小鼠胰腺对梯度胆囊收缩素的应答

doi:10.11958/20220309

天津医药 ›› 2022, Vol. 50 ›› Issue (10): 1056-1060.doi: 10.11958/20220309

小鼠胰腺对梯度胆囊收缩素的应答

武平¹(), 胡立娟², 徐晓晴¹, 李秋菊¹, 姚传山³, 王丰²^,^△()

1 天津医科大学研究生院（邮编300070）
2 天津医科大学附属南开医院急腹症器官损伤与中西医结合修复重点实验室
3 南开大学医学院

收稿日期:2022-03-03 修回日期:2022-06-17 出版日期:2022-10-15 发布日期:2022-10-20
通讯作者: 王丰 E-mail:pingwu@tmu.edu.cn;fengwangpi@tmu.edu.cn
作者简介:武平（1995）,男,硕士在读,主要从事胰腺生理和胰腺癌的基础方面研究。E-mail： pingwu@tmu.edu.cn

The response of mouse pancreas to gradient cholecystokinin

WU Ping¹(), HU Lijuan², XU Xiaoqing¹, LI Qiuju¹, YAO Chuanshan³, WANG Feng²^,^△()

1 The Graduate School, Tianjin Medical University, Tianjin 300070, China
2 The Laboratory of Acute Abdomen Disease Associated Organ Injury and Repair, Nankai Hospital Affiliated to Tianjin Medical University
3 Medical School, Nankai University

Received:2022-03-03 Revised:2022-06-17 Published:2022-10-15 Online:2022-10-20
Contact: WANG Feng E-mail:pingwu@tmu.edu.cn;fengwangpi@tmu.edu.cn

武平, 胡立娟, 徐晓晴, 李秋菊, 姚传山, 王丰. 小鼠胰腺对梯度胆囊收缩素的应答[J]. 天津医药, 2022, 50(10): 1056-1060.

WU Ping, HU Lijuan, XU Xiaoqing, LI Qiuju, YAO Chuanshan, WANG Feng. The response of mouse pancreas to gradient cholecystokinin[J]. Tianjin Medical Journal, 2022, 50(10): 1056-1060.

摘要/Abstract

摘要：

目的分析胆囊收缩素（CCK）对胰腺及胰腺癌发生的影响,并探讨绿茶成分表没食子儿茶素没食子酸酯（EGCG）的抗癌作用。方法将生长期小鼠随机分为4组,分别给予含大豆胰蛋白酶抑制剂（STI）0 g/L（11只）、2 g/L（11只）、4 g/L（10只）和8 g/L（11只）的饮水,2周后处死小鼠,记录胰腺湿质量。采用酶联免疫吸附试验（ELISA）检测血浆CCK水平;测定胰腺组织中蛋白质、DNA、胰蛋白酶和脂肪酶含量;免疫组织化学染色检测胰腺组织增殖细胞核抗原（PCNA）阳性细胞百分比。采用胰腺内包埋化学致癌物二甲基苯并蒽（DMBA）法建立癌发生模型。将15只小鼠随机均分为正常对照组、DMBA包埋组、DMBA+2 g/L STI组、DMBA+8 g/L STI组、DMBA+8 g/L STI+EGCG组,6周后处死小鼠,取胰腺进行组织学分析。结果 2 g/L STI组CCK水平高于0 g/L STI组,8 g/L STI组高于0、2和4 g/L STI组（P<0.05）。2、4 g/L STI组胰腺湿质量高于0 g/L STI组,8 g/L STI组高于0、2 g/L STI组（P<0.05）。2、4 g/L STI组胰腺蛋白质和DNA含量高于0 g/L STI组,8 g/L STI组高于0、2和4 g/L STI组（P<0.05）。0、2、4和8 g/L STI组胰蛋白酶含量依次升高;2、4、8 g/L STI组胰脂酶含量高于0 g/L STI组（P<0.05）。0、2、4、8 g/L STI组小鼠胰腺组织PCNA阳性细胞比例依次升高（P<0.05）。胰腺癌前病变程度随饮水中的STI剂量增大而加重,EGCG可减轻此病变。结论 STI可诱发梯度CCK血症,并刺激胰腺增生、胰酶生成及胰腺癌发生,而EGCG具有减轻胰腺癌前病变的作用。

关键词: 大豆胰蛋白酶抑制剂, 胆囊收缩素, 胰腺生长, 胰蛋白酶, 胰脂肪酶, 表没食子儿茶素没食子酸酯

Abstract:

Objective To investigate the effect of cholecystokinin (CCK) on pancreas and occurrence of pancreatic cancer, and to explore the anticancer effect of a component of green tea, epigallocatechin galate (EGCG). Methods Growing mice were randomly divided into 4 groups, and mice were given water containing soybean trypsin inhibitor (STI) 0 g/L (n=11), 2 g/L (n=11), 4 g/L (n=10) and 8 g/L (n=11), respectively. After 2 weeks, mice were sacrificed and the wet weight of pancreas was recorded. Plasma CCK level was detected by enzyme-linked immunosorbent assay (ELISA). The contents of protein, DNA, trypsin and lipase in pancreatic tissue were determined. The percentage of proliferating cell nuclear antigen (PCNA) positive cells in pancreatic tissue was detected by immunohistochemical staining. The carcinogenesis model was established by embedding dimethyl benzoanthracene (DMBA) in pancreas. Fifteen mice were randomly divided into the normal control group, the DMBA embedding group, the DMBA+2 g/L STI group, the DMBA+8 g/L STI group and the DMBA+8 g/L STI+EGCG group. After 6 weeks, the mice were sacrificed and the pancreas was taken for histological analysis. Results CCK level was higher in the 2 g/L STI group than that in the 0 g/L STI group, and which was higher in the 8 g/L STI group than that in the 0, 2 and 4 g/L STI group (P<0.05). The wet weight of pancreas was higher in the 2 g/L STI group and the 4 g/L STI group than that in the 0 g/L STI group, and which was higher in the 8 g/L STI group than that in the 0 g/L STI group and the 2 g/L STI group (P<0.05). The contents of pancreatic protein and DNA were higher in the 2 g/L STI group and 4 g/L STI group than those in the 0 g/L STI group, and which were higher in the 8 g/L STI group than those in the 0 g/L STI group, 2 g/L STI group and 4 g/L STI group (P<0.05). The content of trypsin increased successively in the 0 g/L STI group, the 2 g/L STI group, the 4 g/L STI group and the 8 g/L STI group. The lipase content was significantly higher in the 2 g/L STI group, the 4 g/L STI group and the 8 g/L STI group than that in the 0 g/L STI group (P < 0.05). The proportion of PCNA-positive cells in pancreatic tissue of mice increased successively in the 0 g/L STI group, 2 g/L STI group, the 4 g/L STI group and the 8 g/L STI group (P < 0.05). The degree of pancreatic cancer prelesion was aggravated with the increase of STI dose in drinking water, and EGCG can alleviate this lesion. Conclusion STI can induce gradient cholecystokininemia, stimulate pancreatic hyperplasia, trypsin production and pancreatic cancer, while EGCG can alleviate pancreatic precancerous lesions.

Key words: Soybean trypsin inhibitor, cholecystokinin, pancreatic growth, trypsin, lipase, Epigallocatechin gallate

中图分类号:

R335.9

图/表 5

表1 各组小鼠CCK水平及胰腺湿质量比较 ($\bar{x} \pm s$)

Tab. 1 Comparison of CCK concentration and pancreatic wet mass of mice between the four groups

组别	n	CCK（ng/L）	胰腺湿质量（mg）
0 g/L STI组	11	57.33±15.23	152.64±17.74
2 g/L STI组	11	79.08±13.13^a	185.73±31.92^a
4 g/L STI组	10	67.06±9.62	198.80±29.02^a
8 g/L STI组	11	92.84±9.25^abc	220.45±43.66^ab
F		17.512^＊＊	8.619^＊＊

表2 各组小鼠胰腺蛋白质和DNA含量比较 ($\bar{x} \pm s$)

Tab. 2 Comparison of protein and DNA content in pancreas of mice between the four groups

组别	n	蛋白质含量（mg）	DNA含量（μg）
0 g/L STI组	11	14.30±2.37	228.29±27.03
2 g/L STI组	11	19.70±4.72^a	281.87±35.36^a
4 g/L STI组	10	20.63±4.49^a	299.26±34.12^a
8 g/L STI组	11	25.85±5.68^abc	346.62±32.88^abc
F		12.252^＊＊	24.892^＊＊

3 各组小鼠胰腺组织中消化酶含量比较 ($\bar{x} \pm s$)

Tab. 3 Comparison of digestive enzymes of pancreatic tissue of mice between the four groups

组别	n	胰蛋白酶含量（U/mg）	胰脂酶含量（ng/g）
0 g/L STI组	11	5.53±0.62	325.89±63.38
2 g/L STI组	11	6.86±0.57^a	435.03±35.89^a
4 g/L STI组	10	7.98±0.57^ab	468.73±82.43^a
8 g/L STI组	11	9.74±0.88^abc	469.35±96.56^a
F		76.520^＊＊	9.391^＊＊

图1 小鼠胰腺组织PCNA免疫组织化学染色结果（比例尺=25 μm）箭头所指为典型PCNA阳性染色细胞。

Fig.1 Results of immunohistochemical staining of PCNA in pancreas (scale = 25 μm)

图2 CCK和EGCG对胰腺包埋DMBA小鼠癌发生的作用（比例尺=25 μm） A~E分别为正常对照组、DMBA包埋组、DMBA+2 g/L STI组、DMBA+8 g/L STI组、DMBA+8 g/L STI+EGCG组;箭头所指为具有代表性的结构。

Fig.2 Effects of CCK and EGCG on neoplastic lesion in mouse pancreas embedded with DMBA (scale = 25 μm)

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小鼠胰腺对梯度胆囊收缩素的应答

The response of mouse pancreas to gradient cholecystokinin

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