The Negative Regulation Of SOCS-3in JAK/STAT Signal Transduction Pathway Induced By TAP In Rat Pancreatic Acinar Cells

Objective: Through the establishment of rat pancreatic acinar cell inflammatory reactionmodel in vitro, while verifying the gene and protein expression changes of these relatedinflammatory factor, such as: JAK2, STAT3, and SOCS3, and the changes of TNF-αconcentration, so that we can confirm the negative regulatory role of SOCS-3inJAK/STAT signal transduction pathway. Above all, we can provide a theoretical basis forthe SOCS-3in the treatment study of acute pancreatitis.Methods:90SD rats after death, separation pancreatic pancreatic acinar cells, the cellsuspension was divided into5groups：(1) the control group, only add the same dose ofsaline;(2) experimental group1: add the TAP, pancreatic acinar cells were inducedinflammation;(3) the experimental group2: add the AG490pretreatment30min earlierbefore TAP;(4) the experimental group3: add the rapamycin pretreatment30min earlierbefore TAP;(5) experimental group4: add the SOCS-330min earlier before TAP. Everylarge group separately was divided into four subgroups according to the time point of TAPintervention1h,2h,3h,6h,1.5ml per subgroups. At above time points were sampled,real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) andprotein imprinting method (Western blot)1h,2h,3h point cells JAK2and STAT3,SOCS-3mRNA expression and protein expression; Enzyme coupling immune adsorbent(ELISA) determination of3h,6h time point in the upper liquid of TNF alpha factorconcentration. Statistical analysis used SPSS19.0software.Results:(1) Compared with control group, the mRNA relative expression and proteinexpression of JAK2,STAT3and SOCS3and the concentration of TNF alpha inexperimental group1was increased at each time point, there was statistically significant (P<0.01)，and under the stimulus TAP JAK2and STAT3, SOCS3mRNA1h activation (therelative expression increase),2h to the peak,3h activation showed a trend of decrease;JAK2, STAT3and SOCS3protein expression1h began to rise, and over time (2h,3h) ison the rise.(2) experimental group2, group3, group4compared with group1, the mRNA relative expression and protein expression of JAK2and STAT3, SOCS3was decreased ateach time point, with statistical significance (P <0.01); the concentration of TNF alphawas decreased at each time point, with statistical significance (P <0.05, P <0.01);(3) theexperimental group4compared with the experimental group2, group3, the mRNArelative expression of JAK2, STAT3was decreased at2h,3h time point, with statisticalsignificance (P <0.01); JAK2and STAT3protein expression was decreased at each timepoint, with statistical significance (P <0.01); TNF alpha concentration was decreased at6h time point, with statistical significance (P <0.01);(4) the experimental group2comparedwith the experimental group3, except SOCS3mRNA expression of relative rise at2h,3hpoint, with statistical significance (P <0.05, P <0.01), all of the other have no statisticalsignificance (P>0.05).Conclusion:①TAP induced the activation of pathway JAK2/STAT3signal in pancreaticacinar cell (i.e. the increase of JAK2, STAT3mRNA and protein expression), the factorinducing the release of TNF alpha and SOCS3;②SOCS3, AG490and rapamycin couldinhibit the activation JAK2/STAT3signaling pathway of pancreatic acinar cells which wasinduced by TAP, that is JAK2, STAT3mRNA and protein expression and TNF alpharelease;③Inhibitory effect of SOCS3are better than AG490. and rapamycin...