中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2011年
2期
107-110
,共4页
何榕%毛节明%王广%高炜
何榕%毛節明%王廣%高煒
하용%모절명%왕엄%고위
晚期糖基化终产物%单核细胞趋化蛋白1%白细胞介素8%血管平滑肌细胞
晚期糖基化終產物%單覈細胞趨化蛋白1%白細胞介素8%血管平滑肌細胞
만기당기화종산물%단핵세포추화단백1%백세포개소8%혈관평활기세포
Advanced glycation end products%Monocyte chemoattractant protein 1%Interleukin-8%Vascular smooth muscle cells
目的 观察晚期糖基化终产物(AGE)对血管平滑肌细胞(VSMC)分泌单核细胞趋化蛋白1(MCP-1)和白细胞介素8(IL-8)的影响并初步探讨可能的细胞内信号转导机制.方法 分离、培养原代大鼠VSMC并进行鉴定.采用糖基化血清白蛋白(GSA)模拟AGE,观察不同浓度GSA(10、100和500 μg/ml)对VSMC分泌MCP-1和IL-8的影响和时间曲线;并对其进行细胞增殖率校正以消除VSMC数量增加对测定的影响;p38MAPK抑制剂(SB203580)、ERK1/2抑制剂(PD98059)及NF-κB抑制剂(PDTC)、Proteasome抑制剂(MG132)预处理后观察GSA刺激VSMC分泌MCP-1和IL-8水平的改变.结果 与空白对照组相比,100 μg/ml GSA作用24 h VSMC分泌MCP-1(13.01ng/ml±0.12ng/ml比7.02 ng/ml±0.26 ng/ml,P＜0.05)和IL-8(12.6ng/ml±0.86 ng/ml比3.07 ng/ml±0.35ng/ml,P＜0.05)水平最高.经过细胞增殖校正后,GSA仍然能够促进VSMC表达MCP-1和IL-8.MAPK抑制剂和NF-κB抑制剂预处理后发现PDTC(10 μmol/L)、SB203580(5 μmol/L)以及MG132(10 μmol/L)可以抑制GSA刺激VSMC表达MCP-1和IL-8.结论 GSA可以促进VSMC分泌致炎性趋化因子MCP-1和IL-8,这种作用独立于细胞增殖,可能是通过激活细胞内p38MAPK信号转导通路,促进核因子NF-κB的活化而实现.
目的 觀察晚期糖基化終產物(AGE)對血管平滑肌細胞(VSMC)分泌單覈細胞趨化蛋白1(MCP-1)和白細胞介素8(IL-8)的影響併初步探討可能的細胞內信號轉導機製.方法 分離、培養原代大鼠VSMC併進行鑒定.採用糖基化血清白蛋白(GSA)模擬AGE,觀察不同濃度GSA(10、100和500 μg/ml)對VSMC分泌MCP-1和IL-8的影響和時間麯線;併對其進行細胞增殖率校正以消除VSMC數量增加對測定的影響;p38MAPK抑製劑(SB203580)、ERK1/2抑製劑(PD98059)及NF-κB抑製劑(PDTC)、Proteasome抑製劑(MG132)預處理後觀察GSA刺激VSMC分泌MCP-1和IL-8水平的改變.結果 與空白對照組相比,100 μg/ml GSA作用24 h VSMC分泌MCP-1(13.01ng/ml±0.12ng/ml比7.02 ng/ml±0.26 ng/ml,P＜0.05)和IL-8(12.6ng/ml±0.86 ng/ml比3.07 ng/ml±0.35ng/ml,P＜0.05)水平最高.經過細胞增殖校正後,GSA仍然能夠促進VSMC錶達MCP-1和IL-8.MAPK抑製劑和NF-κB抑製劑預處理後髮現PDTC(10 μmol/L)、SB203580(5 μmol/L)以及MG132(10 μmol/L)可以抑製GSA刺激VSMC錶達MCP-1和IL-8.結論 GSA可以促進VSMC分泌緻炎性趨化因子MCP-1和IL-8,這種作用獨立于細胞增殖,可能是通過激活細胞內p38MAPK信號轉導通路,促進覈因子NF-κB的活化而實現.
목적 관찰만기당기화종산물(AGE)대혈관평활기세포(VSMC)분비단핵세포추화단백1(MCP-1)화백세포개소8(IL-8)적영향병초보탐토가능적세포내신호전도궤제.방법 분리、배양원대대서VSMC병진행감정.채용당기화혈청백단백(GSA)모의AGE,관찰불동농도GSA(10、100화500 μg/ml)대VSMC분비MCP-1화IL-8적영향화시간곡선;병대기진행세포증식솔교정이소제VSMC수량증가대측정적영향;p38MAPK억제제(SB203580)、ERK1/2억제제(PD98059)급NF-κB억제제(PDTC)、Proteasome억제제(MG132)예처리후관찰GSA자격VSMC분비MCP-1화IL-8수평적개변.결과 여공백대조조상비,100 μg/ml GSA작용24 h VSMC분비MCP-1(13.01ng/ml±0.12ng/ml비7.02 ng/ml±0.26 ng/ml,P＜0.05)화IL-8(12.6ng/ml±0.86 ng/ml비3.07 ng/ml±0.35ng/ml,P＜0.05)수평최고.경과세포증식교정후,GSA잉연능구촉진VSMC표체MCP-1화IL-8.MAPK억제제화NF-κB억제제예처리후발현PDTC(10 μmol/L)、SB203580(5 μmol/L)이급MG132(10 μmol/L)가이억제GSA자격VSMC표체MCP-1화IL-8.결론 GSA가이촉진VSMC분비치염성추화인자MCP-1화IL-8,저충작용독립우세포증식,가능시통과격활세포내p38MAPK신호전도통로,촉진핵인자NF-κB적활화이실현.
Objective To investigate the effects of advanced glycation end products (AGEs) on the secretion of monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8) in vascular smooth muscle cells (VSMCs) and explore its possible intracellular signaling mechanism. Methods Primary rat VSMCs were isolated and identified. VSMCs were treated with glycation serum albumin (GSA), an important component of AGEs, in series of concentrations and time. The role of MAPK and NF-κB inhibitors was confirmed. The levels of MCP-1 and IL-8 were determined by enzyme-linked immunosorbent assay (ELISA). Results VSMCs were treated with GSA at the doses of 10 μg/ml, 100 μg/ml and 500 μg/ml respectively. In comparison with the control group, the levels of MCP-1 ( 13.01 ng/ml ± 0.12 ng/ml vs 7. 02 ng/ml ±0. 26 ng/ml, P＜0.05) and IL-8 (12. 6 ng/ml ±0. 86 ng/ml vs 3. 07 ng/ml ±0.35 ng/ml,P＜0.05) increased in the GSA-treated group, especially at the concentration of 100 μg/ml. After adjustment for cells proliferation, the levels of MCP-1 and IL-8 were still higher in the GSA-treated group.After a pretreatment of PDTC ( 10 μmol/L), SB203580 (5 μmol/L) and MG132 ( 10 μmol/L), the levels of MCP-1 and IL-8 decreased. However, it had no change when pretreated with PD98059 (20 μmol/L).Conclusion GSA promotes the secretion of MCP-1 and IL-8 in VSMCs. Such an effect is not dependent on cellular proliferation. It may be realized through an activation of NF-κB by p38MAPK-sensitive intracellular signaling pathway.
