基于NLRP3/Capase-1通路探讨芪参养心汤对缺氧条件下大鼠心肌细胞的影响OA

Objective To investigate the effect of Qishen Yangxin Decoction and miR-223-3p on rat cardiomyocytes under hypoxic conditions through regulation of the NLRP3/Caspase-1 pathway.Methods SD rats were randomly divided into a blank control group and low-,medium-,and high-dose Qishen Yangxin Decoction groups.The blank control group was gavaged with an equal volume of physiological saline,while the low-,medium-,and high-dose Qishen Yangxin Decoction groups were gavaged with Qishen Yangxin Decoction at doses of 0.9,1.8,and 3.6 g/mL respectively.All groups were administrated once daily for 7 consecutive days.Blood was collected from the abdominal aorta 1 hour after the last administration to obtain serum.The blank serum and Qishen Yangxin Decoction-containing serum at various concentrations were co-cultured with H9C2 cells for 12 hours.Cell proliferation was assessed using the CCK-8 assay to determine the optimal intervention concentration of the drug-containing serum.Then the H9C2 cells were randomly divided into control group,model group,Qishen Yangxin Decoction drug-containing serum group,miR-223-3p mimics group,miR-NC mimics group,miR-223-3p inhibitor group,and miR-NC inhibitor group.Transfection was performed in miR-223-3p mimics group,miR-NC mimics group,miR-223-3p inhibitor group,and miR-NC inhibitor group.After successful transfection,hypoxia modeling were established in the model group,Qishen Yangxin Decoction drug-containing serum group,miR-223-3p mimics group,miR-NC mimics group,miR-223-3p inhibitor,and miR-NC inhibitor group.After successful modeling,the LDH level in the cell supernatant was detected;the content of IL-1β and IL-18 in the supernatant was detected by Elisa method,the expression of NLRP3 was detected by qRT-PCR method;the expression of relevant proteins NLRP3,cleaved Caspase-1,ASC,GSDMD-N,IL-1β,IL-18 and TNF-α in H9C2 cells was determined by Western blot method.Results Compared with the control group,the model group showed significantly increased LDH level in the supernatant,along with significantly increased expression of NLRP3-mRNA and the related proteins NLRP3,cleaved Caspase-1,ASC,GSDMD-N,IL-1β,IL-18,and TNF-α in H9C2 cells(P<0.05).Compared with the model group,the Qishen Yangxin Decoction drug-containing serum group and the miR-223-3p mimics group showed significantly decrease in LDH level in the supernatant,along with significantly decreased expression of NLRP3 mRNA and the related proteins NLRP3,cleaved Caspase-1,ASC,GSDMD-N,IL-1β,IL-18,and TNF-α in H9C2 cells(P<0.05).Compared with the Qishen Yangxin Decoction drug-containing serum group and miR-223-3p mimics group,miR-223-3p inhibitor group showed significantly increase LDH level in the supernatant,along with significantly increased expression of NLRP3 mRNA and the related proteins NLRP3,cleaved Caspase-1,ASC,GSDMD-N,IL-1β,IL-18,and TNF-α in H9C2 cells(P<0.05).Conclusion Qishen Yangxin Decoction can significantly inhibit the pyroptosis of H9C2 cells in hypoxia modeling of heart failure.Its mechanism may involve targeting the NLRP3/Caspase-1 pathway.Meanwhile,miR-223 is also involved in the expression of the NLRP3/Caspase-1 pathway,thereby influencing the progression of heart failure.