In Vitro Synthesis Of JSRV-env MRNA And FGF21 MRNA With Explore Their Biological Function

Objective:JSRV-env and FGF21 m RNA were synthesized in vitro by in vitro transcription.To dectcted the specific antibodies produced by JSRV-env m RNA in mice,which reveale the therapeutic potential of m RNA in infectious diseases and explore the therapeutic effect of lung adenocarcinoma,and to lay the experimental foundation for further prevention of respiratory diseases in China.To detect the effect of FGF21 m RNA on glucose metabolism in hepatocytes of insulin resistance model,and to lay the foundation for further study on the mechanism of FGF21 in improving insulin resistance to hypoglycemia,Then to explore a new method for the treatment of T2 DM.Methods: To optimize the PT7 TS vector sequence,and inserted the target sequence then in vitro synthesized JSRV-env m RNA.Combined the m RNA with protamine subcutaneous injected into BALB/c mice;Using lentivirus packaging system to produce JSRV pseudovirus,The immunized mice were subjected to pseudovirus-based antibody neutralization assay to detect specific antibodies against JSRV-env m RNA in mice.FGF21 m RNA was synthesized in vitro.Hepatocytes were cultured in RPMI-1640 medium supplemented with 34.4 ?mol/L recombinant insulin and 1 ?mol/L dexamethasone in 10% FBS RPMI-1640 for 72 h to induce insulin resistance(IR)).IR cells were divided into IR model control group,IR insulin group,IR FGF21 group,IR insulin + FGF21 group.The glucose uptake was measured by glucose oxidase(GOD-POD)method.Real-time PCR was used to detect the expression of GLUT1 m RNA.The protein expression of GLUT1 in model cells was detected by Western blot.Results: JSRV-env m RNA and FGF21 m RNA were successfully synthesized in vitro.JSRV pseudovirus was reorganized.Pseudo-virus in vitro assay showed that JSRV-env m RNA-immunized mice produced specific antibodies against JSRV.After IR cells transfected of FGF21 m RNA.Compared with IR control group,glucose uptake of IR FGF21 group was increased(p<0.05),the expression of GLUT1 m RNA was significantly increased(p<0.05)and the protein expression of GLUT1 was significantly increased(p<0.05).Conclusion: JSRV-env m RNA and FGF21 m RNA were successfully synthesized in vitro.The JSRV-env m RNA can express to produce neutralizing antibodies against JSRV-env.FGF21 m RNA can improve the uptake of glucose in insulin resistance model cells.