Inhibitory effect of colchicine on transforming growth factor β1/Smads pathway in rat models of chronic pancreatitis

doi:10.3969/j.issn.2095-4344.2014.49.023

Abstract

Abstract:

BACKGROUND: Pancreatic stellate cells transforming growth factor β1/Smads signaling pathway activation is probably a main molecular mechanism of pancreatic fibrosis. If this pathway can be blocked, the progression of fibrosis of tissues with chronic pancreatitis will be inhibited.

OBJECTIVE: To study the inhibitory effect of colchicine on transforming growth factor β1/Smads pathway in chronic pancreatitis rat models.

METHODS: Healthy male Sprague-Dawley rats were randomly divided into colchicines-treated group and chronic pancreatitis group. After successful establishment of rat models of chronic pancreatitis, the rats in the colchicines-treated group were intraperitoneally injected with colchicine 150 μg/kg daily. The rats in the chronic pancreatitis group were intraperitoneally injected with equal volume of physiological saline daily. Pancreatic tissues were collected after 3 months. Hematoxylin-eosin staining was used to observe histopathological changes of pancreatic tissue. Immunohistochemical staining was used to detect the expressions of transforming growth factor β1 in pancreatic tissue. Western blot assay was utilized to detect the expressions of P-Smad2, P-Smad3 and α-SMA protein in pancreatic stellate cells.

RESULTS AND CONCLUSION: Hematoxylin-eosin staining results revealed that compared with the colchicines-treated group, glandular tissue had reduced, while fibrous connective tissue and inflammatory cells had increased obviously and replaced the pancreatic gland tissue in the chronic pancreatitis group. Immunohistochemical staining results demonstrated that the expression levels of transforming growth factor β1 and the index of positive cells were significantly lower in the colchicines-treated group than those in the chronic pancreatitis group (P < 0.05). Western blot assay results revealed that the results of P-Smad2/β-actin, P-Smad3/β-actin and α-SMA/β-actin in pancreatic stellate cells were significantly lower in the chronic pancreatitis group than those in the colchicines-treated group (P < 0.05). Results suggested that colchicine could inhibit the activity of transforming growth factor β1/Smads pathway and pancreatic tissue fibrosis in chronic pancreatitis rats. Therefore, colchicine can be used as a new candidate therapeutic scheme for chronic pancreatitis fibrosis.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

全文链接：

Key words: transforming growth factor beta 1, smad2 protein, smad3 protein, pancreatitis, chronic

CLC Number:

R318

Lu Hong-wei, Zhang Ya-fei, Ji Hong, Wang Jin-long, Li Yi-ming. Inhibitory effect of colchicine on transforming growth factor β1/Smads pathway in rat models of chronic pancreatitis [J]. Chinese Journal of Tissue Engineering Research, 2014, 18(49): 8001-8006.

Figures/Tables 3

2.1 实验动物数量分析选取20只SD大鼠，分为慢性胰腺炎组及秋水仙碱干预组，每组10只，造模成功，实验过程无动物脱失，全部进入结果分析。 2.2 慢性胰腺炎大鼠模型的建立及秋水仙碱对慢性胰腺炎大鼠胰腺组织内纤维化的抑制作用实验后3个月取大鼠胰腺组织制成冰冻切片，进行病理学观察。慢性胰腺炎组胰腺组织内腺体组织减少，而纤维结缔组织、炎细胞明显增多并取代胰腺腺体组织；而秋水仙碱干预组胰腺组织内腺体基本正常，但腺体间有纤维结缔组织增生及炎细胞浸润显著少于慢性胰腺炎组(图1)。 2.3 秋水仙碱能够抑制慢性胰腺炎大鼠胰腺组织内TGF-β1的表达秋水仙碱干预组TGF-β1阳性表达级别为+，慢性胰腺炎组为+++；秋水仙碱干预组TGF-β1表达阳性细胞指数为11.92±6.57，慢性胰腺炎组为60.53±5.57。秋水仙碱干预组TGF-β1阳性表达级别和阳性细胞指数显著低于慢性胰腺炎组，两组比较差异有显著性意义(P < 0.05)，见图2。 2.4 秋水仙碱能够抑制慢性胰腺炎大鼠胰腺组织内P-Smad2、P-Smad3的表达两组均在实验后3个月时取材，Western blot法测定胰腺组织中的P-Smad2、P-Smad3及β-actin，将所拍照片结果应用Image Pro图像分析软件处理后，分别得出其积分吸光度值，然后分别计算出两组P-Smad2、P-Smad3与β-actin积分吸光度之比值，数据经统计学处理并进行比较。结果所示，术后3个月时慢性胰腺炎组胰腺组织内P-Smad2/β-actin、P-Smad3/β-actin达到了1.02±0.38和0.87±0.26，而秋水仙碱干预组则分别为0.21±0.11和0.34±0.14，两组间差异有显著性意义(P < 0.05)，见图3。 2.5 秋水仙碱能够抑制慢性胰腺炎大鼠胰腺组织内α-SMA的表达 Western blot法测定胰腺组织中的α-SMA及β-actin，将所拍照片结果应用Image Pro图像分析软件处理后，分别得出其积分吸光度值，然后分别计算出两组α-SMA与β-actin积分吸光度之比值，数据经统计学处理并进行比较。结果所示，术后3个月时慢性胰腺炎组胰腺组织内α-SMA/β-actin达到了1.86±0.73，而秋水仙碱干预组则为0.42±0.15，两组间差异有显著性意义(P < 0.05)，见图4。"

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