大鼠和人外分泌胰腺中淀粉酶定位的比较免疫组织化学研究。

A comparative immunohistochemical study on amylase localization in the rat and human exocrine pancreas.

作者信息

Aughsteen A A

机构信息

Department of Anatomy and Histology, Faculty of Medicine, University of Jordan, Amman, Jordan.

出版信息

Saudi Med J. 2001 May;22(5):410-5.

PMID:11376382

Abstract

OBJECTIVE

To localize amylase enzyme immunohistochemically in the pancreatic acinar cells of rats and humans using polyclonal sheep anti-human amylase antibody, and to compare between the intensities of their amylase-immunostaining.

METHODS

Indirect immunofluorescence method was applied on formaldehyde-fixed, and paraffin-embedded pancreatic sections obtained from adult male Wistar rats and autopsied human samples. Primary incubation was performed using sheep anti-amylase antibody followed by secondary incubation with fluorescein isothiocyanate-labeled rabbit anti-sheep IgG serum. Control tests of amylase immunospecificity were also undertaken either by incubation with primary antibodies previously pre-adsorbed with an excess of human pancreatic amylase, or only with secondary antibodies.

RESULTS

The amylase immunofluorescence was positively and homogenously detected in all acinar cells of both rat and human pancreatic stained sections. The immunostaining was clearly demonstrated in the cell apices and peri-nuclear areas, but it was consistently brighter and more intense in the human acinar cells compared with that of the rat pancreas. Control tests of amylase immunofluorescence revealed the specificity of the antibodies applied for amylase localization in rat and human pancreas.

CONCLUSION

Although many previous immunohisto- and cytochemical reports have successfully localized amylase in the pancreas of different mammalian species, but all of them have used locally prepared anti-amylase antibodies. The present report successfully illustrates immuno-localization of amylase in the pancreatic acinar cells of rats and humans using commercial polyclonal sheep anti-human pancreatic amylase antibodies, and also suggests their useful application in the immunochemical studies on various mammalian species. Additionally, the results indicate a structural similarity between the human and rat pancreatic amylases, a concept required further exploration.

摘要

目的

使用多克隆羊抗人淀粉酶抗体，通过免疫组织化学方法在大鼠和人类胰腺腺泡细胞中定位淀粉酶，并比较它们淀粉酶免疫染色的强度。

方法

对取自成年雄性Wistar大鼠的甲醛固定、石蜡包埋的胰腺切片以及尸检人类样本应用间接免疫荧光法。先用羊抗淀粉酶抗体进行初次孵育，然后用异硫氰酸荧光素标记的兔抗羊IgG血清进行二次孵育。还通过与预先用过量人胰腺淀粉酶预吸附的一抗孵育，或仅与二抗孵育来进行淀粉酶免疫特异性的对照试验。

结果

在大鼠和人类胰腺染色切片的所有腺泡细胞中均阳性且均匀地检测到淀粉酶免疫荧光。免疫染色在细胞顶端和核周区域清晰可见，但与大鼠胰腺相比，人类腺泡细胞中的免疫染色始终更亮且更强。淀粉酶免疫荧光的对照试验揭示了所应用抗体在大鼠和人类胰腺中定位淀粉酶的特异性。

结论

尽管许多先前的免疫组织化学和细胞化学报告已成功地在不同哺乳动物物种的胰腺中定位了淀粉酶，但所有这些报告都使用了本地制备的抗淀粉酶抗体。本报告成功地展示了使用市售多克隆羊抗人胰腺淀粉酶抗体在大鼠和人类胰腺腺泡细胞中淀粉酶的免疫定位，并且还表明了它们在各种哺乳动物物种免疫化学研究中的有用应用。此外，结果表明人类和大鼠胰腺淀粉酶之间存在结构相似性，这一概念需要进一步探索。