Study On Long-chain Non-coding RNA And Risk Factors Related To Impaired Glucose Regulation

In the past 37 years,several nationwide epidemiological surveys have found that the prevalence of type 2 diabetes and impaired glucose regulation in China has been increasing year by year,while the awareness rate of diabetes is low,the rate of compliance is low,and there are many complications.It is bringing huge economic losses and health burdens to the country,society and even individuals.Although the results of clinical intervention trials showed that lifestyle adjustment or drug intervention can delay the progression of pre-diabetic population to clinical diabetes,it cannot completely block the deterioration of glucose metabolism.Once the intervention is removed,most patients with prediabetes will still progress to clinical diabetes.The prevention of type 2 diabetes should be preceded by the prevention of impaired glucose regulation(IGR),that is,zero grade prevention.Not only do we need to take effective measures to inhibit the transition from IGR to clinical diabetes,but more urgently,we need to take measures to prevent impaired glucose regulation in the IGR high-risk population,reduce the increase of prediabetes,and reduce the reserve of diabetes.There are many links involved in glucose metabolism,and the possible mechanisms leading to impaired sugar regulation are very complicated.We started with molecular biology of energy metabolism,epidemiological investigation and literature meta-analysis,to look for long-chain non-coding RNAs(LncRNAs)related to impaired glucose regulation,high-risk groups,and early intervention methods.Part 1 Identification of related long non-coding RNAs and mRNAs in subclinical hypothyroidism complicated with type 2 diabetes——Molecular Biology Research on the Function of Glucose Regulation Background and ObjectiveThyroxine and insulin are both hormones that regulate the body’s energy metabolism.Under the synergistic effect of epinephrine,thyroxine increases the local cAMP products of skeletal muscle,then activates glycolytic enzymes,muscle cell membrane sodium and potassium pumps,calcium ion channels and sarcoplasmic reticulum phosphoprotein activity,and enhances glucose uptake and utilization.Subclinical hypothyroidism is often accompanied by type 2 diabetes(T2D),but the pathological mechanism of subclinical hypothyroidism and subclinical hypothyroidism with T2D is still uncertain.LncRNA plays a key role in regulating mRNA splicing,transcription,translation,import,export and stability.We aimed to find potentially relevant long-chain non-coding RNAs(lncRNAs)and mRNA in these diseases.MethodTranscriptome sequencing was performed in three patients with subclinical hypothyroidism(S),three patients with subclinical hypothyroidism complicated with type 2 diabetes(SD),and three healthy controls(N).Differentially expressed mRNAs(DEmRNAs)and differentially expressed lncRNAs(DElncRNAs)were screened in S vs.N,SD vs.N,and SD vs.S group,and the nearby and co-expressed DEmRNAs of DElncRNAs were screened in S vs.N and SD vs.N.Moreover,Gene ontology(GO)and Kyoto Encyclopaedia of Genes and Genomes(KEGG)molecular pathway enrichment analysis of DEmRNAs and mRNA-lncRNA co-expressed pairs in three groups(S vs.N,SD vs.N and SD vs.S)were performed by Metascape.ResultIn total,465,1058,and 943 DEmRNAs were obtained in S vs.N,SD vs.N,SD vs.S,respectively,and 191 overlapping genes were obtained in S vs.N and SD vs.N group.According to GO and KEGG analysis,it is speculated that LAIR2,PNMA6A,SFRP2 are related to subclinical hypothyroidism,while GPR162,APOL4 and ANK1 are speculated to be related to subclinical hypothyroidism complicated with type 2 diabetes.A total of 50,100,and 88 DEIncRNAs were obtained in S vs.N,SD vs.N and SD vs.S,respectively.Two DElncRNA-nearby targeted DEmRNA pairs(involved two DElncRNAs and two DEmRNAs)and three DElncRNA-nearby targeted DEmRNA pairs(involved three DElncRNAs and three DEmRNAs)were obtained in the S vs.N and SD vs.N group,respectively。In addition,a total of 336 lncRNA-mRNA co-expression pairs were obtained in S vs.N group(|r|>0.95 and p<0.05).Meanwhile,2536 lncRNA-mRNA co-expression pairs were obtained in the SD vs.N group(|r|>0.95 and p<0.05).Several key DElncRNA-DEmRNAs were shared in both the S vs.N group and the SD vs.N group,including LOC105369772-KIR2DS4 and C4BPA,LOC105378122-KIR3DL2 and LOC100499194-CD79A.Combining with the interaction network of DElncRNA-DEmRNA,PAX8-AS1,co-expressed with KIR3DL1,was identified to function in subclinical hypothyroidism,and JHDM1D-AS1,co-expressed with ANK1,was deduced to play a role in subclinical hypothyroidism complicated with type 2 diabetes.ConclusionIn all groups(S vs N,SD vs N and S vs SD),we identified key genes in the top 20 differentially expressed mRNA and lncRNA.Among genetic factors,several genes play a role in SCH,including LAIR2,PNMA6A,SFRP2 and lncRNA-PAX8-AS1.ANK1 and its co-expressed lncRNA-JHDM1D-AS1 may affect the glucose regulation function and participate in the occurrence of SCH and T2D.Our study preliminarily screened out the lncRNA and mRNA involved in the glucose regulation mechanism,which may provide new clues for further exploration of the glucose regulation mechanism,SCH and the pathogenesis of SCH complicated with T2D.Part 2 Investigation of risk factors related to impaired glucose regulation Background and Objective:Type 2 diabetes(T2D)increases the risk of microvascular and macrovascular complications,thus placing a heavy burden on patients and society.The prevalence of type 2 diabetes(T2D)and impaired glucose regulation(IGR,pre-diabetes)is increasing year by year,but the proportion of patients with diabetes and IGR who were aware of their diabetes condition is low,the treatment rate is low.Among patients treated,the proportion of good controll is low.As the T2D reserve army,IGR is even more serious.IGR has been identified as an independent risk factor for acute coronary events.Therefore,the zero-level prevention of T2D,including screening of IGR high-risk groups and IGR-related risk factors,and active intervention to prevent the conversion of IGR high-risk groups to IGR,will help slow down the rapid growth of the number of diabetic patients.Method774 permanent residents(residence time≥2 years)aged 15 or above in Shimuyuan Community of Jinan City were randomly selected and signed informed consent form which were investigated by trained investigators according to the questionnaire.They were divided into normal group(N group),impired glucose regulation group(IGR group)and diabetes group(DM group)according to their previous medical history.SPSS22.0 was used for statistical analysis of the survey data.The product of the risk factor(OR-1)and its prevalence×100(risk momentum,RM)is used to evaluate the influence of this factor on the occurrence of IGR among community residents.ResultsThe number of risk factors related to type 2 diabetes in each group except age was 2.85±1.493 for men in DM group,3.87±1.522 for women,3.71 ± 1.266 for men in IGR group,3.81 ±1.424 for women,and 3.81 ±1.424 for men in N group.1.62±1.371,female 1.56±1.316.There was a significant difference between the N group and the other two groups,but there was no significant difference between the IGR group and the DM group.After removing the age factor,other risk factors related to type 2 diabetes,in the N group,the top three incidences were overweight,sedentary lifestyle,hypertension history,as blood sugar increased,in the IGT group and DM In the group,the incidence and ranking of cardiovascular diseases such as cardiovascular diseases and metabolic disorders such as hyperlipidemia are increasing.In both male and female groups,age,hypertension,family history of diabetes(FHD),dyslipidemia,obesity,cardiovascular and cerebrovascular disease(CVD)were positively correlated with IGR.The sedentary lifestyle of men is negatively correlated with IGR,and overweight in women is positively correlated with IGR.The number of risk factors for type 2 diabetes(T2D)per person in the N group was positively correlated with age,regardless of gender.Glucose status worsens with age and the number of risk factors.FHD is the risk factor with the strongest risk momentum among men and women.ConclusionAge,hypertension,FHD,dyslipidemia,obesity and CVD are risk factors for IGR.FHD has the highest correlation with IGR and the highest risk momentum.IGR prevention should start with young people,including controlling weight,maintaining physical activity,and preventing high blood pressure and dyslipidemia,especially in those with FHD.Part 3 The effect of resistance training on serum insulin-like growth factor 1(IGF-1)---Mechanisms of methods for improving glucose regulationBackground and ObjectiveThe post-exercise hyperinsulinemic-positive glucose clamp test showed that more than 85%of the glucose intake is borne by the skeletal muscles.Postprandial hyperglycemia is related to muscle insulin resistance.Exercise can increase the signal transduction of insulin in skeletal muscle cells,enhance the translocation of GLUT4 to the cell membrane,improve insulin-stimulated glucose uptake,and improve glucose metabolism.Many T2D clinical intervention trials have adopted exercise as an important intervention method.Insulin-like growth factor-1(IGF-1)can exert an insulin-like effect by binding to the IGF-1 receptor,which can reduce tissue insulin resistance and improve glucose metabolism.IGF-1 is negatively correlated with age,and the decrease in serum IGF-1 will increase the risk of type 2 diabetes.There are conflicting data on the effect of resistance training on serum IGF-1 levels.In order to resolve this contradiction,we conducted a systematic review and meta-analysis to determine the impact of resistance training on serum IGF-1 levels.MethodThis systematic review and meta-analysis was performed and reported in accordance with the Preferred Reporting Items for Systematic Review and Meta-analysis(PRISMA)guidelines.PubMed,Scopus,Web of Science,and Embase databases were systematically searched from their inceptions until 10 December 2019 for randomized controlled trials(RCTs)comparing individuals who underwent resistance training and control participants.Only publications in English were included.To be included into the systematic review and meta-analysis,studies had to fulfill the following criteria:1.Randomized control trials(RCTs)by design;2.Examining the intervention of resistance training versus control in more than two weeks of follow up;3.Reported mean and SD or SE of serum IGF-1 before and after trials;4.Adult participants(≥18 years old).Case series,observational studies,case reports and studies published as abstracts were excluded.Trials examining the combination of resistance training with other confounders(thus the single effect of resistance training could not be investigated)were also omitted.The references of all eligible publications and relevant reviews were handsearched for additional papers.If several articles published overlapping data,we included the most recent publication or the one with a longer follow-up period.Two investigators independently checked the title and abstracts.Any disagreement was resolved via discussion with a third investigator.In the next step,the authors independently screened the full texts and any discrepancy was resolved by a head reviewer.Finally,studies were included based on agreement amongst all three investigators.A standardized data extraction form was used for extracting information from all eligible trials.Subsequently,we applied the Cochrane Risk of Bias Assessment tool to evaluate the risk of bias for all included papers.Statistical analysis was executed using the Stata software.We applied a random-effects model and inverse variance to calculate the weighted mean difference(WMD)with 95%confidence intervals(CI).We used Cochran’s Q and I2 tests to assess heterogeneity in the results of the trials.Funnel plot asymmetry was used to discover any publication bias in the meta-analysis;and Egger’s and Begg’s tests regression test was used to assess funnel plot asymmetry.The possible impact of participants’ age and follow up length were measured by means fractional polynomial modelling(non-linear dose-response analysis)and meta-regression(linear dose-response analysis).A sensitivity analysis was applied to examine the impact of individual trials,in which the effect size was derived by removing one study at a time to observe the extent to which inferences could be influenced by a specific trial.ResultsWe first retrieved 764 records from the four databases,and removed 437 duplicated records.After screening by the titles and abstracts,269 records were excluded.The full texts of the remaining 58 articles were appraised for inclusion,whereby 22 RCTs with 33 arms met the inclusion criteria for meta-analysis.33 trials reported IGF-1 level as an outcome measure.The pooled estimate demonstrated a significant increase in IGF-1(WMD:10.34 ng/ml,95%CI:4.93,15.74,p=0.000,I2=90.3%)after resistance training compared with the control group.Subgroup analysis demonstrated that the increase in IGF-1 levels following resistance training was only statistically significant in treatment duration≤16 weeks(WMD:8.04 ng/ml),participants aged more than 60 years old(WMD:9.84 ng/ml);and in women(WMD:17.27 ng/ml).Subsequent analysis of the relationship between participants’age with plasma IGF-1 alterations revealed a U shape correlation in non-liner dose response,in which resistance training resulted in a declined IGF-1 level up to 40 years of age.Beyond 40 years old,the IGF-1 level was increased following resistance training.ConclusionResistance exercise will affect serum IGF-1 level.People who receive training for≤16 weeks,participants over 60 years of age,and resistance training among women are associated with increased IGF-1 levels.There was a U-shaped association between participant age and changes in serum IGF-1.