中国组织工程研究Issue(11)：1712-1716,5.DOI:10.3969/j.issn.2095-4344.2015.11.014
草酸钙晶体刺激下人巨噬细胞高迁移率族蛋白B1的表达
Calcium oxalate crystals stimulate expression of high mobility group box-1 protein in human macrophages
黄鹏 1邓耀良 1黎承杨 1陶芝伟 1王翔 1奉有才 1吴博1
作者信息
- 1. 广西医科大学第一附属医院泌尿外科，广西壮族自治区南宁市 530021
- 折叠
摘要
Abstract
BACKGROUND:There are a large amount of infiltrated monocytes and macrophages around the renal interstitial crystals in a kidney model, indicating that macrophages may be involved in the process of crystal deposition in the kidney. As macrophages are important human innate immune cels, a large number of infiltrated monocytes and macrophages in the kidney wil produce some inflammatory injuries and damage renal tubular epithelial cels, which ultimately lead to the formation of stones. OBJECTIVE: To study the expression of after high mobility group box-1 protein (HMGB1) in macrophages stimulated by calcium oxalate monohydrate crystals. METHODS:Western blot assay was used to detect total protein and HMGB1 levels in the cytoplasm of macrophages under 100 mg/L calcium oxalate monohydrate stimulation for 0, 6, 12, 24, 36 hours. Meanwhile,
<br> real-time quantitative PCR was adopted to determine the mRNA expression of HMGB1. Tumor necrosis factor α and interleukin-6 levels in the cultured cel supernatant at 0, 1, 2 and 4 hours after calcium oxalate monohydrate stimulation were measured using ELISA. RESULTS AND CONCLUSION: Under the calcium oxalate monohydrate stimulation, the level of HMGB1 in the cytoplasm of macrophages was lower within 0-6 hours, but gradualy increased within 12-36 hours; the level of HMGB1 in the total protein of macrophages was not high within 0-6 hours, began to increase at 12 hours, and then kept at a higher level within 24-36 hours. RT-PCR results showed that under the calcium oxalate monohydrate stimulation, the mRNA level of HMGB1 in the cultured cels had no changes within 0-12 hours, but significantly increased within 18-24 hours. ELISA results showed that thelevels of tumor necrosis factor α and interleukin-6 in the cultured cel supernatant subject to calcium oxalate monohydrate were increased at 2 hours and peaked at 4 hours. These findings indicate that calcium oxalate monohydrate can increase the HMGB1 expression in macrophages at mRNA and protein levels, and also induce the increase in tumor necrosis factor α and interleukin-6 expression in macrophages. Moreover, the expression of HMGB1 is later than the release of tumor necrosis factor α and interleukin-6.关键词
组织构建/组织工程/高迁移率族B1/一水草酸钙/巨噬细胞Key words
High Mobility Group Proteins/Calcium Oxalate/Macrophages分类
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黄鹏,邓耀良,黎承杨,陶芝伟,王翔,奉有才,吴博..草酸钙晶体刺激下人巨噬细胞高迁移率族蛋白B1的表达[J].中国组织工程研究,2015,(11):1712-1716,5.
