Objective To investigate the effect of palmitoleic acid(POA)on pyroptosis of cardiomyocytes after hy-poxia/reoxygenation-induced injury in the human myocardium.Methods The experiment comprised a control group(Control,normal culture),a hypoxia/reoxygenation group(HR),a palmitoleic acid-treated group(HR+POA),and an anhydrous ethanol control group(HR+ET).Cardiomyocytes viability was assessed using CCK-8 assay,and the level of pyroptosis of cardiomyocytes was measured through the double staining with Hoechst33342/PI and LDH assay.ELISA was employed to detect the release of inflammatory factors IL-1 β and IL-18 in the cell culture supernatant.qRT-PCR and Western blot were utilized to determine the relative expression levels of mRNA and protein of pyroptosis-related genes,namely NLRP3,ASC,Caspase-1,GSDMD,IL-1 β and IL-18,respective-ly.Results CCK-8 assay showed that the survival of hypoxic/reoxygenated cardiomyocytes increased with the ad-dition of POA at concentrations ranging from 25 to 100 μmol/L,as compared to the HR group;a hypoxia/reoxy-genation model of cardiomyocyte was established.The expression of protein and mRNA increased in NLRP3,ASC,Cleaved caspase-1,GSDMD-N,IL-Iβ and IL-18 vs the control group(P＜0.05),the positive percentage of Ho-echst33342/PI staining in cardiomyocytes increased significantly(P＜0.05),the release of LDH,IL-Iβ,and IL-18 increased(P＜0.05).After intervention with 100 μmol/L POA,the protein and mRNA expression levels of NLRP3,ASC,Cleaved caspase-1,GSDMD-N,IL-Iβ,and IL-18 were significantly reduced in the HR+POA group vs HR+ET group(P＜0.05).The positive percentage of Hoechst33342/PI staining in cardiomyocytes de-creased significantly,and the levels of LDH,IL-Iβ and IL-18 significantly decreased(P＜0.05).Conclusion Palmitoleic acid may alleviate hypoxia/reoxygenation-induced injury of cardiomyocytes by inhibiting pyroptosis and inflammatory response after hypoxia/reoxygenation in human myocardium.

In order to explore whether the expression of CircFoxO3 is related to the proliferation and apoptosis of porcine myoblasts,this study transfected porcine myoblasts with CircFoxO3 over-expression vector and siRNA,and detected cell proliferation and apoptosis by EdU and Annexin V-mCherry fluorescence staining,and detected the expression of apoptosis-related genes by RT-qPCR.The results showed that after transfection with overexpression vector,the proportion of ap-optotic cells increased and the number of cells proliferation decreased.At the same time,the ex-pression of apoptosis-related genes Caspase-3,Caspase-8,Fas and FasL was significantly up-regula-ted by RT-qPCR(P＜0.05),and the expression of Bax was extremely significantly up-regulated(P＜0.01).The expression of Bcl-2 was significantly down-regulated(P＜0.05).After transfection with si-CircFoxO3,the apoptosis level decreased and proliferation ability increased.the expression of Bax and FasL genes was highly significantly down-regulated(P＜0.01),the expression of caspase-3 and Caspase-8 was significantly decreased(P＜0.05),and the expression of Bcl-2 was significantly increased(P＜0.05).CircFoxO3 expression inhibited proliferation of porcine myo-blasts,promoted the expression of apoptosis-related genes,and promoted apoptosis of myoblasts.

To investigate the regulatory effect of thioredoxin-interacting protein(TXNIP)on the proliferation and apoptosis of bovine mammary epithelial cells,a bovine TXNIP gene overexpres-sion vector was constructed using the homologous recombination method.The overexpressed re-combinant plasmid was verified through enzyme digestion and sequencing,and the obtained se-quenccs were subjected to BLAST analysis and phylogenetic construction.The TXNIP overcxpres-sion plasmid was then transfected into bovine mammary epithelial cells.Cell proliferation was as-sessed using CCK-8 and EdU assays,while apoptosis was evaluated using Annexin V-mCherry.Ad-ditionally,RT-qPCR was employed to measure changes in the mRNA expression of apoptosis-re-lated genes in bovine mammary epithelial cells.The results showed that the CDS region of bovine TXNIP gene was 1 176 bp,encoded 391 amino acids,which was closely related to sheep.Compared to the control group,overexpression of the TXNIP gene decreased the viability of mammary epi-thelial cells,reduced cell proliferation,and promoted apoptosis.Specifically,overexpression of the TXNIP gene led to a significant upregulation of apoptosis-related genes,including Caspasc-3,Caspase-9,and Bax,in bovine mammary epithelial cells(P＜0.05).In contrast,the expression of Caspase-8 had no significant changes(P＞0.05).Additionally,the expression of anti-apoptosis gene Bcl-2 was significantly decreased(P＜0.01),and the ratio of Bax/Bcl-2 was significantly increased(P＜0.01).In conclusion,the TXNIP gene can inhibit the proliferation of bovine mam-mary epithelial cells,promote apoptosis,and regulate the expression of apoptosis-related genes,thereby influencing the development of the bovine mammary gland.

In order to explore whether the expression of CircFoxO3 is related to the proliferation and apoptosis of porcine myoblasts,this study transfected porcine myoblasts with CircFoxO3 over-expression vector and siRNA,and detected cell proliferation and apoptosis by EdU and Annexin V-mCherry fluorescence staining,and detected the expression of apoptosis-related genes by RT-qPCR.The results showed that after transfection with overexpression vector,the proportion of ap-optotic cells increased and the number of cells proliferation decreased.At the same time,the ex-pression of apoptosis-related genes Caspase-3,Caspase-8,Fas and FasL was significantly up-regula-ted by RT-qPCR(P＜0.05),and the expression of Bax was extremely significantly up-regulated(P＜0.01).The expression of Bcl-2 was significantly down-regulated(P＜0.05).After transfection with si-CircFoxO3,the apoptosis level decreased and proliferation ability increased.the expression of Bax and FasL genes was highly significantly down-regulated(P＜0.01),the expression of caspase-3 and Caspase-8 was significantly decreased(P＜0.05),and the expression of Bcl-2 was significantly increased(P＜0.05).CircFoxO3 expression inhibited proliferation of porcine myo-blasts,promoted the expression of apoptosis-related genes,and promoted apoptosis of myoblasts.

To investigate the regulatory effect of thioredoxin-interacting protein(TXNIP)on the proliferation and apoptosis of bovine mammary epithelial cells,a bovine TXNIP gene overexpres-sion vector was constructed using the homologous recombination method.The overexpressed re-combinant plasmid was verified through enzyme digestion and sequencing,and the obtained se-quenccs were subjected to BLAST analysis and phylogenetic construction.The TXNIP overcxpres-sion plasmid was then transfected into bovine mammary epithelial cells.Cell proliferation was as-sessed using CCK-8 and EdU assays,while apoptosis was evaluated using Annexin V-mCherry.Ad-ditionally,RT-qPCR was employed to measure changes in the mRNA expression of apoptosis-re-lated genes in bovine mammary epithelial cells.The results showed that the CDS region of bovine TXNIP gene was 1 176 bp,encoded 391 amino acids,which was closely related to sheep.Compared to the control group,overexpression of the TXNIP gene decreased the viability of mammary epi-thelial cells,reduced cell proliferation,and promoted apoptosis.Specifically,overexpression of the TXNIP gene led to a significant upregulation of apoptosis-related genes,including Caspasc-3,Caspase-9,and Bax,in bovine mammary epithelial cells(P＜0.05).In contrast,the expression of Caspase-8 had no significant changes(P＞0.05).Additionally,the expression of anti-apoptosis gene Bcl-2 was significantly decreased(P＜0.01),and the ratio of Bax/Bcl-2 was significantly increased(P＜0.01).In conclusion,the TXNIP gene can inhibit the proliferation of bovine mam-mary epithelial cells,promote apoptosis,and regulate the expression of apoptosis-related genes,thereby influencing the development of the bovine mammary gland.