·糖尿病基础研究·

　　【摘要】目的 研究SAA-siRNA沉默对波动性高糖培养下人肾小球系膜细胞(HMCs)炎症反应及细胞外基质(ECM)的影响，探讨波动性高糖与糖尿病慢性肾脏疾病(CKD)的关系及血清淀粉样蛋白A(SAA)在CKD发生发展中的作用。 方法 传代培养5~9代的HMCs，同步化后分6组进行实验。荧光倒置显微镜观察siRNA转染效率;real-time RT-PCR技术检测SAA及TNF-α的mRNA表达;Western Blot印迹检测细胞内SAA蛋白的表达;ELISA法检测细胞上清液中SAA、纤连蛋白(FN)及TNF-α的表达情况。 结果 分组培养48 h后，SAA、FN及TNF-α的表达高血糖(HG)组较对照(NC)组升高(P<0.05)，波动性高血糖(FG)组较HG组表达升高(P<0.05);波动性高血糖+SAA-siRNA(FG+SAA-siRNA)组SAA、FN及TNF-α表达较FG组降低(P<0.05)。 结论 波动性高血糖可引起HMCs更高水平SAA、FN及TNF-α的表达;通过SAA-siRNA沉默SAA表达，可减少波动性高血糖刺激下HMCs的FN、TNF-α的分泌，减少肾小球ECM积聚和炎症反应，为CKD防治提供新的实验依据。

　　【关键词】 血清淀粉样蛋白A;RNA干扰;人肾小球系膜细胞;波动性高血糖

The effect of serum amyloid A-siRNA silence on fluctuated high glucose induced secretion of extro-cellular matrix and inflammation factors in human mesangial cells FU Yu, WANG Qiu-yue, MA Dong-wei. Department of Endocrinology, The First Hospital, China Medical University, Shenyang 110001, China

　　【Abstract】 Objective To observe the effect of serum amyloid A-siRNA silence on the inflammatory response and extro-cellular matrix (ECM) in fluctuated high glucose induced human mesangial cells (HMCs), to explore the relationship between fluctuated high glucose and diabetic kidney disease (DKD), and to evaluate the role of SAA in the process of DKD. Methods Synchronized HMCs were divided into 6 groups. The transfection efficiency was observed with fluorescence microscope. The mRNA expression of SAA and tumor necrosis factor-α (TNF-α) were detected by real-time RT-PCR, the intro-cellular SAA protein was detected by Western Blot, and the protein expression of SAA, TNF-α and fibronectin (FN) were detected by ELISA. Results The expression of SAA, FN and TNF-α of the high glucose (HG) group was higher than that of the normal glucose group (P<0.05). The expression of the above factors of the fluctuated glucose (FG) group was higher than that of the HG group (P<0.05). The expression of the above factors of the FG+SAA-siRNA group was obviously lower than that of the FG group (P<0.05). Conclusion Fluctuated high glucose evokes more expression of SAA, FN and TNF-α compared with stable high glucose. Through the secretion of SAA-siRNA silence SAA, the secretion of FN and TNF-α in fluctuated high glucose-induced HMCs could be decreased and the accumulation of ECM and inflammation of HMCs could be decreased, providing a new experimental evidence for preventing DKD.

　　【Key words】Serum amyloid A (SAA); RNA interference; Human mesangial cells (HMCs); Fluctuated high glucose