Investigation of the involvement of immunity in the spontaneously occurring chronic pancreatitis in male Wisar Bonn/Kobori rats and human.

在本研究的第一部分，我们旨在探讨免疫机制，并探讨T细胞与腺泡细胞凋亡的关系，以及T细胞抑制剂他克莫司对雄性WBN/KOB大鼠慢性胰腺炎的预防作用。15周时小叶间结缔组织和实质内可见大量由F4/80阳性细胞(=单核/巨噬细胞)、CD4阳性细胞和CD8阳性细胞组成的细胞浸润物。尤其是CD8阳性细胞侵入胰腺小叶，并与腺泡细胞形成密切联系，部分腺泡细胞呈现凋亡特征。连续皮下注射他克莫司10周，可完全阻止20周龄胰腺腺泡细胞凋亡、CD_4、CD_8阳性细胞浸润及胰腺炎的发生。本研究旨在：1)定量测定雌二醇对腺泡细胞凋亡及慢性…的影响更多重症胰腺炎；2)评估其保护机制。20周龄WBN/KOB大鼠经雌二醇处理10周后，抗单链DNA抗体染色的腺泡细胞凋亡率、组织学评分和胰腺MPO活性均显著低于对照组。它还显著减轻了20周时胰腺羟脯氨酸含量的增加，这是胶原沉积的一个指标。雌二醇组大鼠胰腺组织中CD4、CD8T细胞数量明显减少。雌二醇可显著降低1%植物血凝素诱导的BrdU体外掺入脾细胞。20周龄WBN/KOB雄性Wistar大鼠经静脉注射WBN/KOB雄性大鼠脾细胞后，胰腺组织中雄激素受体未见表达，氟他胺饮食治疗对慢性胰腺炎无明显影响。总之，我们证明了T细胞，可能是CD8阳性细胞，参与了诱导腺泡细胞的凋亡，并提出了他克莫司可能应用于自身免疫性慢性胰腺炎的临床应用的可能性，以及雌二醇通过抑制T细胞的渗透和功能而剂量依赖性地抑制腺泡细胞的凋亡和慢性胰腺炎的发展。在目前的模型中，我们仍在通过评估新药对慢性胰腺炎和胰腺纤维化的影响来研究慢性胰腺炎的发病机制。较少

In the first part of the present study, we purposed to assess the immunologic mechanism and investigated T-cell association with acinar cell apoptosis and a preventive effect of tacrolimus, a T-cell suppressant, on the development o chronic pancreatitis in male WBN/Kob rats. At 15 week, cellular infiltrates composed of F4/80 positive cells (=monocytes/macrophages), CD4 positive cells, and CD8 positive cells were extensive in the interlobular connective tissue and parenchyma. In particular, CD8 positive cells invaded pancreatic lobules and formed close associations with acinar cells, some of which demonstrated features of apoptosis. Repeated subcutaneous injection of tacrolimus for 10 weeks completely prevented the occurrence of acinar cell apoptosis, infiltration of CD4 and CD8 positive cells, and development of pancreatitis at the age of 20 weeks.The second study was conducted with the aim of : 1) quantitatively determining the influence of estradiol on acinar cell apoptosis and chron … More ic pancreatitis ; and 2) assessing the mechanism of its protection. Treatment with estradiol for 10 weeks significantly decreased the number of apoptotic acinar cells stained with an anti-single strand DNA antibody, histological scores, and pancreatic MPO activity at 20 week old WBN/Kob rats as compared with the control values. It also significantly attenuated the increase in pancreatic hydroxyproline content, an indicator of collagen deposition, at 20 weeks. Estradiol caused significant decrease in the numbers of CD4 and CD8 T cells in the pancreas. Estradiol significantly reduced 1 % phytohemagglutinin-induced incorporation of BrdU into the splenocytes in vitro. Androgen receptors could not be immunohistochemically identified in the pancreas at 20 weeks and dietary treatment with flutamide did not influence the chronic pancreatitis.We also tried to induce chronic pancreatitis in male Wistar rats by intravenously injecting splenocytes derived from 20 week old male WBN/Kob rats and to assess the gene expression by RT-PCR in the biopsy samples obtained from the human pancreas using endoscopic ultrasound technique. However, these trials has resulted in failure.In conclusion, we demonstrated that T cells, possibly CD8 positive cells, are involved in inducing apoptosis of acinar cells and raises the possibility that tacrolimus might find clinical application in the treatment of autoimmune chronic pancreatitis and that estradiol dose-dependently attenuates acinar cell apoptosis and development of chronic pancreatitis by suppressing infiltration and function of T cells. We are still conducting the investigations regarding the pathogenesis of chronic pancreatitis by assessing the effect of new drug on the chronic pancreatitis and pancreatic fibrosis in the present model. Less