Myocardial Cells In Studies Of Autophagy And Traditional Chinese Medicines

Autophagy which occurred in eukaryotic cells from lysosomes degradate the initial content is an important physiological processes in the body stability, biological synthesis, cell development, organizational remodeling, environmental adaptation, disease prevention, anti-aging and so on can not be ignored significance. In recent years, since the autophagy with huge potential arising from research and application of medical, biological widespread attention in academic circles, but the mechanism has not yet clear, the role of traditional Chinese medicine research also not been reported.In the literature of the subject, under the guidance of SD rat cardiac cells for the experiment to immunofluorescence staining technology group and the immune Western blot technique to study means to autophagy specific proteins such as Beclin1 and Micro-associated protein 1 light chain 3 for the inspection targets, around the autophagy model, the impact of drugs on the autophagy, autophagy cell damage associated with the three parts of content, depth layers of the myocardial cells and autophagy since the role of traditional Chinese medicine.Some of the research are summarized as follows:The first part of myocardial cells and macrophages model autophagy time-related researchThis study aimed at the establishment of viable cells autophagy research model, and to explore the strength and macrophages induced autophagy time.The original culture of rat cardiac cells, the establishment of nutritious environment training control group (CM) and in the designated time window built into the sugar-free serum-free (glucose deprive, GD) Environmental Training autophagic of the five model : GD3h, GD6h, GD12h, GD18h, GD24h group;A group of immunofluorescence staining and Western Blot analysis technology group since the macrophage-specific expression of differences, the results are as follows:1. GD model than the CM Group, since the macrophage-specific protein Beclin1, LC3II expression increased, while the rate of increase LC3I relatively small, LC3II/LC3I increased tips GD environment since the bubble increase in the number of macrophages, the cells were induced autophagy .2. Beclin1 among all groups of expression and LC3II/LC3I change with the trends for GD3h Group> GD6h Group> GD24h Group> GD12h Group> GD18h Group> CM Group, namely autophagy intensity sort.3. Autophagy-induced autophagy initial stage of a strong, with the induction time extension, the first since the macrophages strength gradually decreased and then be enhanced.The results showed that:1. Myocardial cells autophagy model successfully established.2. Autophagy intensity and duration of a non-linear GD related.3. Note: Since the macrophage cells respond to cell damage is enhanced as a response to speculation since the macrophage cells in damage and repair in the process of playing an important role.4. On the basis of this select autophagy strong GD3h, GD6h, GD24h model for the next study. The second part of ginsenoside Rg1, Rb1, Re on myocardial cells on the role of autophagyExperimental 1 ginsenoside Rg1, Rb1, Re concentration on the administrationThe experiments are designed to study ginsenoside Rg1, Rb1, Re myocardial cells of rat dynamic impact, guiding the administration of experimental settings.The original culture of rat cardiac cells, were given the concentration 0.1,1,10,100,1000 mol / L of ginsenoside Rg1, Rb1, Re of 24 h, MTT Determination of the group OD value, inspection and control of drug-treated Inter-group differences.The results showed that: Only concentration 1000 mol / L of ginsenoside Rg1, Re OD value of the treatment group and control group there were significant differences (P <0.01). The results showed that: In addition to ginsenoside Rg1, Re concentration in 1000 mol / L above the myocardial cell injury heavier, the remainder of the drug concentration on the impact of myocardial viability was not obvious.Reference to the literature of drugs commonly used, the experiment to determine ginsenoside Rg1, Rb1, Re 100 mol / L, 10 mol / L as a high or low dose, the drug study of myocardial cells from the impact of macrophages.Experiment 2 ginsenoside Rg1, Rb1, Re on myocardial cells from the impact of macrophagesThis experiment to study drugs on the role of autophagy cells, macrophages and the establishment of cells and cell damage since the relevance of the research base.Experimental divided into the normal control group (CM Group); GD3h, GD6h, GD24h model group and its corresponding level of synchronization dosage group to study ginsenoside Rg1, Rb1, Re set in the concentration of 100 mol / L, 10 mol / L of the myocardial Since the impact of macrophage cells.A group of immunofluorescence staining and Western Blot analysis technology group since the macrophage-specific expression of differences, the results are as follows: GD compared with the model group, treatment group Beclin 1 expression was significantly decreased, or with the dose was positively correlated. GD compared with the model group, treatment group LC3I relatively small changes, LC3II expression and LC3II/LC3I decreased significantly, or with the dose was positively correlated. The results showed that: ginsenoside Rg1, Rb1, Re to inhibit cell autophagy, intensity and dose was positively related to the mechanism and stop the autophagy bubble formation.On this basis, the cell autophagy cell damage associated with the study.The third part of autophagy cell injury and the relevance of the studyThe experiments are designed to study the model and the corresponding GD synchronized delivery of the Group of injury, since the study cells and macrophages damage relations.Experimental division takes a reference of Experiment 2. Collection of all groups cell culture medium, the concentration of LDH.The results showed that: 1. Compared with the control group, the model group LDH concentration increased significantly (P <0.01).2. The model for the sort of LDH concentration GD24h Group> GD6h Group> GD3h group.3. The same time window treatment group compared with the model group, LDH concentration decreased to GD6h, GD24h high and low dose treatment group significantly (P <0.01).4. The same time window between the administration group, LDH lower dose rate and was positively correlated.The results showed that:1. In selected within the time window autophagy in intensity and extent of damage was negatively correlated.2. Ginsenoside Rg1, Rb1, Re effective in inhibiting the release of LDH at the same time, inhibit autophagy occurred two drugs inhibit both concentration and dependence.3. Ginsenoside Rg1, Rb1, Re on myocardial cells have protective effects, its protection mechanisms and the promotion of autophagy has nothing to do.The experiment has not yet been involved the autophagy-specific regulation of research, no critical judgement of the relationship between cell damage and autophagy can be infer, just speculate (1) within the selected time window,cell autophagy protection and injury at the same time there, and the protect role is more critical. (2) Ginsenoside Rg1, Rb1, Re confrontational autophagy in regulating the cell damage.