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颜佳博,邓奕辉,廖俞强,李钰佳,李定祥.基于RAGE/NF-κB/mTOR信号通路探讨滋阴活血解毒方对AGEs诱导VEC损伤的干预作用[J].湖南中医药大学学报英文版,2017,37(7):719-722.[Click to copy
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| 基于RAGE/NF-κB/mTOR信号通路探讨滋阴活血解毒方对AGEs诱导VEC损伤的干预作用 |
| 颜佳博,邓奕辉,廖俞强,李钰佳,李定祥 |
| (湖南中医药大学, 湖南 长沙 410208) |
| 摘要: |
| 目的 观察滋阴活血解毒方对糖基化终末产物（advanced glycosylation end products，AGEs）诱导人脐静脉内皮细胞（human umbilical vein endothelial cell，HUVEC）损伤的影响及RAGE/NF-κB/mTOR信号通路的变化，探讨该方对AGEs诱导静脉内皮细胞（vein endothelial cell，VEC）损伤的保护作用。方法 培养的HUVEC细胞以100 mg/ml的AGEs诱导损伤，随机分为7个组，除空白组外，其中3组分别加入RAGE抑制剂FPS-ZM1、NF-κB抑制剂PDTC和mTOR抑制剂雷帕霉素（RAP），余下3组加入各种抑制剂后加入50 mg/mL滋阴活血解毒方浓缩药液。以western-blot法检测RAGE和mTOR的表达，以免疫组化方法检测NF-κB核转移状况。结果 细胞添加AGEs后细胞内出现异常增殖，细胞数量明显增多，多数细胞变形皱缩，胞膜轮廓模糊，细胞内出现粗糙样颗粒变化；抑制剂组及抑制剂加中药组都表现出不同程度的逆转这种损伤的作用，特别是抑制剂加中药组，见梭形正常细胞较多，圆缩形细胞减少。中药加抑制剂组RAGE表达量比单纯抑制剂组显著降低（P<0.01）；中药加抑制剂组mTOR表达量比单纯抑制剂组显著降低（P<0.01）；中药加抑制剂NF-κB的核转移阳性检测率比单纯抑制剂组显著降低（P<0.01）。结论 滋阴活血解毒方能降低AGEs对VEC的损伤，降低RAGE、NF-κ、mTOR的表达。 |
| 关键词: 滋阴活血解毒方 AGEs 内皮细胞损伤 RAGE mTOR NF-κB |
| DOI：10.3969/j.issn.1674-070X.2017.07.006 |
| Received:February 17, 2017 |
| 基金项目:湖南省教育厅项目（16K063）；湖南省中医药科研基金（201606）；中西医结合基础湖南省重点学科资助。 |
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| Effect of Ziyin Huoxue Jiedu Decoction on VEC Injury Induced by AGEs Based onRAGE/NF-κB/mTOR Signaling Pathway |
| YAN Jiabo,DENG Yihui,LIAO Yuqiang,LI Yujia,LI Dingxiang |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To observe the effect of Ziyin Jiedu decoction on the damage of human umbilical vein endothelial cell (HUVEC) induced by AGEs and the change of RAGE/NF-κB/mTOR pathway, and to explore the protective effect of this prescription on AGEs-induced VEC injury. Methods The damage of HUVEC cells were induced by 100 mg/mL AGEs and randomly divided into seven groups. Three of these groups excepted blank group were added RAGE inhibitors FPS-ZM1, NF-κB inhibitor PDTC and mTOR inhibitor rapamycin (RAP) respectively, and the remaining three groups were treated with RAGE inhibitor FPS-ZM1, NF-κB inhibitor PDTC and mTOR inhibitor rapamycin (RAP) then added 50mg/mL concentrated Ziyin Huoxue Jiedu decoction to test. The expression of RAGE and mTOR was detected by Western-blot, and NF-κB nuclear translocation was detected by immunohistochemistry. Results The cells added with AGEs show abnormal growth, such as metamorphosis and shrink cells, membrane with hazy outline, roughness particles, which increase obviously. with increased cells. Inhibitor group and inhibitor added with Chinese medicine group could reverse this lesion with different degree. Inhibitor added with Chinese medicine group showed manny fusiform normal cells and few round and shrink cells. The expression of RAGE in inhibitor added with Chinese medicine group showed an obvious increase than inhibitor group (P<0.01). The expression of mTOR in inhibitor added with Chinese medicine group showed a significant increase than inhibitor group (P<0.01). The positive detection rate of NF-κB drug-containing inhibitor was significantly lower than that of inhibitor group (P<0.01). Conclusion Ziyin Huoxue Jiedu decoction could reduce VEC damage induced by AGEs, and also could decrease the expression of RAGE, NF-κB, mTOR. |
| Key words: Ziyin Huoxue Jiedu decoction AGEs endothelial cell injury RAGE mTOR NF-κB |
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