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目的动态观察血管紧张素II受体在胰腺纤维化大鼠胰腺组织中的表达并探讨其可能意义。方法采用胰管内注射2%三硝基苯磺酸(TNBS)诱导大鼠胰腺纤维化模型。制模后第3、21、28、35、42、49天分别处死大鼠,每组6只大鼠。对照组仅行剖腹术而未注射TNBS。应用VanGieson(V-G)染色观察胰腺组织纤维化程度。免疫组化和RT-PCR检测胰腺组织AT1蛋白、血管紧张素II受体mRNA和TNF-αmRNA表达。结果模型组大鼠胰腺组织细胞外基质合成较对照组明显增加。胰腺组织TNF-αmRNA表达逐渐增加并于第五周达峰值。AT1、AT2mRNA表达随时间不同呈不同程度的增加,分别于第28天、第35天达到峰值(382%和314%)。与AT1B受体mRNA相比,AT1受体亚型AT1AmRNA表达水平升高更为明显。免疫组化结果提示,模型组大鼠胰腺组织AT1蛋白表达增加并主要分布于纤维化区域。结论血管紧张素II可能通过AT1、AT2介导的途径参与了TNBS诱导的大鼠胰腺组织纤维化形成过程。
Objective To observe the expression of angiotensin II receptor in the pancreatic tissue of pancreatic fibrosis rats and to explore its possible significance. Methods Pancreatic fibrosis was induced by intraperitoneal injection of 2% trinitrobenzene sulfonic acid (TNBS) in rats. Rats were sacrificed on the 3rd, 21st, 28th, 35th, 42th and 49th days after the model was established. Six rats in each group. Control group only laparotomy without TNBS injection. Pancreatic tissue fibrosis was observed by Van Gieson (V-G) staining. The expression of AT1 protein, angiotensin II receptor mRNA and TNF-α mRNA in pancreatic tissue were detected by immunohistochemistry and RT-PCR. Results Compared with the control group, the extracellular matrix synthesis of pancreas in model group was significantly increased. The expression of TNF-αmRNA in pancreas gradually increased and peaked in the fifth week. The expression of AT1 and AT2 mRNA increased with time and reached the peak (382% and 314% respectively) on the 28th day and the 35th day respectively. Compared with AT1B receptor mRNA, AT1 receptor subtype AT1A mRNA expression increased more significantly. The results of immunohistochemistry suggested that the expression of AT1 protein in the pancreas tissue of the model group increased and mainly distributed in the fibrosis area. Conclusion Angiotensin II may participate in TNBS-induced fibrosis of rat pancreatic tissue through AT1 and AT2-mediated pathways.