The Discovery Of Pyroptosis Of Intestinal Cell And The Exploration Of Biomarkers After Ionizing Radiation

Objective:From the perspective of pyroptosis,we tried to explore the role of this death mode in the small intestine after radiation,and at the same time,tried to find the relevant biomarker molecules,thus providing a basis for judging the severity of intestinal radiation sickness and clinically targeted therapy.Methods:1、The western blot technique detected the expression of caspase-1,caspase-11,caspase-3,GSDMD and GSDME in all small intestine,proximal small intestine,and distal small intestine at 3h,6h,24 h,48h and 72 h after whole body irradiation with 6Gy,8Gy,11 Gy and 13 Gy in mice.2、The contents of IL-1β,IL-18,IL-22,IL-23,and IL-33 in the plasma of mice at3 h and 6h after whole body irradiation with 6Gy,8Gy,11 Gy and 13 Gy were detected by ELISA.3、Dot blot technique was used to detect the expression of REG3 G in plasma of mice at 3h and 6h after whole body irradiation with 6Gy,8Gy,11 Gy and 13 Gy in mice.4、Dot blot technique was used to detect the expression of REG3 G and IL-1β in plasma of radiotherapy patients before and after radiation therapy.5、A biochemical analyzer was used to measure plasma K+,Cl-,Ca2+,Na+,Mg2+and P plasma levels in mice exposed to 15 Gy of abdomen irradiation.6、 The ICP-OES technique was used to detect the outflow of K+ and Cl-after exposure to 2Gy in HCT116 and sw480 cells.Results:1、 The expression of cleaved caspase-1 was decreased and the expression of cleaved caspase-3 was increased within 6h after whole body irradiation with 11 Gy in mice small intestine.2、The expression of cleaved caspase-1 in the proximal small intestine of normal mice could be detected.3、Within 3 hours after 6Gy and 11 Gy irradiation in mice,the expression of cleaved caspase-1 in the proximal small intestine increased with increasing dose,and the expression of GSDMD in the distal small intestine increased with increasing dose.4、Before and after the mice received a certain dose of whole body irradiation,the plasma levels of IL-18 and IL-23 did not change.5、No IL-1β,IL-22,and IL-33 expression was detected in mice within 6 hours after irradiation with 6Gy,8Gy,11 Gy,and 13 Gy.6、The level of REG3 G in plasma was increased within 6 hours after whole-body irradiation of mice receiving 6 Gy,8 Gy,11 Gy,and 13 Gy.7 、 After treatment with radiation therapy,plasma REG3 G levels of patients increased.8、After the mice were irradiated with 15 Gy in the abdomen,the K+and Cl-levels in plasma increased.9、The K+ continued to flow out within 3h after exposure to 2Gy in HCT116 and sw480 cells.Conclusion:1、When ionizing radiation induces pyroptosis in cells,the death mechanisms of cells in the upper and lower parts of the small intestine are different.2、Three hours after 11 Gy whole body irradiation,there was GSDMD-mediated cell pyroptosis in the lower small intestine of mice.3、Cellular K+ extracellular flux may serve as an early marker of radiation-induced cell death.