Effect of Xinkang Granule(心康冲剂)on Cardiomyocyte Pyroptosis in Rats with Chronic Heart Failure
Objective To investigate the regulatory effect of Xinkang Granule(心康冲剂)on cardiomyocyte pyroptosis in rats with chronic heart failure(CHF).Methods Six of 46 SPF male and famale SD rats were randomly selected as the blank group,and the other 40 rats were injected with adriamycin intraperitoneally to establish CHF model.After the model was successful,32 rats were randomly divided into model group(n=12),Xinkang Granule group(n=10)and valsartan group(n=10).The valsar-tan group was given 0.03 g/(kg·d)equivalent dose by gavage,and the Xinkang Granule group was given 0.5 g/(kg·d)e-quivalent dose by gavage.The blank group and model group were given the same amount of normal saline daily for 4 weeks.At the end of the experiment,the cardiac function of each group was detected by cardiac ultrasound.HE and Masson staining were used to observe the morphology of heart tissue.The ultrastructure of myocardial tissue was observed under transmission electron microscope(TEM).The expression site of pyroptosis substrate gasdermin D-N(GSDMD-N)was detected by immunohisto-chemistry,and enzyme-linked immunosorbent assay was used to detect the expressions of interleukin-1 β(IL-1 β)and inter-leukin-18(IL-18)in serum of rats.Real-time PCR was used to detect mRNA of NOD-like receptor thermal protein domain associated protein 3(NLRP3)pathway in each group.Western blot was used to detect the protein expression levels of NLRP3 pathway.Results Compared with those of the blank group,left ventricular ejection fractions(LVEF)and left ventricular fractional shortening(LVFS)were significantly decreased(P＜0.01)in the model group.The results of staining showed the increased number of myocardial cell death and the promotion of fibrosis.Moreover,GSDMD-N was expressed in the cell membrane and cytosol.In the model group,the results of TEM showed that the myocardial membrane was broken,the pore formation was ob-served,and the mitochondrial swelling was severe.Inflammatory factors IL-18 and IL-1β in blood were significantly increased(P＜0.01).The expression of NLRP3,Caspase-1,GSDMD mRNA and GSDMD-N protein were significantly increased in my-ocardial tissue(P＜0.01).Compared to the model group,after the intervention of Xinkang Granule and valsartan,cardiomyocyte pyroptosis was reduced,the values of left ventricular ejection fraction(LVEF)and left ventricular fraction shortening(LVFS)were increased(P＜0.01).The results of staining and TEM showed that the pathological damage of myocardium was relieved.The expressions of IL-18,IL-1 β,NLRP3,caspase-1 and GSDMD/GSDMD-N were decreased(P＜0.01).Conclusion Xin-kang Granule can improve the cardiac function of CHF rats,inhibit the NLRP3 pathway,reduce the myocardial inflammation and prevent the excessive myocardial cell death in CHF rats.
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