Background

Protein kinase D (PKD) family, which includes PKD/PKD1, PKD2, and PKD3, has been increasingly implicated in the regulation of multiple cellular functions and human diseases. We recently reported that pharmacologic inhibition of PKD ameliorated the pathologic responses and severity of pancreatitis. However, to further investigate the importance of PKD family members in pancreatitis, it is necessary to explore the effects of pancreas-specific genetic inhibition of PKD isoform on pathology of pancreatitis.

Methods

We generated a mouse model (referred as PKD3Δpanc mice) with pancreas-specific deletion of PKD3, the predominant PKD isoform in mouse pancreatic acinar cells, by crossing Pkd3flox/flox mice with Pdx1-Cre transgenic mice which express Cre recombinase under the control of the mouse Pdx1 promoter. Pancreas-specific deletion of the PKD3 gene and PKD3 protein was confirmed by PCR and Western blot analysis. Experimental pancreatitis was induced in PKD3Δpanc and Pkd3flox/flox (control mice) littermates by intraperitoneal injections of cerulein or L-arginine.

Results

Compared to the control mice, PKD3Δpanc mice displayed significant attenuation in inflammation, necrosis, and severity of pancreatitis in both experimental models. PKD3Δpanc mice had markedly decreased NF-κB and trypsinogen activation, pancreatic mRNA expression of multiple inflammatory molecules, and the receptor-interacting protein kinase 1 (RIP1) activation in pancreatitis. PKD3Δpanc mice also had less pancreatic ATP depletion, increased pro-survival Bcl-2 family protein expression, and autophagy promotion.

Conclusion

With PKD3Δpanc mouse model, we further demonstrated that PKD plays a critical role in pathobiological process of pancreatitis and PKD constitutes a novel therapeutic target to treat this disorder.

中文翻译：

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蛋白激酶 D 的胰腺特异性缺失减轻急性胰腺炎的炎症、坏死和严重程度

背景

蛋白激酶 D (PKD) 家族，包括 PKD/PKD1、PKD2 和 PKD3，越来越多地参与调节多种细胞功能和人类疾病。我们最近报道了 PKD 的药理学抑制改善了胰腺炎的病理反应和严重程度。然而，为了进一步研究 PKD 家族成员在胰腺炎中的重要性，有必要探讨胰腺特异性基因抑制 PKD 亚型对胰腺炎病理学的影响。

方法

我们通过将Pkd3flox/flox小鼠与表达 Cre 重组酶的 Pdx1-Cre转基因小鼠杂交，生成了一个小鼠模型（称为 PKD3 Δpanc小鼠），其中 PKD3 是小鼠胰腺腺泡细胞中主要的 PKD 异构体的胰腺特异性缺失。小鼠Pdx1启动子。PKD3 基因和 PKD3 蛋白的胰腺特异性缺失通过 PCR 和蛋白质印迹分析得到证实。通过腹膜内注射雨蛙素或 L-精氨酸在 PKD3 Δpanc和Pkd3flox/flox（对照小鼠）同窝小鼠中诱导实验性胰腺炎。

结果

与对照小鼠相比，PKD3 Δpan c 小鼠在两种实验模型中都显示出炎症、坏死和胰腺炎严重程度的显着减弱。PKD3 Δpan c 小鼠在胰腺炎中显着降低了 NF-κB 和胰蛋白酶原的激活、多种炎症分子的胰腺 mRNA 表达以及受体相互作用蛋白激酶 1 (RIP1) 的激活。PKD3 Δpan c 小鼠的胰腺 ATP 消耗较少，促存活 Bcl-2 家族蛋白表达增加，自噬促进。

结论

通过 PKD3 Δpanc小鼠模型，我们进一步证明了 PKD 在胰腺炎的病理生物学过程中起着关键作用，并且 PKD 构成了治疗这种疾病的新治疗靶点。