Meta-luo-specific image data analysis software to handle. * az * According to Gynkiewicz, provided the formula. 3.2 Single Cell Size Determination: application the Meta-flllo analysis software for image processing, the cap lock select cells can be obtained by the area of ??the cell. 3.3 myocardial cell survival was statistically Treatment: all test data to X t S said. SPSS.0 computer software system for basic calculation, analysis of variance, linear regression analysis. to Pd.05 as statistically meaningful. · 2 · experimental results. lack of oxygen complex process the EIPA myocardial cells Ka Bu \-R group showed a significant difference between the during reoxygenation, EIPAJ group and the EIPA-RZ group Caz *; rise is much smaller than that of the control group, during which the difference was statistically significant, the EIPA-R; compared with the control There was no significant difference. lack of complex oxygen process of myocardial cell area and the effects of change and EIPA: contracture of myocardial cells of the control group after 30 min of hypoxia, the area was significantly reduced, compared with EIPAJ group significant difference in delay to 90min after area of ??EIPAJ cells was significantly reduced myocardial cells on the the cell contracture comparison complex whole process of oxygen lack, EIPAJ group at the end of reoxygenation cells shrink K7.63 f12.98) The EIPA-R; group (57,63 S 8.16)%, EIPA-R. Group (5 (dish D.27)%, compared with the control group, the door 78. 11.38)% there was a significant difference. 3. The EIPA on cell viability: the survival of the EIPA-I group 4.44%) and the control group and MPA-R; group (M.44% and 50.00% 8 significantly improved, with the EIPA-R Group u1 Stone 2%) no significant difference. discussion myocardial ischemia / reperfusion injury is a complex series of reactions, The mechanism of injury most scholars believe is due to intracellular calcium overload and oxygen free radicals generation and cell adhesion molecules involved in the cause. intracellular calcium overload has been considered to be irreversible cell damage the development of the common pathway in ischemia / reperfusion injury, voltage-dependent channels open, Na * Caz exchanger (NBS) and NHE activity to participate in intracellular calcium overload. the JHE blockers, according to some animal studies have many myocardial protection for, such as reducing infarct size, anti-myocardial stunning. suppression to reduce the role of apoptosis, and so on, but the protection mechanism is not clear; NHE blockers for ischemic reperfusion can achieve the purpose of myocardial protection, becoming the focus of attention of many scholars the study ischemia-reperfusion injury in cultured neonatal rat cardiomyocytes experimental model, the EIPA intervention of the cardiomyocytes during hypoxia and reoxygenation before, the results show that the m factory plant in full of hypoxia / reoxygenation be significantly reduced by the end of the hypoxia and reoxygenation at each time point the concentration of Ca Bu, as well as the increase in the magnitude of hypoxia and reoxygenation Ca * concentration, effectively prevent intracellular calcium super}}}}}}}

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