黄连素对间歇性缺氧诱导的单核细胞分化和环氧合酶2表达的影响

摘要

目的 探讨黄连素对间歇性缺氧诱导的单核细胞THP1分化和环氧合酶2表达的影响.方法 利用三气细胞培养箱在间歇性缺氧条件下培养单核细胞系THP1细胞0、12、24、48 h,以正常氧环境培养作为对照.采用流式细胞技术检测单核细胞分化的标志蛋白CD14的表达变化,采用蛋白质印迹法检测环氧合酶2蛋白的表达变化.利用黄连素同时处理间歇性缺氧或正常氧状态培养的THP1细胞,采用流式细胞技术和蛋白质印迹法检测黄连素对间歇性缺氧诱导的单核细胞THP1分化和环氧合酶2表达的影响.结果 间歇性缺氧24、48 h的CD14阳性细胞比例明显高于间歇性缺氧0 h[(16.06±0.34)％、(15.75±2.47)％比(3.26±0.30)％],差异均有统计学意义(均P＜0.05),间歇性缺氧12 h的CD14阳性细胞比例[(3.80±0.47)％]与间歇性缺氧0h比较,差异无统计学意义(P＞0.05).间歇性缺氧24、48 h后THP1细胞环氧合酶2蛋白相对表达量明显高于间歇性缺氧0 h[(190±22)％、(295±23)％比(100±8)％],差异均有统计学意义(均P ＜0.05),间歇性缺氧12 h的THP1细胞环氧合酶2蛋白相对表达量[(129±11)％]与间歇性缺氧0h比较,差异无统计学意义(P＞0.05).正常氧状态培养48 h与正常氧状态+黄连素(3 μmol/L)培养48 h的CD14阳性细胞比例比较[(3.57±0.29)％比(3.31±0.22)％],差异无统计学意义(P＞0.05).间歇性缺氧48 h后CD14阳性细胞比例[(15.30±1.47)％]明显高于正常氧状态培养48 h,间歇性缺氧+黄连素(3μmol/L)培养48 h的CD14阳性细胞比例[(3.68±0.35)％]明显低于间歇性缺氧48 h,差异均有统计学意义(均P＜0.05).正常氧状态培养48 h与正常氧状态+黄连素(3 μmol/L)培养48 h的环氧合酶2蛋白相对表达量比较[(100±8)％比(104±17)％],差异无统计学意义(P＞0.05).间歇性缺氧48 h后环氧合酶2蛋白相对表达量[(162±4)％]明显高于正常氧状态培养48 h,间歇性缺氧+黄连素(3μmol/L)培养48 h的环氧合酶2蛋白相对表达量[(114±14)％]明显低于间歇性缺氧48 h,差异均有统计学意义(均P＜0.05).结论 间歇性缺氧能促进THP1分化及环氧合酶2表达,黄连素能有效抑制间歇性缺氧对THP1分化及环氧合酶2表达的促进作用.%Objective To investigate the influence of berberine on monocyte THP1 differentiation and cyclooxygenase-2(COX-2) expression induced by intermittent hypoxia.Methods THP1 cells were cultured in intermittent hypoxia condition(observation group) and normal oxygen condition(control group).After 0,12,24,48 h culture,expression of cell differentiation protein biomarker CD14 was tested by flow cytometry;expression of COX-2 protein was tested by western blotting.The cells were cultured with berberine.Changes of CD14 and COX-2 expression were analyzed by flow cytometry and western blotting.Results CD14 positive ratio in intermittent hypoxia 24,48 h group was significantly higher than that in 0 h group [(16.06 ± 0.34) ％,(15.75 ± 2.47) ％ vs (3.26 ± 0.30)％] (P ＜ 0.05).CD14 positive ratio in intermittent hypoxia 12 h group [(3.80 ± 0.47)％] showed no significant difference compared to that in 0 h group (P ＞ 0.05).COX-2 expression level in intermittent hypoxia 24,48 h group was significantly higher than that in 0 h group [(190 ± 22) ％,(295 ± 23) ％ vs (100 ± 8) ％] (P ＜0.05).COX-2 expression level in intermittent hypoxia 12 h group[(129 ±11)％] showed no significant difference compared to that in 0 h group(P ＞ 0.05).CD14 positive ratio showed no significant difference between normal oxygen 48 h group and normal oxygen + berberine(3 μmol/L) 48 h group[(3.57 ±0.29)％ vs (3.31 ±0.22)％] (P ＞ 0.05).CD14 positive ratio in intermittent hypoxia 48 h group [(15.30 ± 1.47)％] was significantly higher than that in normal oxygen 48 h group(P ＜ 0.05).CD14 positive ratio in intermittent hypoxia + berberine (3 μmol/L) 48 h group [(3.68 ± 0.35) ％] was significantly lower than that in intermittent hypoxia 48 h group(P ＜ 0.05).COX-2 expression level showed no significant difference between normal oxygen 48 h group and normal oxygen + berberine (3 μmol/L) 48 h group[(100 ± 8)％ vs (104 ± 17)％] (P ＞ 0.05).COX-2 expression level in intermittent hypoxia 48 h group[(162 ±4)％] was significantly higher than that in normal oxygen 48 h group(P ＜ 0.05).COX-2 expression level in intermittent hypoxia + berberine (3 μmol/L) 48 h group[(114 ± 14) ％] was significantly lower than that in intermittent hypoxia 48 h group (P ＜ 0.05).Conclusion Intermittent hypoxia promotes THP1 cell differentiation and COX-2 expression,which can be inhibited bv berberine.