The Relationship Between Levels Of Plasma NOV/CCN3 With MCP-1 And IR Of NAFLD Patients

Background:Nonalcoholic fatty liver disease(NAFLD)is considered as one of the most common liver diseases.It is robustly linked to obesity and insulin resistance and is regarded as hepatic manifestation of metabolic syndrome(MetS).It has been shown that adipokines are involved in the pathophysiology of liver diseases.The nephroblastoma overexpressed gene(NOV)is a founder of the CCN(Cyr61/CCN1,CTGF/CCN2,NOV/CCN3)family of multitasking matricial proteins which regulate several cellular functions,such as proliferation,adhesion,differentiation,migration,survival,wound healing,angiogenesis.In addition,these proteins play key roles in several pathological conditions,such as inflammation,wound healing,fibrosis and cancers.We have recently shown that NOV is an new adipokine whose plasma level is positively correlated with BMI and fat mass.The Study on NOV-/-mice has shown that NOV can affect insulin sensitivity in a variety of ways.We know IR is an independent risk factor for NAFLD,so we now want to preliminary study the plasma levels of NOV in NAFLD patients and its relationship with IR.Objective : By measuring the level of NOV protein in serum and its relationship with the inflammatory factor MCP-1 and IR in patients with NAFLD investigate the role of NOV protein in the progression of NAFLD.Methods: In this study we collected 50 cases of patients with nonalcoholic fatty liver disease(31 males,19 females),mean age 40.34±14.12 years as the experimental group who were diagnosed through clinical,laboratory,ultrasound in our hospital outpatient department and physical examination.at the beginning of June 2016 to the end of October 2016.30 cases of healthy people(17 males,13 females),with mean age of 37.17±9.64 years,were randomly selected as the control group.The experimental group should meet the following conditions:(1)No history of drinking or alcohol consumption <140g / week(F <70g / week).(2)Abdominal ultrasonography is consistent with the imaging features of diffuse fatty liver.(3)All patients gave written informed consent.Inclusion criteria: no history of viral hepatitis;no abnormal liver function.Exclusion criteria for all subjects: history of heavy drinking;viral hepatitis,drug-induced hepatitis,autoimmune hepatitis,Wilson’s disease,infection and other specific diseases that causes fatty liver disease;ahyperthyroidism or hypothyroidism,cancer,blood system disorders,obstructive sleep apnea,etc;polycystic ovary syndrome(PCOS)in pregnancy,lactation and oral contraceptives women.All the patients with NAFLD are divided into two groups according to the serum alanine aminotransferase(ALT)levels,namely simple fatty liver(NAFL)group of 17 cases and steatohepatitis(NASH)group of 33 cases.All the subjects’ gender,age,occupation,past history(heart,brain,liver,kidney and other diseases),family history,alcohol consumption,smoking history,medication history,eating habits,etc.were investigated and recorded.From the waist to height,weight were measured specially,then calculated the body mass index(Kg)BMI= weight / height(cm2),total physical examination,abdominal CT examination if necessary.The subjects were fasted for more than 12 hours,and their venous blood were collected on the morning.Fasting blood glucose(FBG),fasting insulin(FINS),serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST),triglyceride(TG),total cholesterol(CHOL),low density lipoprotein cholesterol(LDL-C),high density lipoprotein cholesterol(HDL-C)and other biochemical indicators were detected by our hospital automatic biochemical analyzer.Calculated according to the formula: HOMA-IR(insulin resistance index)=fasting glucose levels(FBG,mmol/L)×fasting insulin levels(FINS,m IU/L)/22.5.Another venous blood 5ml was collected.Plasma NOV and MCP-1 were detected by enzyme linked immunosorbent assay(ELISA).According to the results of the examination,the relationship between plasmaNOV level and each index was analyzed,and its significance in the pathogenesis of NAFLD was speculated.SPSS21.0 version of statistical software was used for data processing and statistical analysis.Measurement data in line with the normal distribution were presented as mean±standard deviation.Data with skewed distributions(NOV,MCP-1)were equally summarized as the median and Q1–Q3 intervals and log-transformed before further analyses.Comparisons between the means of the two groups were performed using an unpaired Student’s t-test.Pearson’s or spearman correlation coefficients were computed to assess the associations between continuous variables and plasma NOV levels.Logistic regression analysis was used in the degree of correlation analysis.P <0.05 was considered statistically significant.Results:1 The comparison of general clinical data in the two groups: there was no significant difference(P>0.05)in age and years between experimental group and control group.Compared with the control group,BMI(29.63±2.82kg/m2 vs.21.16±2.04kg/m2)was higher in experimental group,and the difference was statistically significant(P<0.01).2 Comparison of blood biochemical indexes in the two groups: compared with the control group,the CHOL(5.14±0.67mmol/L vs.3.90±0.70mmol/L),TG(2.03±0.71mmol/L vs.0.69±0.22mmol/L),LDL(3.34±0.55mmol/L vs.2.10±0.57mmol/L)were higher in the experimental group.However the HDL(1.08±0.17mmol/L vs.1.39±0.28mmol/L)was lower in the experimental group,and all the differences were statistically significant(P<0.01).3 Comparison of HOMA-IR levels in the two groups: compared with the control group,the HOMA-IR was significantly higher(3.90±1.43 vs.1.98±0.98)in the experimental group,and the difference was statistically significant(P< 0.01).4 Comparison of plasma NOV and MCP-1 levels in two groups: the plasma levels of NOV(16.66±6.13ng/ml vs.8.00±2.08ng/ml)and MCP-1(62.79±30.58pg/ml vs.47.18±15.98pg/ml)in the NAFLD group weresignificantly higher than those in the control group,and the difference was statistically significant(P< 0.01).5 Correlation analysis of plasma NOV with MCP-1,HOMA-IR,BMI and biochemical indexes: NOV was positively correlated with MCP-1,HOMA-IR and BMI in experimental group(r value was 0.554,0.644,0.445,P was<0.01),MCP-1 and HOMA-IR was also positively correlated(r value was 0.466,P<0.01),however there was no significant difference between NOV with CHOL,TG,HDL,LDL(P>0.05).6 Comparison of serum NOV,MCP-1,HOMA-IR and lipids levels between NAFL group and NASH group: there were no significant difference of NOV levels(16.06±5.49ng/ml vs.17.82±7.26 ng/ml),MCP-1 levels(60.53±31.24pg/ml vs.67.17±29.70pg/ml),HOMA-IR(3.24±1.36 vs.4.19±1.43)and CHOL,TG,HDL,LDL between the two groups(P>0.05).7 With the occurrence of non-alcoholic fatty liver considered to be the dependent variable,and the gender,age,HOMA-IR,TG,CHOL,LDL,HDL,MCP-1 and NOV as independent variables,Logistic regression analysis showed that NOV was the risk factor of nonalcoholic fatty liver disease(OR=3.40 P<0.05).Conclusion:1 Insulin resistance was present in NAFLD patients;2 The serum levels of NOV/CCN3 and MCP-1 was increased in NAFLD patients,and NOV/CCN3 was positively related to MCP-1,HOMA-IR,BMI;3 NOV may promote the occurrence of inflammation,but the effect on NASH is uncertain;4 Elevated plasma NOV is a risk factor for NAFLD.