| The energy metabolism of cardiocytes is material basis in heart. The key process is themetabolism of ATP. Mitochondrion is the key organelle about energy metabolism in cells.Therespiratory chain of mitochondrial inner membrane is composed of coenzymes.The electronwas transmited to O2by electron transport chain.In the hole process,H2Oand ATP wereproduced.The major coenzymes in respiratory chain were ComplexⅠ,Ⅱ,Ⅲand Ⅳ.TheComplexⅠwere composed with45subunits that encoded by mitochondrial genes andkaryogene. NADH dehydrogenase subunit4(ND4)is the most important constituent ofComplexⅠ. Evidence shows that the lack of ND4protein have little effect about the structureof ComplexⅠ. Inverse,the activity of ComplexⅠwere lost completely, the activity ofComplexⅡwere lost by50%，the activity of Complex Ⅳwere lost to64%.Furthermore, ithinder the assemble process of ComplexⅠ.So,the ND4protein was very important in energymetabolism of cardiocytes.In this experiment,the ND4protein was research and the relatedmechanism of action were discussed.Objective:Observe the effection of Salidroside to the hypoxia cardiocytes and study themechanism of action with the ND4protein.Methods: First, get the hearts out of20healthy rats that bore in24hours. Digest thehearts with trypsase and obtain cardiocytes in vitro. Culture cells in6and96wells plates.Start the experiment when the cells spread and contact with each others.the cells were dividedinto four groups, control group, model group, hypoxia＋salidroside(50μg/mL)and hypoxia＋salidroside(100μg/mL). The mode design, GENbox anaer, Anaero Pack and low glucoseculture medium with no FBS.The indexs determined were cellular morphology, vitality ofcells, LDH of liquid supernatant, activity of complexⅠ, the change of DNA and cell nucleus,the content of ND4protein.Results:(1)Observed cells under microscope, compared with control group, hypoxia＋salidroside(100μg/mL) group have few apoptosis cells（P＞0.05）and the apoptosis cells weremost in model group（P＜0.01）.(2) The cytoactive were detected by MTT.Result show thatthe cell’s activity in model group were lower than control group cells（P＜0.01）.The level ofcytoactive rose up after adding salidroside with different content.(3)The level of LDH in model group rose up observably compared with control group（P＜0.01）, the level of LDHdescend after adding salidroside with different content.(4)The activity of complexⅠwas leastand in control group was mos（tP＜0.01）.(5)In gel electrophoresis,DNA ladders were emergein model group and hypoxia＋salidroside(50μg/mL) group. On the contrary,there were littleDNA ladders in control group and hypoxia＋salidroside(100μg/mL) group.(6)Observed cellsunder microscope, compared with control group,there were too much apoptosis cells in modelgroup（P＜0.01）.(7) The content of ND4protein in model group was least and in controlgroup was most(P＜0.01).Conclusion:The model of hypoxia was built successfully by using GENboxanaer,Anaero Pack and low glucose culture medium with no FBS.In model group,the level ofLDH,cell’s cytoactive, the activity of complex Ⅰ, and contentof ND4protein were changedcompared with control group.The salidroside have relations with the change.it can decreasethe the level of LDH in hypoxia conditions and rise up the cell’s cytoactive, the activity ofcomplexⅠ,and contentof ND4protein.we guess that salidroside can relieve the injure inhypoxia by protecting mtDNA.What’s more, the structure and function of the electrontransport chain of mitochondria could regular.May be by this way, the salidroside protect thecardiocytes avoid of injuring in hypoxia conditions.We will keep on studing the correlationmachinism later. |
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