OB-RGRP regulates the phosphorylation of JAK2 and STAT3 in primary rat adipocytes

Abstract

This study aimed to explore the role of obestatin R gene-related protein (OB-RGRP) in autocrine signal transduction of adipocytes. Primary rat adipocytes were isolated and verified by microscopic observation and oil red O staining. OB-RGRP expression vector and OB-RGRP siRNA (si-OB-RGRP) were constructed and transfected into adipocytes. Adipocytes were then divided into five groups: (1) Control; (2) Vector (empty expression vector); (3) OB-RGRP (OB-RGRP expression vector); (4) si-OB-RGRP NC (si-OB-RGRP negative control); (5) si-OB-RGRP. mRNA and protein levels of OB-RGRP, JAK2, phosphorylated JAK2 (p-JAK2), STAT3 and phosphorylated STAT3 (p-STAT3) were examined using RT-PCR and western blot, respectively. Results showed that mRNA and protein levels of OB-RGRP in the Vector and si-OB-RGRP NC groups were similar to those in the Control group. Their levels in the si-OB-RGRP and OB-RGRP groups were significantly down-regulated and up-regulated (p < .05), respectively, in comparison with the control cells. There was no significant difference in the mRNA and protein levels of JAK2 and STAT3 among various groups. Moreover, the si-OB-RGRP NC and Vector groups induced similar ratios of p-JAK2 to JAK2 (p-JAK2/JAK2) and p-STAT3 to STAT3 (p-STAT3/STAT3) to the Control group. However, these two ratios in the si-OB-RGRP and OB-RGRP groups were significantly reduced and elevated (p < .05), respectively, in comparison with the Control group. These results suggested that OB-RGRP overexpression enhanced the levels of p-JAK2 and p-STAT3 while OB-RGRP silencing lowered their levels. In conclusion, OB-RGRP regulated the phosphorylation of JAK2 and STAT3 in primary rat adipocytes.