硫化氢抑制氧化型低密度脂蛋白诱导的人巨噬细胞单核细胞趋化蛋白-1分泌的作用

Effect of Hydrogen Sulfide Inhibiting Monocyte Chemoattractant Protein-1 Secretion in Human Macrophages Stimulated by Oxidized Low Density Lipoprotein

目的研究气体信号分子硫化氢(H2S)对氧化型低密度脂蛋白(ox-LDL)诱导的人单核细胞白血病细胞(THP-1)来源的人巨噬细胞单核细胞趋化蛋白-1(MCP-1)分泌的影响。方法将THP-1来源的人巨噬细胞分为4组:正常对照组、ox-LDL组、ox-LDL+H2S 100组及ox-LDL+H2S 500组。正常对照组:细胞于基础培养基中培养48 h;ox-LDL组:在基础培养基中加入50 mg.L-1ox-LDL,培养48 h;ox-LDL+H2S 100组在基础培养基中先加入100μmol.L-1NaHS,孵育30 min后再加入50 mg.L-1ox-LDL,培养48 h;ox-LDL+H2S 500组在基础培养基中先加入500μmol.L-1NaHS,孵育30 min后再加入50 mg.L-1ox-LDL,培养48 h。用ELISA法检测细胞上清液中MCP-1水平。结果与正常对照组[(34.58±6.77)μmol.L-1]比较,ox-LDL组[(66.27±7.29)μmol.L-1]、ox-LDL+H2S 100组[(49.45±3.08)μmol.L-1]及ox-LDL+H2S 500组[(46.64±5.47)μmol.L-1]细胞上清液中MCP-1水平均明显升高(Pa<0.05);与ox-LDL组比较,ox-LDL+H2S 100组及ox-LDL+H2S 500组细胞上清液中MCP-1水平均明显降低(Pa<0.05)。ox-LDL+H2S 100组细胞上清液中MCP-1水平与ox-LDL+H2S 500组比较差异无统计学意义(P>0.05)。结论ox-LDL可诱导人巨噬细胞MCP-1分泌增加;给予外源性H2S的供体NaHS可明显抑制ox-LDL诱导的MCP-1分泌增加。

Objective To explore the effect of hydrogen sulfide （ H2 S） on oxidized - low density lipoprotein （ ox - LDL） - stimulated monocyte chemoattractant protein - 1 （ MCP - 1 ） secretion in THP - 1 - derived macrophage. Methods THP - 1 - derived macrophages were divided into 4 groups：normal control group, ox - LDL group, ox - LDL ＋ H2 S 100 group and ox - LDL ＋ H2 S 500 group. In the normal control group,cells were cultured in the basal medium for 48 hours;in the ox- LDL group,cells were treated with ox -LDL （50 mg· L-1） for 48 hours;in the ox -LDL ＋ H2S 100 group,cells were pretreated with Naris （100 μmol ·L^-1 ） for 30 minutes and then added ox - LDL （50 mg ·L ^- 1 ） to culture for 48 hours ; in the ox - LDL ＋ H2 S 500 group, cells were pretreated with NariS （500 μmol · L^-1 ） for 30 minutes and then added ox - LDL （50 mg· L^-1 ） to culture for 48 hours. The content of MCP - 1 in cell supcrnatant was examined by enzyme linked im- munosorbent assay. Results Compared with the control group [ （ 34.58 ± 6.77 ） μ mol· L^-1 ], MCP - 1 content increased greatly in the ox - LDL group [ （66.27 ＋ 7.29 ） μ mol ·L^-1 ], the ox - LDL ＋ H2 S 100 group [ （49.45 ±3.08 ） μ mol · L^- 1 ] and the ox - LDL ＋ H2 S 500 group [ （46.64 ± 5.47 ） μ mol ·L ^- 1 ] （ Pa 〈 0.05 ） ; compared with the ox - LDL group, MCP - 1 content significantly decreased in the ox - LDL ＋ H2S 100 group and the ox - LDL ＋ H2S 500 group （ P 〈 0.05 ）. There was no significant difference of MCP - 1 content between the ox - LDL ＋ H2S 100 group and the ox - LDL ＋ H2 S 500 group （ P 〉 0.05 ）. Conclusions ox - LDL promoted the secretion of MCP - 1 in macrophages,while the addition of Naris suppressed the increase of MCP- 1 secretion stimulated by ox- LDL.