Exploring The Association Of Cell Pyroptosis Regulated By TLR4,NF-κB And NLRP3 And Radiation Cardiac Injury

Objective: To construct a mouse model of radiation heart injury with different doses,observe the pathological changes,determine the expression levels of TLR4,NF-κB,NLRP 3,IL-6,IL-I0,IL-1β,and TNF-α,and investigate the relationship between the activation of TLR4/NF-κB pathway by NLRP3 inflammasome and radiation heart injury.Methods: A mouse model of radioactive heart injury was established,and 30 C57 BL / 6 mice(females)raised under the same conditions were randomly divided into 3 groups of 10 mice:control group,experiment 1 and experiment 2.In the control group,no treatment,the experimental group was irradiated with biological X-ray irradiated at different doses(dose rate100 c Gy / min),1 cm 1 cm.The irradiation dose was 2 Gy in experiment 1 and 20 Gy in experiment 2.Mice were killed 26 weeks after a single irradiation,myocardial tissue was taken for pathological section staining,and morphological changes of fibroblasts were observed in rat myocytes,fibroblasts and cells with hematoxylin-eosin(HE)staining.The expression levels of inflammatory cytokines IL-6,IL-10,IL-1 β and TNF-α m RNA were measured by real-time polymerase chain reaction(QPCR),and the relative expression levels of TLR 4,NF-κB and NLRP 3 were measured by protein immunoimprinting(Western blot).Results: 1.Normal cardiomyocytes are long arrangement,arranged neatly,the nucleus is located in the center of the cell,oval,light dye,blue purple,cytoplasm in red.Fibroblasts are spindle-shaped with long nuclei in the center,dark blue,and cytoplasm in red and intact vascular architecture.At 26 weeks after irradiation,cardiomyocytes were disorganized with enlarged spaces,mild pyknotic nuclei,nuclear staining enlargement,more fibroblasts and partial breaks.Was most obvious in the group with 20 Gy irradiation.2.Comparison with the control group: The expression levels of IL-1 β,IL-6,IL-10,and TNF-α m RNA were significantly increased in the myocardial tissue of each irradiated group at 26 weeks after radiation irradiation,and the difference was statistically significant(P < 0.05).Compared with the 2 Gy group: the expression levels of IL-6,TNF-α and IL-10 m RNA were significantly increased in the 20 Gy group,and the differences were statistically significant(P < 0.05);however,there was no signif-cant difference in IL-1 β m RNA expression between the two irradiation groups,not statistically significant(P > 0.05).3.Compared with the control group,the relative expression levels of TLR4,NF-κB and NLRP 3 were significantly increased in the myocardial tissues of mice irradiated 26 weeks after radiation irradiation,with the 20 Gy group(P < 0.05).Comparison between the two irradiation groups,the expression levels of TLR4 and NF-κB in the 20 Gy group were statistically significant(P < 0.05);there was no significant difference in NLRP3 between the two irradiation groups(P > 0.05).Conclusion: 1.Different doses of single radiation irradiation changed the morphology of cardiomyocytes.The higher the dose,the more obvious,indicating that the severity of cardiac injury is related to the dose.2.Radial irradiation may cause the expression of TLR 4,NF-κB,NLRP 3,IL-6,IL-10,IL-1 β,and TNF-α,which may participate in the occurrence and development of radioactive cardiac injury.pyroptosis mediated by TLR4,NF-κB,and NLPR3 is associated with radioactive cardiac injury;the degree of pyroptosis and inflammatory response in mouse cardiac tissues after different doses.