The Study On Regulation Of PI3K/AKT/NF-KB Signaling Pathway On HIF-1α Expression Changes In Bone Marrow Tissues Of Rats With High Altitude Erythrocythemia

Objective:By replicating the pathological model of HAPC in rat plateau to verify the correlation between PI3K/AKT/NF-KB signaling pathway and HIF-1α expression change in myeloid tissue of HAPC rats,which provide experimental theoretical basis for systematically clarified the pathogenesis and treatment study of HAPC.Method : In this study,20 healthy male SPF-grade SD rats were selected and the weight was 250±10g,which divided into blank group and HAPC group(model group)randomly,each group of 10.DS-F type high quality disease environmental simulation system was used to simulate the plateau environment with altitude of 3500m~6000m,and the model time was 60 d.The whole blood was used for routine blood test after the model was finished;the content of sterum EPO was detected by ELISA assay;bone marrow histopathology observed under microscope;the protein of P-AKT、NF-kB P65 and HIF-1αin rat bone marrow tissue were detected by Western Blot and IHC assay;the levels of P-AKT、NF-kB P65、HIF-1α mRNA were detected by RT-PCR assay.Result:1.the blood routine test results showed that the HGB content range of the HAPC group was between 235 and 273 g/L,which was higher than 210g/L,indicating the success of the model.2.Compared with the blank group,the serum EPO content in the HAPC group was significantly increased(P < 0.01),suggesting that the model was successful.3.the histopathology observation of rat bone marrow showed that compared with the blank group,HAPC group of trabecular bone structure is normal,but the bone marrow cavity capillary number and cell number increased obviously,capillary internal bleeding,part of the expansion,megakaryocyte increased slightly,observed under 100 times microscope;the bone marrow cavity cell arranged densely,capillary seedless red blood cells increased obviously,the erythrocytes increased obviously,the proportion of granule cells decreased and a small number of granulocytes enter the blood sinus,observed under 400 times microscope.4.The WB results all showed that the proteins of P-AKT,NF-kB p65 and HIF-1α were significantly increased in the bone marrow tissues of the HAPC group(P < 0.01),compared with the blank group.5.The IHC results showed thatthe proteins of P-AKT,NF-kB p65 and HIF-1α were significantly increased in the bone marrow tissues of the HAPC group(P < 0.01),compared with the blank group.6.The RT-PCR results showed that the genes of P-AKT,NF-kB p65 and HIF-1αmRNA in the bone marrow tissues of the HAPC group were significantly increased(P < 0.05).Conclusion: 1.The pathological feature of myeloid tissue in HAPC rats showed that the total number of cells,erythroid cells,blood sinus dilation,congestion and blood sinus quantity in marrow cavity increased significantly.2.the protein and mRNA levels of P-AKT 、 NF-kB p65 、 HIF-1α in HAPC rats were significantly increased.3.The PI3K/AKT/NF-KB signaling pathway was positive regulatory with HIF-1α expression change in myeloid tissue of HAPC rats.