晚期糖化终产物对β-淀粉样蛋白诱导PC12细胞凋亡作用机制的实验研究

Experimental study on the mechanism of the apoptotic effect of advanced glycation end products on PC12 cell induced by β-amyloid protein

目的研究AGEs对Aβ诱导的PC12细胞的作用机制是否与氧化应激有关,阻断RAGE活性能否作为有效的作用靶点。方法Aβ诱导PC12细胞后加入低、中、高浓度AGEs孵育12h;胰蛋白酶处理PC12细胞后加入Aβ,MTT法检测细胞活力,流式细胞仪测定细胞凋亡,RTPCR法测定RAGE,NFκB的表达。结果MTT法显示随AGEs浓度升高,细胞活力下降,胰蛋白酶处理组细胞活力升高,流式细胞仪检测显示随AGEs浓度增加,细胞凋亡率增加,RTPCR法半定量分析RAGE、NFκB表达随AGEs浓度升高而增高。结论AGEs对Aβ诱导的PC12细胞神经毒性损害由RAGE介导,并与RAGE引起NFκB表达有关,阻断RAGE可减低其神经毒性,阻断RAGE活性可作为有效的药物作用靶点。

Objective To explore whether the mechanism of the effect of advanced glycation end products (AGE) on PC12 cell induced by β-amyloid protein relates to oxidation stress and to study whether inhibiting the activity of RAGE is a benefit drug target. Methods PC-12 cells were treated with Aβ_~25-35 , then AGEs with different concentration were added to the culture medium for 12 hours. The cell activity was measured by MTT assay, the apoptosis situation was detected by flowcytometry and the expressions of RAGE and NF-κB were measured by RT-PCR assay. Results Cell activity decreased with the increase of AGEs concentration, but the activity increased after treatment with pancreatic protein enzyme. The cell apoptosis rate and the expressions of RAGE and NF-κB were elevated with AGE concentration increasing. Conclusions RAGE triggers the neural toxicity of RAGE to the PC-12 cell induced by Aβ, and the molecular mechanism is connected with the expression of NF-κB induced by RAGE. Inhibiting the activity of RAGE can be an effective drug target.