The Study Of The MiRNA Regulated Immune System After Elderly Hip Fracture Based On Gene Chip Technology

Objective:To screen the specific regulated miRNA by using miRNA Microarray in the immune disorders rats,as the elderly hip fracture complications warning marker,and provide new ideas for the clinical treatment for hip fractures in the elderly.Methods:Seventy-five SD rats were used in this study,male,12 months old.Fifteen rats as normal control,the other 60 were established elderly hip fracture model.The IL-10 and TNF-a value were measured 24h,48h and 72h after fracture,and based on the IL-10 and TNF-a value of 72h after fracture,the rats were divided into disorder group and recovery group.The plasma miRNA microarray were performed in three rats from each group.miRNA Microarray were used to find differentially expressed genes,and the results were verified by using Q-PCR for immune disorders organ disorder rats.And the specific miRNA regulation of immune disorders were selected.After that,through the microRNA targeting technique,to explore the relation of the microRNA and some signaling pathways,the key protein were also found and Western blot(WB)verification were performed.As the results showing,to explore the effects on the immune system and its effects on the corresponding organs.Results:Compared with the normal control group,the content of TNF-a in the hip fracture rats increased dramatically,showing acute inflammation,was the highest value at 24h,48h and declining after 72h;the content of IL-10 in the 24h showed a sharp increase in the trend,48h and 72h gradually restored.Between immune disorders group and the normal group 13 miRNA express the differences,six genes showed up-regulated and 7 genes were down.Real-time quantitative PCR method were used to verify the results,and the results showed that compared with normal control group,the plasma miR-130a-3p in immune disorder group were significantly lower(P<0.001),and the results showed similar trend in the lungs.By analyzing the miR-130a-3p regulation of immune system-related functions in the target protein,the IRF1 and S1pr1 protein were selected.In our results,the IRF1 expression in immune disorder group was significantly higher than which in the normal group,but no significant difference when compared with the recovery group and the normal group.As concern as the Slprl expression level,which were significantly higher in the immune disorders group and recovery group than which in normal group.Our analysis found that miR-130a-3p acts on the target sites of 3'UTR region 411-418 of IRF1,and 3'UTR 266-273 of Slprl.In the experiment,an immune disorder group showed the symptoms of become moribund,we examined the various organs of the pathological rats and found that the lungs of rats immune disorder group showed significant symptoms of inflammation,and this results also show that,the lungs miR-130a-3p regulation associated with inflammation.Conclusion:Though the detection of miRNA in elderly hip fractures rats,the results showed that the miR-130a-3p decreased significantly in the rats with immune disorders,and which increased the expression of TNF-?,S1pr1 and IRF1.Therefore,the use of miR-130a-3p as a potential biomarkers for clinical diagnosis and early warning is important.