Effect and mechanism of Shenyuan Yiqi Huoxui Capsules on TNF-α-induced myocardial cell injury in mice
Objective To investigate the effect of Shenyuan Yiqi Huoxue Capsules(SYYQ)on tumor necrosis factor α(TNF-α)-induced myocardial cell injury in mice and its mechanism.Methods Mouse cardiomyocytes(HL-1 cells)were cultured routinely,and SYYQ-containing serum was prepared.HL-1 cells were treated with 0%,10%,20%,and 50%SYYQ-containing serum.Cell viability was measured using the cell counting kit-8(CCK-8)assay,and the optimal experimental concentration was selected based on the cell survival rate.HL-1 cells were randomly divided into three groups:the control group,which was cultured with DMEM;the model group,which was stimulated with 50 ng/mL TNF-α for 12 h to induce myocardial injury;the SYYQ group,which was treated with the optimal concentration of drug-containing serum for 12 h.Flow cytometry was used to detect apoptosis,and real-time PCR was used to measure the mRNA expression levels of nuclear factor kappa B(NF-κB),nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3(NLRP3),and interleukin-18(IL-18).Western blotting was used to detect the protein expression levels of NF-κB,NLRP3,and IL-18.Results The survival rates of myocardial cells were(53.32±5.70)%,(68.24±6.41)%,(92.62±8.56)%,and(86.93±7.12)%,respectively.The 20%drug-containing serum group was selected as the optimal concentration for drug intervention.The apoptosis rates of cardiomyocytes in the control group,model group,and SYYQ group were(1.57±0.18)%,(18.99±1.32)%,and(6.76±0.38)%,respectively.Compared with the control group,the apoptosis rate in the model group was significantly higher(P<0.01).Compared with the model group,the apoptosis rate in the SYYQ group was significantly lower(P<0.01).The mRNA expression levels of NF-κB,NLRP3,and IL-18 were significantly higher in the model group compared with the control group(P<0.05).Compared with the model group,the mRNA expression levels of NF-κB,NLRP3,and IL-18 were significantly lower in the SYYQ group(P<0.01).The relative protein expression levels of NLRP3 and IL-18 were significantly higher in the model group compared with the control group(P<0.01).Compared with the model group,the relative protein levels of NF-κB,NLRP3,and IL-18 were significantly lower in the SYYQ group(P<0.05,P<0.01).Conclusion SYYQ may reduce TNF-α-induced myocardial cell injury by regulating the NF-κB/NLRP3/IL-18 signaling pathway.
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