Transcriptional Research Of MRNA And LncRNAs In Copd Rat Model Of Lung Qi Deficiency Syndrome

Objective To explore potential biomarkers of chronic obstructive pulmonary disease(COPD)lung qi deficiency syndrome and provide theoretical and practical basis for the objective study of COPD lung qi deficiency syndrome,we performed RNA-seq technology and bioinformatics analysis to investigate the potential biomarkers between normal rats and COPD lung qi deficiency rats in the lay of messenger RNA(m RNA)and long noncoding RNA(Lnc RNA).Methods1.Mimic of COPD lung qi deficiency rat modelTwenty specific pathogen free(SPF)SD rats were randomly divided into control group and model group.We mimicked the COPD rat model of lung qi deficiency based on previous team experience in replicating models,which is via the combination of cigarette smoke(CS)and tracheal instillation of lipopolysaccharide(LPS).According to the pulmonary function,pathology of lung tissues,cough,wheezing,respiration,the glossiness of hair,etc.,to evaluate the establishment COPD rat model of lung qi deficiency.2.RNA sequencing and bioinformatic analysisThe Illumina Hi Seq 4000 sequencing platform was performed to detect the expression of m RNA and Lnc RNA.Based on the sequencing results,bioinformatic analysis methods like RNA coding ability prediction,RNA quantitative analysis,inter-group difference analysis,cluster analysis,Lnc RNA target gene prediction,and gene annotation were applied to obtain the differential expression of m RNA and Lnc RNA and predict the target genes of Lnc RNA and their potential functions.Results1.Compared with the rats of the control group,the rats in the COPD lung qi deficiency performed poor mental status,shortness of breath,cough,wheezing,hair dullness,etc.2.Pulmonary function results indicated that,compared with the control group,the pulmonary function parameters,forced vital capacity(FVC),forced expiratory volume in 0.3 seconds(FEV0.3),and FEV0.3/FVC,were significantly decreased(P<0.05),which were consistent with changes of COPD lung function.Also,the pathological results showed that the model group displayed expansion and fusion of alveolar,lung septal rupture,and emphysema.3.The expressions of serum IL-10,IL-23,and IL-32: the IL-10,compared with the control group,was down-regulated in the model group(P<0.05);however,the IL-23 and IL-32 were up-regulated in the model group(P<0.05).(1)We observed 550 known m RNA,237 novel m RNAs,753 known Lnc RNA,and134 novel Lnc RNA had observed significantly different expression in the comparison between the control group and the model group.(2)The target gene prediction analysis of significant difference expressed m RNA and Lnc RNA showed that a total of 109 pairs of Lnc RNA-m RNA interactions,involving72 lnc RNA and 90 m RNA.(3)GO enrichment analysis indicated that m RNA,which was significantly expressed between control group and model group,were enriched in(1)biological process which mainly contained cell processes(1016 m RNA),biological regulation(760 m RNA),metabolic processes(765 m RNA),biological process regulation(735 m RNA),etc.;(2)cellular component which mainly contained cells(1054 m RNA),cellular components(1053 m RNA),organelles(866 m RNA),etc.;(3)molecular function which mainly contained binding(941 m RNA),catalysts(412 m RNA),etc.(4)The KEGG enrichment analysis showed that significant differences of Lnc RNA targeting genes between groups were mainly distributed in the adipocytokine signaling pathway,protein digestion and absorption,fat digestion and absorption,NF-κB signal pathway,and inflammatory mediators in the TRP channel.(5)According to the Lnc RNA target gene prediction and enrichment analysis,LTCONS＿00047837,LTCONS＿00048719,LTCONS＿00048721,NONRATT017677.2,LTCONS＿00048720,LTCONS＿00048721,LTCONS＿00048723,NONRATT028939.2,LTCONS＿00062784,NONRATT028939.2 were involved in fat and protein related pathways via targeting m RNA to regulated the metabolism of nutrients.Conclusion1.The RNA-seq technology and bioinformatics analysis method had found that a large number of differentially expressed m RNA and Lnc RNA were observed in COPD lung qi deficiency syndrome,and they were predicted participating in COPD nutritional metabolism,immune inflammation,etc.2.Among the differentially expressed Lnc RNA,LTCONS＿00047837,LTCONS＿-00048719,LTCONS＿00048721,NONRATT017677.2,LTCONS＿00048720,LTCONS＿00048721,LTCONS＿00048723,NONRATT028939.2,LTCONS＿00062784,NONRATT028939.2 might be biomarkers of COPD lung qi deficiency,which a-ffected the immunity by regulating the body’s nutrient metabolism,and particip-ated in the development of COPD lung qi deficiency.