| 沈书旭.幽门螺杆菌通过NF-κB信号通路调控胃上皮细胞自噬及凋亡的机制研究[J].安徽医药,2018,22(6):1058-1063. |
| 幽门螺杆菌通过NF-κB信号通路调控胃上皮细胞自噬及凋亡的机制研究 |
| Regulation of autophagy and apoptosis of gastric epithelial cells by helicobacter pylori through NF-κB signaling pathway |
| 投稿时间：2016-10-08 |
| DOI： |
| 中文关键词: 幽门螺杆菌 上皮细胞 自噬 细胞凋亡 NF-κB |
| 英文关键词: Helicobacter Epithelial cells Autophagy Apoptosis NF-kappa B |
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| 中文摘要: |
| 目的 研究幽门螺杆菌通过NF-κB信号通路调控胃上皮细胞自噬及凋亡的机制。 方法 体外培养幽门螺杆菌(Helicobacter pylori,Hp)及人胃上皮AGS细胞株,收集Hp处理0 h、3 h、6 h、9 h及对照组、18 μmol·L-1 NF-κB特异性抑制剂SN50处理6 h、18 μmol·L-1 SN50+Hp处理6 h的AGS细胞,利用流式细胞仪分别检测各组细胞的自噬率及凋亡率；收集Hp梯度时间处理后的AGS细胞,提取各组总蛋白,蛋白质印迹法(Western Blot)检测自噬相关蛋白Beclin1、微管相关蛋白1轻链3Ⅰ(LC3Ⅰ)、微管相关蛋白1轻链3Ⅱ(LC3Ⅱ),凋亡相关蛋白B细胞淋巴瘤/白血病-2(Bcl-2)、Bcl-2相关X蛋白(Bax)及NF-κB通路相关蛋白p65、p-p65的表达。 结果 AGS细胞的自噬率与凋亡率均随着Hp作用时间的延长逐渐升高,各时间点与0 h相比均差异有统计学意义(P<0.01)。自噬相关蛋白Beclin1、LC3Ⅱ、凋亡相关蛋白Bax及NF-κB通路相关蛋白p-p65的表达随着Hp作用时间的延长逐渐增高,自噬相关蛋白LC3Ⅰ及凋亡相关蛋白Bcl-2蛋白表达随着Hp作用时间的延长逐渐降低,与0 h相比,各检测时间点蛋白表达差异有统计学意义(P<0.01)。NF-κB通路相关蛋白p65的表达未出现显著性变化(P>0.05)。与对照组相比[(6.56±1.40)%、(11.76±1.70)%],Hp处理后,AGS细胞自噬率(9.86±1.78)%与凋亡率(13.25±1.56)%显著增强(t1=11.054,P1=0.000;t2=10.456,P2=0.000);SN50处理后,AGS细胞自噬率(19.65±2.14)%与凋亡率(28.65±2.14)%显著减弱(t1=3.403,P1=0.009;t2=2.799,P2=0.023);Hp与SN50联合处理后,AGS细胞自噬率(20.65±2.14)%与凋亡率(29.65±2.14)%显著高于对照组,但低于Hp单独处理组,差异有统计学意义(t1=5.164,P1=0.001;t2=5.890,P2=0.000;t3=4.718,P3=0.002;t4=5.738,P4=0.000)。 结论 Hp刺激显著促进AGS细胞的自噬与凋亡,作用机制可能与NF-κB通路的活化有关。 |
| 英文摘要: |
| Objective To investigate the mechanism of the regulation of autophagy and apoptosis of gastric epithelial cells by helicobacter pylori (Hp) through NF-κB signaling pathway. Methods Hp and human gastric epithelial cell lines AGS were cultivated in vitro,AGS cells were assigned into the control group,Hp treatment 0 h,3 h,6 h,9 h,18 μmol·L-1 SN50 treatment 6 h,18 μmol·L-1 SN50+Hp treatment 6 h.Flow cytometry was used to detect autophagy and apoptosis of cells in each group.AGS cells treated with Hp gradient time were collected and total protein was extracted from each group.Western blot was used to detect the expression of autophagy-related protein Beclin 1,microtubule-associated protein 1 light chain 3Ⅰ (LC3Ⅰ),microtubule-associated protein 1 light chain 3Ⅱ (LC3Ⅱ),apoptosis related protein Bcell lymphoma/lewkmia-2 (Bcl-2),Bcl-2 associated Xprotein (Bax) and NF-κB pathway related proteins p65,p-p65. Results AGS cells autophagy and apoptosis rate increased gradually with prolonged duration of action of Hp;Hp treatment 3 h,6 h,9 h had significant differences compared with 0 h (P<0.01).The expression of autophagy-related protein Beclin1,LC3Ⅱ,apoptosis-related protein Bax and NF-κB pathway-associated protein p-p65 were gradually increased with Hp treatment prolongation.The expression of autophagy-related protein LC3Ⅰ and apoptosis-related protein Bcl-2 protein expression were gradually decreased with Hp treatment prolongation,the protein expressions at 3 h,6 h,9 h were significantly different compared with 0 h (P<0.01).The expression of NF-κB pathway-associated protein p65 had no significant change (P>0.05).Compared with the control group[(6.56±1.40)%,(11.76±1.70)%],the autophagy(9.86±1.78)% and apoptosis (13.25±1.56)%of AGS cells were significantly enhanced after Hp treatment (t1=11.054,P1=0.000;t2=10.456,P2=0.000); after SN50 treatment, AGS autophagy(19.65±2.14)% and apoptosis(28.65±2.14)% were significantly attenuated (t1=3.403,P1=0.009;t2=2.799,P2=0.023). After combined treatment of Hp and SN50, AGS autophagy(20.65±2.14)% and apoptosis (29.65±2.14)%were significantly higher than those in the control group, but were lower than those in the Hp alone treatment group; the difference was significant (t1=5.164,P1=0.001;t2=5.890,P2=0.000;t3=4.718,P3=0.002;t4=5.738,P4=0.000). Conclusions Hp stimulation significantly promotes autophagy and apoptosis of AGS cells,the mechanism of which may be related to the activation of NF-κB pathway. |
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