Involvement of M3 cholinergic receptor signal transduction pathway in regulation of the expression of chemokine MOB-1, MCP-1 genes in pancreatic acinar cells

Abstract

Whether M3 cholinergic receptor signal transduction pathway is involved in regulation of the activation of NF-κB and the expression of chemokine MOB-1, MCP-1 genes in pancreatic acinar cells was investigated. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, atropine and PDTCin vitro. The MOB-1 and MCP-1 mRNA expression was detected by using RT-PCR. The activation of NF-κB was monitored by using electrophoretic mobility shift assay. The results showed that as compared with control group, M3 cholinergic receptor agonist (10⁻³ mol/L, 10⁻⁴ mol/L carbachol) could induced a concentration-dependent and time-dependent increase in the expression of MOB-1, MCP-1 mRNA in pancreatic acinar cells. After treatment with 10⁻³ mol/L carbachol for 2 h, the expression of MOB-1, MCP-1 mRNA was strongest. The activity of NF-κB in pancreatic acinar cells was significantly increased (P<0.01) after treated with M3 cholinergic receptor agonist (10⁻³ mol/L carbachol)in vitro for 30 min. Either M3 cholinergic receptor antagonist (10⁻⁵) mol/L atropine) or NF-κB inhibitor 10⁻² mol/L PDTC) could obviously inhibit the activation of NF-κB and the chemokine MOB-1, MCP-1 mRNA expression induced by carbachol (P<0.05). This inhibitory effect was significantly increased by atropine plus PDTC (P<0.01). The results of these studies indicated that M3 cholinergic receptor signal transduction pathway was likely involved in regulation of the expression of chemokine MOB-1 and MCP-1 genes in pancreatic acinar cellsin vitro through the activation of NF-κB.

Keywords

pancreatic acinar cell / M3 cholinergic receptor signal transduction pathway / chemokine / NF-κB

References

[1] HanB, JiB, LogsdonC D, et al. . CCK independently activates intracellular trypsinogen and NF-kappaB in rat pancreatic acinar cells. Am J Cell Physiol, 2001, 280(3): C465-C465

[2] TandoY, AlotilH, SchneiderG, et al. . Induction of Iκ B-kinase by cholecystokinin is mediated by trypsinogen activation in rat panreatic lobules. Digestion, 2002, 66237-237

[3] CohenM C S C. Cytokine function: a study in biologic diversity. Am J Clin Pathol, 1996, 105598-598

[4] MayerJ, RanB, GansaugeF, et al. . Inflammatory mediators is human acute pancreatitis: clinical and pathophysiological implications. Gut, 2000, 47546-546

[5] BradyM, BhatiaM, ChristmasS, et al. . Expression of the chemokines MCP-1/JE and cytokine-induced neutrophil chemoattractant in early acute pancreatitis. Pancreas, 2002, 25(3): 260-260

[6] TerrenceG, PengL, StephenA, et al. . Chemokine gene expression in rat pancreatic acinar cell is a early event associated with acute pancreatitis. Gastroenterology, 1997, 1131966-1966

[7] GronroosJ M, LaineJ, KailaT, et al. . Chronic alcohol intake and carbachol-induced acute pancreatitis in the rat. Exp Toxicol Pathol, 1994, 46(2): 163-163

[8] LogsdonC D. The influence of the cellular context on receptor function: a necessary consideration for physiologic interpretations of receptor expression studies. Life Sci, 1999, 64(6–7): 369-369

[9] BlinmanT A, GukovskyI, MouriaM, et al. . Activation of pancreatic acinar cells on isolation from tissue: cytokine upregulation via p38 MAP kinase. Am J Physiol Cell Physiol, 2000, 279(6): C1993-C1993

[10] SamuelI, ZaheerS, FisherR A, et al. . Cholinergic receptor induction and JNK activation in acute pancreatitis. Am J Surg, 2003, 186(5): 569-569

[11] StefanW, IrvinM, LauraH, et al. . Telenzepine-sensitive muscarinic receptors on rat pancreatic acinar cells. Am J Physiol (Gastrointest Liver Physiol), 1998, 274(37): G734-G734