Background: Recent studies have demonstrated that pyroptosis has an important impact on the pathological process of many cardiovascular diseases, and inhibiting this process can significantly reduce pathological damag...Background: Recent studies have demonstrated that pyroptosis has an important impact on the pathological process of many cardiovascular diseases, and inhibiting this process can significantly reduce pathological damage. At the same time, some studies have proved that Zhuangtongy in (ZTY) has a certain protective effect on the myocardium of rats with coronary heart disease. Therefore, the aim of this study is to observe and explore the effects of ZTY on cardiomyocytes and its mechanism of action in rats with coronary heart disease based on the Caspase-1 mediated cellular pyroptosis pathway. Methods: Fifty SD rats were randomly divided into blank group, sham-operation group, model group, ZTY group and cysteinyl aspartate specific protease (Caspase-1) inhibitor group. The blank group did not receive any treatment, and the other groups established a rat model of coronary artery disease by ligating the left anterior descending branch of the coronary artery, but the sham-operation group was only threaded without ligation. After that, the sham-operated and model groups were gavaged with 0.1 ml/kg distilled water, the ZTY group was gavaged with 13.6 g/kg ZTY decoction based on the previous study, and the Caspase-1 inhibitor group was injected intraperitoneally with 3 mg/kg Ac-YVAD-cmk solvent once a day for four consecutive weeks. Transmission electron microscopy was used to observe the ultrastructure of cardiomyocytes, HE staining was used to observe the morphology of cardiac tissue, and enzyme-linked immunosorbent assay (Elisa) was used to detect serum interleukin-1β (IL-1β), interleukin-18 (IL-18) and C-reactive protein (CRP) in the rats. In addition, fluorescent probe was used to detect serum Reactive oxygen species (ROS). Real-time fluorescence quantitative PCR was used to detect the expression of mRNAs of ASC, Nlrp3, Caspase-1 and gasdermin-D (GSDMD), and the protein expression of Caspase-1 and GSDMD was detected by immunohistochemistry. Results: There was no significant difference between the results of the blank and sham-operated groups. Compared with the blank group, the transmission electron microscopy results showed swollen and ruptured cardiomyocyte membranes in the model group, with pore formation, severe mitochondrial swelling, membrane lysis, and cristae breakage. The staining results showed myofibril breakage, severe intercellular oedema and vacuolation in the model group. The inflammatory factors IL-1β, IL-18, CRP and ROS were significantly elevated (P β, IL-18, CRP, ROS and the expression of ASC, Nlrp3, Caspase-1, and GSDMD mRNA were significantly elevated (P Conclusion: ZTY can reduce myocardial injury by lowering the expression of ROS and other inflammatory factors, inhibiting Caspase-1 mediated cellular pyroptosis, thus reducing myocardial inflammation and protecting cardiomyocytes.展开更多
文摘Background: Recent studies have demonstrated that pyroptosis has an important impact on the pathological process of many cardiovascular diseases, and inhibiting this process can significantly reduce pathological damage. At the same time, some studies have proved that Zhuangtongy in (ZTY) has a certain protective effect on the myocardium of rats with coronary heart disease. Therefore, the aim of this study is to observe and explore the effects of ZTY on cardiomyocytes and its mechanism of action in rats with coronary heart disease based on the Caspase-1 mediated cellular pyroptosis pathway. Methods: Fifty SD rats were randomly divided into blank group, sham-operation group, model group, ZTY group and cysteinyl aspartate specific protease (Caspase-1) inhibitor group. The blank group did not receive any treatment, and the other groups established a rat model of coronary artery disease by ligating the left anterior descending branch of the coronary artery, but the sham-operation group was only threaded without ligation. After that, the sham-operated and model groups were gavaged with 0.1 ml/kg distilled water, the ZTY group was gavaged with 13.6 g/kg ZTY decoction based on the previous study, and the Caspase-1 inhibitor group was injected intraperitoneally with 3 mg/kg Ac-YVAD-cmk solvent once a day for four consecutive weeks. Transmission electron microscopy was used to observe the ultrastructure of cardiomyocytes, HE staining was used to observe the morphology of cardiac tissue, and enzyme-linked immunosorbent assay (Elisa) was used to detect serum interleukin-1β (IL-1β), interleukin-18 (IL-18) and C-reactive protein (CRP) in the rats. In addition, fluorescent probe was used to detect serum Reactive oxygen species (ROS). Real-time fluorescence quantitative PCR was used to detect the expression of mRNAs of ASC, Nlrp3, Caspase-1 and gasdermin-D (GSDMD), and the protein expression of Caspase-1 and GSDMD was detected by immunohistochemistry. Results: There was no significant difference between the results of the blank and sham-operated groups. Compared with the blank group, the transmission electron microscopy results showed swollen and ruptured cardiomyocyte membranes in the model group, with pore formation, severe mitochondrial swelling, membrane lysis, and cristae breakage. The staining results showed myofibril breakage, severe intercellular oedema and vacuolation in the model group. The inflammatory factors IL-1β, IL-18, CRP and ROS were significantly elevated (P β, IL-18, CRP, ROS and the expression of ASC, Nlrp3, Caspase-1, and GSDMD mRNA were significantly elevated (P Conclusion: ZTY can reduce myocardial injury by lowering the expression of ROS and other inflammatory factors, inhibiting Caspase-1 mediated cellular pyroptosis, thus reducing myocardial inflammation and protecting cardiomyocytes.
