中国骨质疏松杂志
中國骨質疏鬆雜誌
중국골질소송잡지
Chinese Journal of Osteoporosis
2015年
8期
929-933
,共5页
葛根素%地塞米松%成骨细胞%凋亡
葛根素%地塞米鬆%成骨細胞%凋亡
갈근소%지새미송%성골세포%조망
Puerarin%Dexamethasone%Osteoblast cells%Apoptosis
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hFOB1.19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 -8 M和10 -9 M葛根素明显增加细胞的增殖活性,10 -5 M和10 -6 M浓度的DEX作用72h后诱导细胞凋亡,加入10 -8 M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,p-IκB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.
目的 探索葛根素抑製地塞米鬆誘導的hFOB1.19 人成骨細胞凋亡機製的實驗研究. 方法 MTS檢測葛根素對hFOB1.19細胞增殖活性影響,免疫熒光檢測DEX誘導hFOB1.19細胞凋亡及葛根素抑製DEX誘導的細胞凋亡的細胞覈改變. Western blot檢測p-p65、p-IκB的蛋白錶達情況. Realtime PCR檢測Caspase-3、Caspase-9的基因錶達情況. ELISA檢測加入NF-κB抑製PDTC後對凋亡的影響. 結果 10 -8 M和10 -9 M葛根素明顯增加細胞的增殖活性,10 -5 M和10 -6 M濃度的DEX作用72h後誘導細胞凋亡,加入10 -8 M葛根素後細胞凋亡受到抑製. Western blot結果顯示,加入DEX後,p-NF-κB,p-IκB的錶達上調,DEX和PUE共同處理後, DEX誘導的NF-κB和IκB的燐痠化受到抑製. Realtime PCR顯示DEX處理後caspase3、caspase8的mRNA基因錶達明上調,而DEX和PUE共同作用後,caspase3、caspase8的mRNA基因錶達下調. ELISA顯示PUE抑製DEX誘導的細胞凋亡. PDTC、DEX、PUE共同處理後,PUE對DEX誘導的細胞的保護作用受到部分抑製. 結論 葛根素抑製地塞米鬆誘導的hFOB1.19細胞凋亡通過依賴于Caspase調節NF-κB通路.
목적 탐색갈근소억제지새미송유도적hFOB1.19 인성골세포조망궤제적실험연구. 방법 MTS검측갈근소대hFOB1.19세포증식활성영향,면역형광검측DEX유도hFOB1.19세포조망급갈근소억제DEX유도적세포조망적세포핵개변. Western blot검측p-p65、p-IκB적단백표체정황. Realtime PCR검측Caspase-3、Caspase-9적기인표체정황. ELISA검측가입NF-κB억제PDTC후대조망적영향. 결과 10 -8 M화10 -9 M갈근소명현증가세포적증식활성,10 -5 M화10 -6 M농도적DEX작용72h후유도세포조망,가입10 -8 M갈근소후세포조망수도억제. Western blot결과현시,가입DEX후,p-NF-κB,p-IκB적표체상조,DEX화PUE공동처리후, DEX유도적NF-κB화IκB적린산화수도억제. Realtime PCR현시DEX처리후caspase3、caspase8적mRNA기인표체명상조,이DEX화PUE공동작용후,caspase3、caspase8적mRNA기인표체하조. ELISA현시PUE억제DEX유도적세포조망. PDTC、DEX、PUE공동처리후,PUE대DEX유도적세포적보호작용수도부분억제. 결론 갈근소억제지새미송유도적hFOB1.19세포조망통과의뢰우Caspase조절NF-κB통로.
Objective To explore the mechanism of puerarin in inhibition of dexamethasone-induced apoptosis by hFOB1.19 cells.Methods The effect of puerarin on cell proliferation of hFOB1.19 cells was detected using MTS method.The effect of dexamethasone on apoptosis and the protect effect of puerarin on hFOB1.19 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 -8 M and 10 -9 M concentrations increased cell proliferation significantly.DEX of 10 -5 M and 10-6 M concentrations increased cell apoptosis after 72h treatment.Puerarin of 10 -8 M concentration 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 hFOB1.19 cells apoptosis through caspase regulation of NF-κB pathway.
