Up-regulation of microglial cathepsin C expression and activity in lipopolysaccharide-induced neuroinflammation
1 Department of Anatomy, Dalian Medical University, No. 9, West Segment of South Lvshun Road, Dalian, Liaoning, 116044, China
2 Department of Physiology, Dalian Medical University, No. 9, West Segment of South Lvshun Road, Dalian, Liaoning, 116044, China
3 Graduate School, Dalian Medical University, No. 9, West Segment of South Lvshun Road, Dalian, Liaoning, 116044, China
4 General Surgery, Wafangdian Central Hospital, No. 3, Jinluan Road, Wafangdian, Liaoning, 116300, China
5 Department of Neurology, the First Affiliated Hospital of Dalian Medical University, No. 222, Zhongshan Road, Dalian, Liaoning, 116023, China
6 Department of Otolaryngology, the First Affiliated Hospital of Dalian Medical University, No. 222, Zhongshan Road, Dalian, Liaoning, 116023, China
Journal of Neuroinflammation 2012, 9:96 doi:10.1186/1742-2094-9-96Published: 20 May 2012
Cathepsin C (Cat C) functions as a central coordinator for activation of many serine proteases in inflammatory cells. It has been recognized that Cat C is responsible for neutrophil recruitment and production of chemokines and cytokines in many inflammatory diseases. However, Cat C expression and its functional role in the brain under normal conditions or in neuroinflammatory processes remain unclear. Our previous study showed that Cat C promoted the progress of brain demyelination in cuprizone-treated mice. The present study further investigated the Cat C expression and activity in lipopolysaccharide (LPS)-induced neuroinflammation in vivo and in vitro.
C57BL/6 J mice were intraperitoneally injected with either 0.9% saline or lipopolysaccharide (LPS, 5 mg/kg). Immunohistochemistry (IHC) and in situ hybridization (ISH) were used to analyze microglial activation, TNF-α, IL-1β, IL-6, iNOS mRNAs expressions and cellular localization of Cat C in the brain. Nitrite assay was used to examine microglial activation in vitro; RT-PCR and ELISA were used to determine the expression and release of Cat C. Cat C activity was analyzed by cellular Cat C assay kit. Data were evaluated for statistical significance with paired t test.
Cat C was predominantly expressed in hippocampal CA2 neurons in C57BL/6 J mice under normal conditions. Six hours after LPS injection, Cat C expression was detected in cerebral cortical neurons; whereas, twenty-four hours later, Cat C expression was captured in activated microglial cells throughout the entire brain. The duration of induced Cat C expression in neurons and in microglial cells was ten days and three days, respectively. In vitro, LPS, IL-1β and IL-6 treatments increased microglial Cat C expression in a dose-dependent manner and upregulated Cat C secretion and its activity.
Taken together, these data indicate that LPS and proinflammatory cytokines IL-1β, IL-6 induce the expression, release and upregulate enzymatic activity of Cat C in microglial cells. Further investigation is required to determine the functional role of Cat C in the progression of neuroinflammation, which may have implications for therapeutics for the prevention of neuroinflammation-involved neurological disorders in the future.