Studies on mechanism of pancreas, especially β-cell, regeneration using surgical models

利用手术模型研究胰腺尤其是β细胞的再生机制

• 批准号: 12671158
• 负责人: SUMI Shoichiro
• 金额: $ 2.18万
• 依托单位: Shimane University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671158/
• 关键词: Pancreas regeneration; Pancreas duct ligature; pancreatectomy; Exocrine insufficiency; pancreatin; insulin; PDX-1; In situ hybridization

Pancreas remnant can regenerate after major pancreatectomy in rats. Gut hormones secreted in response to pancreatic exocrine insufficiency are considered involved deeply in the mechanism. On the other hand, pancreatic disorders induced by duct obstruction and pancreatitis induce expression of reg protein and various growth factors that activate regenerative process of the pancreas. However, little is known about the interactive effects of these intestinal and pancreatic factors on pancreas regenerationThis study investigated the role of these factors in rats using surgical models as follows. 65%Px or 65%Lig : Pancreas was tied off at ventral of the portal vein and the distal pancreas (splenic and gastric lobes) was resected (pancreatectomy abbreviated as Px) or left in the place (ligature abbreviated as Lig), respectively. 65%Sham : The pancreas was prepared but not tied. 80%Px or 80%Lig : In addition to 65%Px or Lig, duodenal lobe was tied off at a just anal level of common bite pancr … More eatic duct junction with duodenum and the distal pancreas (lower half of duodenal lobe) was resected (Px) or left in the place (Lig), respectively. 80%Sham : The pancreas was prepared but the ligatures were not placedEach group was fed either with normal powder diet or the diet supplemented with pancreatin (1/30 w/w), respectively. On postoperative day (POD) 8 and 21 (in 80% groups alone), rats were sacrificed and the pancreas was removed, weighed and histologically analyzed. The regeneration percent of the pancreas remnant was calculated on the basis of the mean value of body weight-normalized weight of the corresponding part in sham-operated groups. The percent of the pancreas remnant was 36.4 ± 1.4% in 65%Sham and 19.6 ± 0.7% in 80%Sham, respectively. Values are mean ± SE.Statistical significance was accepted when p<0.05On POD 8, wet weight of pancreas remnant was significantly increased in Px and Lig groups both in 65% and 80% experiments. Regeneration percent of Px/Lig/Sham in normal diet was 29 ± 7/14 ± 10/4 ± 6 in 65% and 24 ± 4/35 ± 13/5 ± 5% in 80%, respectively. Diet had no significant effect. On POD 21, pancreas weight was significantly increased only in Px group. Regeneration percent of Px/Lig/Sham in normal diet was 54 ± 8/11 ± 8/-13 ± 4%. Again, diet had no significant effect. The tissue was analyzed by insulin immuno-staining and in situ hybridization for PDX-1. No remarkable difference was noted in the pancreas remnant in both analysis. However, in the duct-ligated lobe of Lig group, diffuse expression of PDX-1 and a few insulin-positive cells were observed in the duct epithelium, suggesting activation of regeneration processIn conclusion, this study failed to show significant effect of pancreatic enzyme supplemented in the diet, suggesting that intestinal factor may not play a major role in pancreas regeneration. Clear difference in the results between POD 8 and POD 21 is very interesting. Further investigation of the cause of this difference seems to lead to profound understanding of the mechanism of pancreas regeneration Less

大鼠胰腺大部切除后残存胰腺可再生。胰腺外分泌功能不全时分泌的胃肠激素被认为与胰腺外分泌功能不全的机制密切相关。另一方面，胰管梗阻和胰腺炎引起的胰腺疾病会诱导REG蛋白和各种生长因子的表达，从而激活胰腺的再生过程。然而，这些肠道和胰腺因子对胰腺再生的交互作用知之甚少。本研究使用以下手术模型研究了这些因子在大鼠体内的作用。65%PX或65%LIG：分别在门静脉腹侧结扎胰腺，切除远端胰腺(脾、胃叶)或留在原位(LIG)。65%假：胰腺已做好准备，但未捆绑。80%PX或80%LIG：除了65%PX或LIG外，十二指肠叶仅在普通咬合PANCR…的肛门水平被捆绑较多的十二指肠胆管连接部和远端胰腺(十二指肠叶下半部分)分别被切除(Px)或留在原位(Lig)。80%假手术组：制备胰腺，但不放置结扎线。每组分别饲喂普通粉末饲料和添加胰酶的饲料(1/30 w/w)。在术后第8天和第21天(仅80%组)处死大鼠，取胰腺称重，进行组织学分析。以假手术组大鼠相应部位体重归一化体重平均值为基础，计算胰腺残留物再生率。65%Sham组胰腺残留率为36.4±1.4%，80%Sham组为19.6±0.7%。P&lt；0.05时有统计学意义。在POD 8时，PX和Lig组的残余胰腺湿重在65%和80%的实验中均显著增加。正常饲料中Px/Lig/Sham的再生率分别为29±7/14±10/4±6和24±4/35±13/5±5%。饮食对其无明显影响。21d时，胰腺重量仅Px组显著增加。Px/Lig/Sham在正常饲料中的再生率为54±8/11±8/-13±4%。同样，饮食也没有显著影响。组织进行胰岛素免疫染色和PDX-1的原位杂交分析。在两种分析中，胰腺残留物没有显著差异。而在结扎胰管的LIG组，PDX-1在胰管上皮细胞中呈弥漫性表达，少数胰岛素阳性细胞表达，提示胰腺再生过程被激活。结论本研究未发现在饲料中添加胰酶的显著作用，提示肠道因素在胰腺再生中可能不起主要作用。POD 8和POD 21结果的明显差异是非常有趣的。对这种差异的原因的进一步研究似乎导致对胰腺再生机制的深刻理解较少。

项目成果

Sumi S, Kitamura Y, Endo S.: "Experimental studies in the influence of partial resection and duct ligature of the pancreas on regeneration of the rat pancreas"The Japanese Journal of Bilio-pancreatic Physiology. 18. 45-48 (2002)

Sumi S、Kitamura Y、Endo S.：“胰腺部分切除和导管结扎对大鼠胰腺再生影响的实验研究”日本胆胰生理学杂志。

角 昭一郎, 北村 義則, 遠藤 真一郎, 井上 一知: "膵切除と膵再生に関する研究"日本アフェレシス学会雑誌. 21. 141-144 (2002)

Shoichiro Kado、Yoshinori Kitamura、Shinichiro Endo、Kazutomo Inoue：“胰腺切除和胰腺再生的研究”日本血液成分协会杂志 21. 141-144 (2002)。

Sumi S, Kitamura Y, Endo S, Inoue K.: "Studies on pancreatectomy and pancreas regeneration. (in Japanese)"Japanese Journal of Apheresis. 21. 141-144 (2002)

Sumi S、Kitamura Y、Endo S、Inoue K.：“胰腺切除术和胰腺再生的研究。（日语）”日本血浆分离术杂志。

角 昭一郎, 北村 義則, 遠藤 真一郎: "ラット膵再生に対する膵管結紮と切除の影響に関する研究"胆膵の生理機能. 18. 45-48 (2002)

Shoichiro Kado、Yoshinori Kitamura、Shinichiro Endo：“胰管结扎和切除对大鼠胰腺再生的影响研究” Biliopancreatic Physiology 18. 45-48 (2002)。

Kitamura Y, Sumi S, Watanabe H, Urushihata T, Nio Y, Tamura K, Iwasaki S.: "Examination of glucose tolerance in the rat after total pancreatic duct ligation. (in Japanese)"The Japanese Journal of Bilio-pancreatic Physiology. 16. 61-64 (2000)

Kitamura Y、Sumi S、Watanabe H、Urushihata T、Nio Y、Tamura K、Iwasaki S.：“全胰管结扎后大鼠的葡萄糖耐量检查。（日语）”日本胆胰生理学杂志。