Studies On Galectin-3 Correlates With Prognosis Of Hepatic Failure

Objective:By detecting levels of serum Galectin-3 in patients with hepatic failure(HF), to evaluate whether serum levels of Galectin-3 associated with the severity and prognosis of hepatic disease, and the effects of Galectin-3 on the prognosis of patients with HF. Meanwhile, to investigate the role of Galectin-3 combined with other HF prognostic indicators on the prognosis of HF.Methods:The patients with HF (HF group, n=76), patients with high jaundice (high jaundice group, n=38), patients with mild and moderate chronic hepatitis B (chronic hepatitis B group, n=26) were collected from inpatients in the department of liver disease the second hospital of Nanjing between January 2014 and December 2014, and normal control (control group, n=25) was collected from healthy people in the hospital during the same period of time. The serum levels of Galectin-3 were measured on admission, and were compared between each group. HF group was divided into survival group and death group according to the prognosis of the patients with HF at the end of follow-up. Observe the changes of serum Galectin-3 level before and after treatment between HF survival group and death group. Meanwhile, the general information, biochemical indices, complications and serum levels of Galectin-3 were compared by t test or Mann-Whitney U test between patients with HF survival group and death group, meaningful variables were analyzed by multiple factor Logistic regression. Finally, HF prognosis model was set up by using meaningful variables that were obtained from multiple Logistic regression analysis, and the optimal threshold of the model was determined.Results:The serum levels of Galectin-3 were detected by ELISA method, it showed that HF group, high jaundice group, chronic hepatitis B group, control group were 11.093±5.245 ng/ml,11.004±6.634 ng/ml,3.626±0.806 ng/ml,3.582±0.729 ng/ml. The serum levels of Galectin-3 in HF group was obviously higher than that of mild and moderate chronic hepatitis B group and control group, the difference was statistically significant(P<0.05). The difference of serum levels of Galectin-3 between HF group and high jaundice group had no statistical significance (P> 0.05).Before treatment, the difference of TBIL(P=0.028), ALB(P=0.018), INR(P=0.011), Na+(P=0-001), Bun(P=0.032), Cr(P=0.026), AFP(P=0.027), TG(P=0.033), TSH(P=0.001), HE(P=0.008), infection(P=0.001), ascites(P=0.000) and serum Galectin-3 (P=0.01) between HF survival group and death group had statistical significance. TBIL(P=0.011, β=-2.084), INR(P=0.036, β=-3.520), TSH(P=0.011, β=1.633), Galectin-3(P=0.011, β=0.43), ascites(P=0.001, β=-7.223) were related to the prognosis of HF by multiple Logistic regression analysis. Logistic regression model of HF prognosis:P= 1/(1+e-y), y=0.386-2.084X1-3.52X3+ 1.633X9+3.323X10-7.223X13 (P is the survival probability of patients with HF, X1, X3, X9, X10, X13 respectively represent TBIL, INR, TSH, Galectin-3 and ascites). The area under ROC curve of the regression model was 0.724, the prediction accuracy of the model was 85%.In dynamic observation, serum Galectin-3 level after treatment was obviously higher than before treatment in HF survival group(Z=-6.792, P=0.000), while serum Galectin-3 level between before treatment and after treatment had no obvious changes in HF death group, the difference was not statistically significant(Z=-0.665, P=0.506).Conclusions:Serum Galectin-3 was a protection factor for the prognosis of HF patients, it can be regarded as an indicator, which evaluates the prognosis of HF. Combining serum Galectin-3 with other clinical indicators can make a reasonable judgment on prognosis of HF.Objective:To build the specific Galectin-3 gene interference and over-expression lentivirus vectors and transfer normal human hepatocyte (HH). Observe the expression levels of Galectin-3 gene after interference and over-expression lentivirus vectors transferring HH, and observe the proliferative activity of HH and the expression levels of Wnt/β-catenin signaling pathway related proteins.Methods:This experiment was divided into five groups, interference group was Galectin-3 shRNA interference lentivirus transferred HH, and set the interference of negative control group. Over-expression group was Galectin-3 over-expression lentivirus transferred HH, and set the over-expression of negative control group. Blank group was HH in logarithmic phase without conducting. Observe the efficiency of infection under fluorescent inverted microscope after lentivirus transferring HH. the levels of Galectin-3 in five groups were detected by Western-blotting. The proliferative activity of HH was detected by CCK-8, and the levels of Axin, β-catenin, p-β-catenin, GSK-3β, p-GSK-3β were determined by Western-blotting.Results:After interference and over-expression lentivirus vectors transferred HH, the expression levels of Galectin-3 in interference group was more obviously declined than blank group and interference of negative control group(P< 0.05). While, the expression levels of Galectin-3 in over-expression group had a more significant increase than blank group and over-expression of negative control group(P< 0.05).After interference and over-expression lentivirus vectors transferred HH for 24h, 48h and 72h, the value of OD was detected by CCK-8 kits. The results showed OD of interference group was lower than blank group and interference of negative control group, the difference was not statistically significant (P< 0.05). There was no statistical significance between blank group and interference of negative control group(P> 0.05). OD of over-expression group was higher than blank group and over-expression of negative control group, the difference was not statistically significant (P< 0.05). There was no statistical significance between blank group and over-expression of negative control group(P> 0.05).The protein electrophoresis banding of p-GSK-3p, GSK-3β, p-β-catenin, β-catenin, Axin and β-actin were analyzed by Quantity One software. It showed that there were no significant differences in the protein expression levels of Axin and total GSK-3β, p-β-catenin between interference group and blank group, interference of negative control group(P> 0.05). The protein expression levels of p-GSK-3p., β-catenin in interference group were more obviously decreased than blank group and interference of negative control group(P< 0.001). There were no significant differences in the protein expression levels of Axin and GSK-3β, p-β-catenin between over-expression group and blank group, over-expression of negative control group(.P > 0.05). The protein expression levels of p-GSK-3β、β-catenin in over-expression group were obviously higher than blank group and over-expression of negative control group(P< 0.001).Conclusions:Galectin-3 can promote the regeneration of HH and its mechanism may be through up-regulation the protein expression of p-GSK-3β, β-catenin, Wnt/β-catenin signaling pathway.