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Objective To investigate the effect and mechanism of homocysteine on apoptosis and proliferation of cardiomyocytes. Methods Cardiomyocytes were stimulated with homocysteine at the concentration of 3 mmol/L for 24 h. Cells were divided into control group and homocysteine group (Hcy group). Western blotting was detected the expression of TLR4,Myeloid differentiation factor 88 (MyD88), NF-κB, p-NF-κB, the expression of apoptosis related proteins Bax, cleaved-caspase-3 and anti-apoptotic protein Bcl-2; Flow cytometry was detected the apoptosis rate of cardiomyocyte, EdU staining was observed the proliferation of cardiomyocytes. After TLR4 interference fragment was transfected into cardiomyocytes, the interference fragment was verified by Western blotting. The cells were transfected with si-NC or si-TLR4 and treated with Hcy. Western blotting was detected the expression of MyD88, NF-κB and p-NF-κB, the expression of apoptosis related protein and anti-apoptosis protein in cardiomyocytes; The apoptosis rate of cardiomyocytes was detected by flow cytometry; The apoptosis of cardiomyocytes was observed by EdU staining. Results Compare with control group, Western blotting showed that TLR4, MyD88 and p-NF-κB significantly increased in Hcy group (P<0.01); The expression of apoptosis related proteins Bax and cleaved-caspase-3 increased and the expression of anti-apoptotic protein Bcl-2 was decreased (P<0.01); Compared with si NC+Hcy group, the expression of TLR4, MyD88 and p-NF-κB decreased (P<0.01), the expression of apoptosis related protein decreased (P<0.01), and the expression of anti-apoptosis protein increased in si-TLR4+Hcy group (P<0.01); Compare with control group, the results of flow cytometry showed that the apoptosis rate of cardiomyocytes increased in Hcy group (P<0.01); Compared with si-NC+Hcy group, the apoptosis rate of si-TLR4+Hcy group decreased (P<0.01). Compared with control group, EdU staining showed that the number of positive cells decreased in Hcy group (P<0.01); Compared with si-NC+Hcy group, the number of positive cells increased in si-TLR4+Hcy group (P<0.01). Conclusion Hcy can induce apoptosis and inhibit proliferation of cardiomyocytes, the mechanism may have something to do with TLR4/NF-κB signaling pathway.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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