| 胡海英 胡旭堂 王志禄 谢宛霞 徐芳 蒙颖 朱海 白海渊 元朝波.S-亚硝基-N- 乙酰-DL- 青霉胺对RAW264.7 巨噬细胞亚型
分化的影响[J].,2014,14(31):6039-6043 |
| S-亚硝基-N- 乙酰-DL- 青霉胺对RAW264.7 巨噬细胞亚型
分化的影响 |
| Effect of S-nitroso-N-acetyl-DL-penicillamine (SNAP) on the SubtypeDifferentiation of RAW264.7 Macrophages |
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| DOI： |
| 中文关键词: 动脉粥样硬化 S-亚硝基-N-乙酰-DL-青霉胺 RAW264.7 巨噬细胞 M1 亚型 M2 亚型 |
| 英文关键词: Atherosclerosis(AS) SNAP RAW264.7 macrophage M1/M2 phenotypes |
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| 中文摘要: |
| 目的：探讨S- 亚硝基-N- 乙酰-DL- 青霉胺(SNAP)对巨噬细胞亚型分化的影响及其机制。方法：以RAW264.7 巨噬细胞为研
究对象，分为空白对照组、SNAP 组、SNAP+PBA(4- 苯基丁酸)组，采用不同浓度(30、100、300、400、500 umol/L)的SNAP 或300
umol/L SNAP+20 mmol/L PBA 对巨噬细胞进行干预24 h，应用RT-PCR法检测RAW264.7 巨噬细胞亚型分化标志物M1(iNOS，
CD86)、M2(Arg-I，MR)及CHOP mRNA 的表达，应用Western blot 技术检测iNOS 及ERS通路中相关蛋白CHOP、P-PERK 的表
达。结果：与空白对照组比较，SNAP组iNOS、CD86、CHOPmRNA的表达均明显降低(P＜0.05)，Arg-ImRNA表达明显升高(P<0.05)，而
MR mRNA表达升高，但差异无统计学意义(P>0.05)；与300 umol/L SNAP 组比较，300 umol/L+PBA组iNOS、CHOP mRNA 均无
明显变化(P>0.05)，CD86 mRNA升高，Arg-I、MR mRNA 均明显降低(P<0.05)。SNAP 组CHOP、iNOS、p-PERK蛋白表达均明显低
于对照组(P<0.05)，300 umol/L SNAP+20 mmol/L PBA 组与300 umol/L SNAP 组比较iNOS蛋白、p-PERK、CHOP蛋白表达升高
(P< 0.05)。结论：NO 可能通过内质网应激机制抑制巨噬细胞向M1 亚型分化。 |
| 英文摘要: |
| Objective:To investigate the effect and mechanism of SNAP on the subtype differentiation of RAW264.7
macrophages.Methods:RAW264.7 macrophages were plated in 12 wells plate as 106/ml for 24 h before intervention of SNAP in
different concentration (30, 100, 300, 400 and 500 umol/L) or 300 umol/L SNAP+20 mmol/L PBA. Total RNA of cells were extracted
after intervention for 24 hours. The mRNA expression of phonetype marker iNOS, CD86 (as M1 phenotypes markers), MR, and
Arginase-I (Arg-I) (as M2 phenotypes markers) were detected respectively by real time PCR. The protein expression of CHOP, p-PERK
were detected by western blotting.Results:Compared with the control group, the iNOS, CD86 and CHOP mRNA expression
significantly decreased(P<0.05), Arg-I mRNA expression increased, but no significant difference was found(P>0.05). Compared with 300
umol/L SNAP group, no remarkable difference was found in iNOS, CHOP mRNA expression of 300 umol/L+PBA group, but Arg-I, MR
mRNA expression both significantly decreased (P<0.05). The protein expression of CHOP, iNOS and p-PERK of SNAP group were all
significantly lower than those of the control group (P<0.05), compared with 300 umol/L SNAP group, the protein expression of CHOP,
p-PERK, iNOS increased.Conclusion:SNAP could suppress macrophage differentiation to M1 subtype through endoplasmic reticulum
stress (ERS). |
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