GSK-3β/β-catenin信号通路在氯氮平导致的胰腺损伤中的作用及其机制研究

Schizophrenia (SZ) is a serious mental disease and harmful to human health, one third of which is refractory. Clozapine is the best treatment for SZ, but its side effects are also very serious. Pancreatitis is one of the fatal side effects induced by clozapine. The acute pancreatitis is a disease defined as acute inflammatory process and necrosis of the pancreas characterized by premature activation of digestive enzymes within the pancreatic acinar cells and causing pancreatic auto-digestion. The typical pancreatitis is well-known. But asymptomatic pancreatitis has rarely been reported up to now and is not recognized as a typical side-effect of clozapine. According to one of the FDA reports of pancreatitis occurring in conjunction with antipsychotic agents, 40% patients were receiving treatment with clozapine. Clozapine can cause pancreatitis and pancreatic beta-cell apoptosis, suppression of cell proliferation, and trends of reduction in pancreatic insulin content, however the mechanism of clozapine on pancreatic damage is not clear. Our previous studies showed that clozapine can regulate inflammatory cytokines, and inflammatory cytokines play an important role in pancreatitis. In the study of pancreatic damage animal model,our results indicated macrophages infiltration and the increase of cytokines and chemokines.In addition, it showed protein oxidation, lipid peroxidation and the expression of iNOS, indicating oxidative stress.Further studies showed that clozapine can affect GSK-3β. Our research hypothesis is that the GSK-3β/β-catenin signaling pathway plays an important role in the regulation of pancreatic damage induced by clozapine. Therefore, in order to clarify the relationship between clozapine and pancreatic damage, inflammatory cytokines and signal pathway, we use the clinical research and establish pancreatic damage animall model induced by clozapine and detect the pancreatic damage index (blood amylase, pancreatic cell apoptosis and necrosis, etc.), Inflammatory cytokines (TNF-α、IL-1β and IL-6) and signal pathway genes (GSK-3β、β-catenin、 NF-κB, etc.).At the same time , the pancreatic damage index and GSK-3β/β-catenin signaling pathway will be detect to clarify the role and mechanism of GSK-3β/β-catenin Signaling pathway in the pancreatic damage induced by clozapine.

精神分裂症（SZ）是一种严重危害人类健康的精神疾病，其中难治性SZ占三分之一。氯氮平是治疗SZ最好的药物，但其副作用也很严重。氯氮平导致胰腺损伤的作用机制一直不清楚。我们之前的研究结果显示，氯氮平可以影响炎性因子，炎性因子在胰腺炎中起到重要作用，进一步研究发现氯氮平可以影响GSK-3β。本研究假设GSK-3β/β-catenin信号通路在氯氮平导致的胰腺损伤中具有重要调控作用。因此，本研究通过临床研究和氯氮平胰腺损伤动物模型研究，检测胰腺损伤指标(血淀粉酶、胰腺细胞凋亡坏死等)、炎症因子（TNF-α、IL-1β和IL-6等）和信号通路基因（GSK3β、β-catenin、 NF-κB等）。明确氯氮平与胰腺损伤、炎症因子和信号通路的关系，阐明GSK-3β/β-catenin信号通路在氯氮平对胰腺损伤中的调控作用。

氯氮平是治疗精神分裂症疗效最好的药物，但其严重的副作用制约了临床应用，因此探讨氯氮平副作用产生的机制，对于临床合理使用氯氮平意义重大。在此背景下我们通过临床和基础研究来探讨氯氮平对胰腺损伤的副作用机制。首先，通过临床横断面的对照研究发现，服用氯氮平的患者与正常健康对照组相比，外周血炎症因子IL-6水平和外周血单核细胞GSK-3β、β-catenin信号蛋白表达均升高。其次，通过小鼠高中低三种剂量氯氮平动物模型研究发现（1）胰腺外分泌腺的指标血淀粉酶的水平随着氯氮平的给药剂量的增加而明显上升，而内分泌腺各项指标并不一致，相互关系有待进一步探究。（2）检测反应胰腺损伤的指标（Caspase3、ATF6、CHOP、CD68、和HMGB1），均随着氯氮平的给药剂量的增加而明显上升。（3）GSK-3β与β-catenin的表达与炎症因子（TNF-α、IL-1β和IL-6）水平随着胰腺损伤加重而升高。综上，本课题研究发现氯氮平会导致胰腺功能损伤，炎症因子水平改变，以及GSK-3β/β-catenin通路表达升高。三者之间具有相关性，且损伤具有剂量相关性。说明氯氮平的治疗作用以及副作用与GSK-3β/β-catenin通路密切相关。该研究为进一步了解氯氮平作用机理，如何在临床避免其不良反应而充分发挥其治疗作用奠定一定的工作基础。对于今后否可以通过调节 GSK-3β的表达来减轻氯氮平导致的胰腺损伤的严重程度，有待进一步研究。本项目资助发表论文21篇（含会议论文12篇），待发表5篇。培养硕士生7名，其中3名已经取得硕士学位，4名在读。项目投入经费25万元，支出20.52万元，各项支出基本与预算相符。剩余经费4.48万元，剩余经费计划用于本项目研究后续支出。