Yan Liu1,2, Kai Zhao1, Zhe Xiong3, Yuanxiao Yi1, Chengliang Xiong1, Donghui Huang4, Hu Zhao5. 1. Institute of Reproduction Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. 2. Puren Hospital Affiliated to Wuhan University of Science and Technology, Wuhan, China. 3. Department of Physiology, Medical School of Jianghan University, Wuhan, China. 4. Institute of Reproduction Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China. drjhsyyjs@163.com. 5. Department of Human Anatomy, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
Abstract
INTRODUCTION: Urokinase plasminogen activator (uPA) is a serine protease and it also demonstrates proinflammatory properties. We thus hypothesized that uPA is involved in immunity of Sertoli cells in the rat testis. MATERIALS AND METHODS: The uPA gene(PLAU) promoter was cloned by RT-PCR and the transcriptional activation of core domain was screened by using Dual-Luciferase Reporter Assay System. The Sertoli cells were harvested from 20-day-old Sprague-Dawley male rats and total RNA was isolated. The uPA mRNA levels and MyD88 pathway were tested by qPCR. RESULTS: We successfully cloned the 1517-bp rat uPA gene and screened the core domain (-455/+40) in five different fragments of uPA promoter. The uPA expression and uPA promoter activity were upregulated in lipopolysaccharide (LPS)-stimulated Sertoli cells. Furthermore, the uPA expression was regulated through the MyD88-independent pathway by interdicting IRF3 and interferon b. CONCLUSION: uPA expression is likely induced by LPS through the core promoter domain of uPA. This finding implied that uPA played a role in the immune function of Sertoli cells in rat testis, which might provide the development of new treatments for male infertility.
INTRODUCTION: Urokinase plasminogen activator (uPA) is a serine protease and it also demonstrates proinflammatory properties. We thus hypothesized that uPA is involved in immunity of Sertoli cells in the rat testis. MATERIALS AND METHODS: The uPA gene(PLAU) promoter was cloned by RT-PCR and the transcriptional activation of core domain was screened by using Dual-Luciferase Reporter Assay System. The Sertoli cells were harvested from 20-day-old Sprague-Dawley male rats and total RNA was isolated. The uPA mRNA levels and MyD88 pathway were tested by qPCR. RESULTS: We successfully cloned the 1517-bp rat uPA gene and screened the core domain (-455/+40) in five different fragments of uPA promoter. The uPA expression and uPA promoter activity were upregulated in lipopolysaccharide (LPS)-stimulated Sertoli cells. Furthermore, the uPA expression was regulated through the MyD88-independent pathway by interdicting IRF3 and interferon b. CONCLUSION: uPA expression is likely induced by LPS through the core promoter domain of uPA. This finding implied that uPA played a role in the immune function of Sertoli cells in rat testis, which might provide the development of new treatments for male infertility.