葛根素通过NF-κB通路抑制地塞米松诱导的hFOB1.19人成骨细胞凋亡  

Puerarin inhibits dexamethasone-induced apoptosis by hFOB1.19 human osteoblast cells through NF-κB pathway

作　　者：于冬冬[1] 王广斌[2] 赵丹阳[3] 

机构地区：[1]辽宁中医药大学附属医院骨一科,沈阳110032 [2]中国医科大学附属盛京医院,沈阳110004 [3]沈阳市第一人民医院,沈阳110044

出　　处：《中国骨质疏松杂志》2015年第8期929-933,共5页Chinese Journal of Osteoporosis

摘　　要：目的探索葛根素抑制地塞米松诱导的hFOB1.19人成骨细胞凋亡机制的实验研究。方法 MTS检测葛根素对hFOB1.19细胞增殖活性影响,免疫荧光检测DEX诱导h FOB1.19细胞凋亡及葛根素抑制DEX诱导的细胞凋亡的细胞核改变。Western blot检测p-p65、p-IκB的蛋白表达情况。Realtime PCR检测Caspase-3、Caspase-9的基因表达情况。ELISA检测加入NF-κB抑制PDTC后对凋亡的影响。结果 10-8M和10-9M葛根素明显增加细胞的增殖活性,10-5M和10-6M浓度的DEX作用72h后诱导细胞凋亡,加入10-8M葛根素后细胞凋亡受到抑制。Western blot结果显示,加入DEX后,p-NF-κB,pIκB的表达上调,DEX和PUE共同处理后,DEX诱导的NF-κB和IκB的磷酸化受到抑制。Realtime PCR显示DEX处理后caspase3、caspase8的mRNA基因表达明上调,而DEX和PUE共同作用后,caspase3、caspase8的mRNA基因表达下调。ELISA显示PUE抑制DEX诱导的细胞凋亡。PDTC、DEX、PUE共同处理后,PUE对DEX诱导的细胞的保护作用受到部分抑制。结论葛根素抑制地塞米松诱导的hFOB1.19细胞凋亡通过依赖于Caspase调节NF-κB通路。Objective To explore the mechanism of puerarin in inhibition of dexamethasone-induced apoptosis by h FOB1. 19 cells. Methods The effect of puerarin on cell proliferation of h FOB1. 19 cells was detected using MTS method. The effect of dexamethasone on apoptosis and the protect effect of puerarin on h FOB1. 19 cells were detected using immunefluorescence method.The protein expression p-p65 and p-IκB was detected using Western blotting. The gene expression of caspase-3 and caspase-9 was detected using real-time PCR. The effect of NF-κB inhibitor PDTC on cells apoptosis was detected using ELISA method. Results Puerarin of 10- 8Mand 10- 9Mconcentrations increased cell proliferation significantly. DEX of 10- 5Mand 10- 6Mconcentrations increased cell apoptosis after 72 h treatment. Puerarin of 10- 8Mconcentration inhibited DEX-induced cell apoptosis. Western blotting results showed that the protein expression of p-65 and p-IκB increased after DEX treatment. DEX-induced phosphorylation of NF-κB and IκB was inhibited after addition of PUE. Real-time PCR showed that the gene expression of caspase3 and caspase8 mRNA was up-regulated after DEX treatment. This expression was down-regulated after addition of PUE. ELISA results showed that PUE inhibited the cells apoptosis induced by DEX. When additioned with PDTC,DEX,and PUE,the protective effect of PUE was inhibited partially. Conclusion Puerarin inhibits dexamethasone-induced h FOB1. 19 cells apoptosis through caspase regulation of NF-κB pathway.

关 键 词：葛根素 地塞米松 成骨细胞 凋亡 

分 类 号：R861[医药卫生—临床医学]