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纸质出版日期:2017
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南娜, 赵萍, 张睿. 银杏叶提取物对NF-B活性抑制人视网膜色素上皮细胞炎症反应的影响[J]. 中国实验方剂学杂志, 2017,23(18):163-167.
NAN Na, ZHAO Ping, ZHANG Rui. Inflammation Inhibitory Effect of Ginkgo Leaves Extract on NF-B Activity in Human Retinal Pigment Epithelial Cells[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(18): 163-167.
南娜, 赵萍, 张睿. 银杏叶提取物对NF-B活性抑制人视网膜色素上皮细胞炎症反应的影响[J]. 中国实验方剂学杂志, 2017,23(18):163-167. DOI: 10.13422/j.cnki.syfjx.2017180163.
NAN Na, ZHAO Ping, ZHANG Rui. Inflammation Inhibitory Effect of Ginkgo Leaves Extract on NF-B Activity in Human Retinal Pigment Epithelial Cells[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(18): 163-167. DOI: 10.13422/j.cnki.syfjx.2017180163.
目的:观察银杏叶提取物对高糖诱导的人视网膜色素上皮细胞ARPE-19炎症的影响及核转录因子-kappaB(NF-κB)通路的变化,探讨其作用机制。方法:体外培养ARPE-19细胞株,分为正常组(5.5 mmol · L-1葡萄糖),高糖组(30 mmol ·L-1葡萄糖),银杏叶提取物不同剂量组组(30 mmol · L-1葡萄糖+12.5,25,50,100 mg · L-1银杏叶提取物)。噻唑蓝(MTT)比色法检测细胞活性;流式细胞术测定细胞中肿瘤坏死因子-α(TNF-α),白细胞介素-1β(IL-1β)的含量;实时荧光定量聚合酶链式反应(Real-time PCR)和蛋白质免疫印迹法(Western blot)测定炎症因子(NIK/p-NIK,IKKα/p-IKKα,IκB/p-IκB)表达;Western blot检测NF-κB磷酸化及核NF-κB表达。结果:与正常组比较,高糖组吸光度A明显降低(P<0.05),细胞TNF-α,IL-1β含量明显升高(P<0.05),p-NIK,p-IKKα,p-IκB表达水平明显升高(P<0.05),p-NF-κB,核NF-κB表达水平明显升高(P<0.05);与高糖组比较,12.5,25,50,100 mg ·L-1银杏叶提取物组吸光度A明显升高(P<0.05);细胞TNF-α,IL-1β含量明显降低(P<0.05),p-NIK,p-IKKα,p-IκB水平明显降低(P<0.05),p-NF-κB,核NF-κB表达水平明显降低(P<0.05)。结论:银杏叶提取物能通过抑制NF-κB活性抑制炎症反应,保护视网膜色素上皮细胞,用于糖尿病视网膜病变的防治。
Objective: To observe the effect of ginkgo leaves extract on the inflammation of human esophageal epithelial cell ARPE-19 line and the changes in nuclear factor-kappaB(NF-κB)pathway
in order to explore its mechanism.Method: In vitro
ARPE-19 cells was divided into normal control group (5.5 mmol · L-1 glucose)
high-glucose group (30 mmol · L-1 glucose) and ginkgo leaves extract group (30 mmol · L-1 glucose+12.5
25
50
100 mg · L-1 ginkgo leaves extract). MTT assay was used to detect cell viability. The levels of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in the cells were determined by flow cytometry. NIK/p-NIK
IKKα/p-IKKα and IκB/p-IκB expressions were measured by Real-time PCR and Western blot. Western blot was used to detect the phosphorylation of NF-κB and the expression of nuclear NF-κB.Result: The Avalue of the high-glucose group was significantly lower than that of the control group
ginkgo leaves extract groups (12.5
25
50
100 mg · L-1) were significantly higher than those of the high-glucose group (P<0.05). Levels of TNF-α and IL-1β of the high-glucose group was significantly higher than that of the control group
andthose of ginkgo leaves extract group (12.5
25
50
100 mg · L-1) were significantly lower than those of high glucose group (P<0.05). Compared with the control group
the expressions of p-NIK
p-IKKα
p-IκB in the high-glucose group increased
and the expressions of p-NIK
p-IKKα
p-IκB were decreased in ginkgo leaves extract(12.5
25
50 and 100 mg · L-1)groups (P<0.05). The phosphorylation and nuclear NF-κB protein expression were increased in the high-glucose group compared with the control group
and the NF-κB expression were decreased in ginkgo leaves extract groups (12.5
25
50
100 mg · L-1
P<0.05).Conclusion: ginkgo leaves extract can inhibit the inflammatory response by inhibiting NF-κB activity and protect retinal pigment epithelial cells
so as to prevent and treat diabetic retinopathy.
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