The Effect Of Amiloride On PI3K/AKT Signaling Pathway In ARDS Rats With Lung Injury

Purpose Through the oleic acid-induced ARDS rat model,the effect of amiloride on acute lung injury is explored and whether this effect is achieved through the PI3 K / Akt signaling pathway.Methods 96 male Wistar rats were randomly divided into blank control group,DMSO group,ARDS model group,and amiloride intervention group.The ARDS rat model induced by oleic acid was established.Collect the serum and bronchoalveolar lavage fluid of rats at 4 h,8 h and 12 h of injection oleic acid,and the wet weight(LW)and dry weight(DW)of rat lung tissue were weighed,and calculated LW / DW ratio and rat lung water clearance(AFC);Elisa method was used to detect uPAR,TNF-?,IL-6 and IL-10 levels in rat serum and bronchoalveolar lavage fluid;HE staining method was used to observe lung tissue disease in rats physiological changes;immunohistochemical methods were used to detect the positive expression levels of VEGF,VCAM-1 and ET-1 in rat lung tissues;RT-PCR was used to detect uPAR,PI3 K,Akt and GSK 3? mRNA expression levels;Western blot was used to detect uPAR,PI3 K,p-PI3 K,Akt,p-Akt and GSK 3? protein expression levels in rat lung tissues.Results(1)After injection of oleic acid for 4 h,8 h and 12 h,compared with the blank control group,the LW / DW was significantly increased,and the AFC value was significantly decreased in the ARDS group.After the intervention of amiloride,the LW / DW decreased significantly,and the AFC value increased significantly;(2)Elisa results showed that uPAR,TNF-?,and IL-6 levels in serum and bronchoalveolar lavage fluid in ARDS rats increased significantly,and IL-10 levels decreased significantly.After amiloride intervention,the contents of uPAR,TNF-? and IL-6 in the serum and bronchoalveolar lavage fluid of the rats were significantly reduced,and the levels of IL-10 were significantly increased.(3)The results of HE staining showed that the alveolar wall of the lung tissue of the ARDS group was thickened,the alveolar structure was destroyed,and the inflammatory cells were infiltrated;amiloril intervention could improve this pathological change.(4)The results of immunohistochemistry showed that the positive expression of VEGF in the lung tissue of ARDS group was significantly reduced,and the positive expression of VCAM-1 and ET-1 was significantly increased.The expression of VEGF was significantly increased,and VCAM-1 and ET-1 positive expression was significantly reduced after amiloride intervention.(5)Western blot results showed that there was no significant change in PI3 K and Akt protein expression in the lung tissue of ARDS rats,and the expressions of uPAR,p-PI3 K and p-Akt and GSK 3? were significantly increased.After the intervention of amiloride,the expression of uPAR,p-PI3 K,p-Akt and GSK 3? were significantly reduced.Conclusion(1)Amiloride has protective effects on lung injury in ARDS rats;(2)The PI3 K / AKT / GSK 3? signaling pathway is involved in lung injury in ARDS rats,and amiloride can improves lung injury in ARDS rats by inhibiting PI3 K / AKT signaling pathway.