摘要
目的:研究大黄素脂质纳米微泡对机械牵张引发的Ⅱ型肺泡上皮细胞MAPK信号通路和炎性介质的影响和作用机制。方法:以卵磷脂和PVP为载体材料,采用静电纺丝技术制备纳米纤维膜,填充全氟丙烷,通过自组装技术形成大黄素脂质纳米微泡。分离纯化原代大鼠AT-Ⅱ细胞,应用Western-Blot和ELISA方法观察AT-Ⅱ细胞在20%机械牵张应力的4、8、16、24、48h刺激作用下,p-P38/P38、p-ERK/ERK、p-JNK/JNK蛋白表达和TNF-α、IL-1β、IL-6分泌水平的动态变化;及进一步将大黄素脂质纳米微泡加入AT-Ⅱ细胞对VILI进行干预,并检测干预效果。结果:牵张刺激后,AT-Ⅱ细胞p-P38、p-ERK、p-JNK蛋白表达的水平显著升高,并在4～16h范围内表达水平持续升高,但p38、ERK、JNK蛋白表达无变化;AT-Ⅱ细胞TNF-α、IL-1β、IL-6分泌水平在8h内均无变化,而在随后16h的牵张刺激逐渐显著升高。在VILI刺激下,加入大黄素脂质纳米微泡进行干预,均可以不同程度下调p-P38、p-ERK、p-JNK蛋白表达和抑制TNF-α、IL-1β、IL-6的释放水平。结论:大黄素脂质纳米微泡对机械所致肺损伤(VILI)有保护作用,其机制可能与下调MAPK信号通路蛋白表达来调节炎性介质释放有关。
Objective：To investigate the effect of emodin lipid nano-microbubble on MAPK signaling pathway and the level of inflammation cytokine in AT-Ⅱ cells induced by mechanical stretch and its mechanism. Methods： Emodin nanofibers, prepared by electrospinning with lecithin and PVP as carrier, were put into a seal bottle full of perfluoropropane, after they mixed with water and turned into self-assembled nano-microbubble. AT-Ⅱ cells, separated and purified from primary rat AT-Ⅱ cells, suffered 20% mechanical stretch for 4,8, 16,24,48 b, and its protein expressions of p-P38/P38, p-ERK/ERK, p-JNK/JNK and the inflammation cytokine release levels of TNF-α, IL-1β, IL-6 were detected by Western-Blot and ELISA. And further oberved the intervention effect of emodin lipid nano-microbubble on VILI. Results： The protein expressions of p-P38, p-ERK ,p-JNK were significantly increased in AT-Ⅱ cells induced by mechanical stretch, and continuously evaluated from the 4 - 16 h, but the protein expressions of p38, ERK, JNK had no significant difference. The release levels of TNF-α, IL-1β, IL-6 in AT-Ⅱ ceils had no change during 8 h, and they were gradually increased significantly in followed 16 h. In AT-Ⅱ cells induced by mechanical stretch,due to intervention effect of emodin lipid nano- microbubble, the the protein expressions of p-P38, p-ERK, p-JNK and the inflammation cytokine release levels of TNF-α, IL-1β, IL-6 were significantly decreased. Conclusion ： Emodin lipid nano-microbubble shows protective effect on AT-Ⅱ cells induced by mechanical stretch （VILI） ,and its mechanism may be related to down-regulation of protein expression of MAPK signaling pathway to regulate the release levels of inflammatory cytokine.
出处
《中药材》
CAS
CSCD
北大核心
2013年第6期967-971,共5页
Journal of Chinese Medicinal Materials
基金
广东省自然科学基金(S2012010010157)
关键词
大黄素脂质纳米微泡
机械牵张
Ⅱ型肺泡上皮细胞
MAPK信号通路
炎性介质
Emodin lipid nano-microbubble
Mechanical stretch
Type Ⅱ alveolar epithelial cells （ AT-Ⅱ cells）
MAPK signalpathway
Inflammation cytokine
