TRPC对脂多糖诱导的星形胶质细胞TNF-α和NO分泌的影响

目的 探讨TRPC对脂多糖诱导的星形胶质细胞TNF-α和NO分泌的影响.方法 通过摇床筛选法纯化大鼠大脑皮层星形胶质细胞,用免疫荧光法鉴定其纯度.细胞培养至80%左右融合时加入0.5 μg/mL 脂多糖(LPS)后用维持液分别培养0、2、6、12、24和48 h,检测TNF-α和NO分泌情况,观察TRPC阻断剂 2-A...

Effects of TRPC on LPS induced secretion of TNF-α and NO in cultured cortical Astrocytes

【Abstract】 Objective To investigate the effects of the TRPC on the secretion of TNF-α and NO from astrocytes stimulated by LPS. Methods The astrocytes were isolated and purified by shaking the flasks in a horizontal orbital shaker. Purified astrocytes were identified by immunofluorescence. While cell fusion ratio arrived 80%, cells were exposed to 0.5μg/mL LPS for 0 h, 2 h, 6 h, 12 h, 24 h, 48 h, or treated with 10μM 2-APB and 5μM SKF96365 simultaneously. TNF-α and NO concentration in cell culture supernatants were measured. Results The production of TNF-α, induced by LPS for 2-48 h were significantly increased (p<.0.01), while the NO secretion stimulated by LPS for 24-48 h were significantly increased (p<0.01). Treated with TRPC blockers 2-APB and SKF96365 both partially inhibited the secretion of TNF-α and NO from astrocytes (p<0.01). Conclusion The inhibition of TRPC channels could reduced the secretion of TNF-α and NO induced by LPS. Our results suggest that TRPC channel may act to regulate astrocytes activation in the pathophysiology of brain inflammation diseases.