免疫组化检测 Cerulein 诱导的急性胰损伤模型中小鼠和大鼠 Reg3γ 的表达。
Detection of Reg3γ by Immunohistochemistry in Cerulein-Induced Model of Acute Pancreatic Injury in Mice and Rats.
机构信息
From the Division of Applied Regulatory Science, Center for Drug Evaluation and Research, US Food and Drug Administration, Silver Spring, MD.
出版信息
Pancreas. 2019 Sep;48(8):1015-1025. doi: 10.1097/MPA.0000000000001382.
OBJECTIVE
In a continuation of previous work, Reg3γ protein was further evaluated as a biomarker of pancreatic injury using immunohistochemistry in an additional species.
METHODS
Mice and rats were treated with intraperitoneal cerulein injections, creating acute pancreatic injury. Mice received 2, 4, or 6 doses, and rats received 1, 2, or 3 doses of cerulein creating low, medium, and high treatment groups. Control animals were dosed with phosphate-buffered saline at corresponding volumes and intervals. Groups of 6 animals were killed 1, 3, 6, 24, and 48 hours after final treatments. Reg3γ immunohistochemical staining and image analysis were performed on pancreatic tissue obtained 6, 24, or 48 hours after control or cerulein treatment. Staining was quantified using image analysis software to calculate area of positivity as a percentage of total tissue area.
RESULTS
Percent positivity of Reg3γ in both species rose by 6 hours, peaked by 24 hours across all 3 cerulein doses, and dropped significantly by 48 hours. In high-dose rats with accompanying gene expression data, Reg3γ gene expression corresponded temporally with quantitative staining data.
CONCLUSIONS
Reg3γ staining quantified through image analysis showed a time- and dose-response in cerulein-treated mice and rats.
目的
在前一项工作的基础上，我们通过免疫组织化学法在另一种物种中进一步评估 Reg3γ 蛋白作为胰腺损伤生物标志物的作用。
方法
使用腹腔内给予亮蓝注射的方法对小鼠和大鼠进行处理，以造成急性胰腺损伤。小鼠接受 2、4 或 6 次剂量，大鼠接受 1、2 或 3 次亮蓝注射，形成低、中、高治疗组。对照动物按相应体积和间隔给予磷酸盐缓冲盐水。在末次治疗后 1、3、6、24 和 48 小时处死每组 6 只动物。在对照或亮蓝处理后 6、24 或 48 小时获得胰腺组织，进行 Reg3γ 免疫组织化学染色和图像分析。使用图像分析软件对染色进行定量，以计算阳性面积占总组织面积的百分比。
结果
两种物种的 Reg3γ 阳性百分比在 6 小时时升高，在所有 3 种亮蓝剂量下在 24 小时时达到峰值，并在 48 小时时显著下降。在伴有基因表达数据的高剂量大鼠中，Reg3γ 基因表达与定量染色数据在时间上相对应。
结论
通过图像分析定量的 Reg3γ 染色在亮蓝处理的小鼠和大鼠中显示出时间和剂量反应。
Zotero 插件