Chinese Journal of Tissue Engineering Research ›› 2015, Vol. 19 ›› Issue (11): 1712-1716.doi: 10.3969/j.issn.2095-4344.2015.11.014

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Calcium oxalate crystals stimulate expression of high mobility group box-1 protein in human macrophages  

Huang Peng, Deng Yao-liang, Li Cheng-yang, Tao Zhi-wei, Wang Xiang, Feng You-cai, Wu Bo   

  1. Department of Urinary Surgery, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China
  • Revised:2015-01-06 Online:2015-03-12 Published:2015-03-12
  • About author:Huang Peng, Master, Department of Urinary Surgery, First Affiliated Hospital of Guangxi Medical University, Nanning 530021, Guangxi Zhuang Autonomous Region, China

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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 cells, a large number of infiltrated monocytes and macrophages in the kidney will produce some inflammatory injuries and damage renal tubular epithelial cells, 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, real-time quantitative PCR was adopted to determine the mRNA expression of HMGB1. Tumor necrosis factor α and interleukin-6 levels in the cultured cell 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 gradually 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 cells had no changes within 0-12 hours, but significantly increased within 18-24 hours. ELISA results showed that the levels of tumor necrosis factor α and interleukin-6 in the cultured cell 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.


中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程


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Key words: High Mobility Group Proteins, Calcium Oxalate, Macrophages

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