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[脂多糖诱导新生大鼠心肌细胞血红素加氧酶-1的表达]

[Expression of heme oxygenase-1 in neonatal rat cardiocytes induced by lipopolysaccharide].

作者信息

Li Y M, Liu J C, Zhang M, Zheng X C, Wu L L, Shi A Y, Wu Y J

机构信息

Department of Pathophysiology, Beijing Medical University, Beijing 100083.

出版信息

Sheng Li Xue Bao. 2001 Feb;53(1):37-40.

Abstract

To study the alterations of heme oxygenase-1 mRNA in neonatal rat cardiomyocytes (NRCMs) induced by lipopolysaccharide (LPS) and the role of heme oxygenase-1 (HO-1) in the LPS induced disorders of myocardium function, 10 (L, 6 h), 30 (M, 6 h), 50 micrograms/ml (H, 6 h) LPS and 10 micrograms/ml LPS + 10 mumol/ml Zn-protoporphyrin-IX (ZnPPIX; L + I, 6 h) and 10 mumol/ml ZnPPIX alone (I, 6 h) were added to the medium for a 6-hour culture of NRCMs, and 10 micrograms/ml LPS for 9 h (L, 9 h) and 18 h (L, 18 h) cultures. LDH release and MDA contents of the cells were measured. When NRCMs were collected, Trypan blue stain method was used to examine the mortality (the rate of Trypan blue uptake) of NRCMs. HO-1 mRNA expression was examined by Northern blot. The results showed that HO-1 mRNA expression of NRCMs increased gradually along with the increase of LPS concentration below the level of 30 micrograms/ml. When the final concentrations of LPS were 10 and 30 micrograms/ml, the HO-1 mRNA expression of NRCMs increased by 81.2% and 126.3% respectively compared with control. When the final concentration of LPS was 50 micrograms/ml, the HO-1 mRNA expression decreased to the level of 10 micrograms/ml group. When the final concentration was 10 micrograms/ml, the HO-1 mRNA expression increased gradually along with the culture time. After a 9- or 18-hour culture, the HO-1 mRNA expression of NRCMs increased by 93.6% and 105.8% respectively compared with control. Only when NRCMs had been cultured with 30, 50 micrograms/ml LPS and 10 micrograms/ml LPS + 10 mumol/ml ZnPPIX for 6 h and 10 micrograms/ml LPS for 18 h, the rate of Trypan blue stain uptake, MDA contents and LDH release significantly increased. With 10 micrograms/ml LPS alone and 10 mumol/ml ZnPPIX alone for 6 h, the above parameters were not significantly increased (P > 0.05). The results demonstrate that LPS induces HO-1 mRNA expression of NRCMs dose- and time-dependently to some extent. The inducible HO can protect NRCMs from injury and thus play an important role in pathogenesis of myocardium under LPS.

摘要

为研究脂多糖(LPS)诱导新生大鼠心肌细胞(NRCMs)血红素加氧酶-1 mRNA的变化以及血红素加氧酶-1(HO-1)在LPS诱导的心肌功能紊乱中的作用,将10(L,6小时)、30(M,6小时)、50微克/毫升(H,6小时)LPS以及10微克/毫升LPS + 10微摩尔/毫升锌原卟啉-IX(ZnPPIX;L + I,6小时)和单独的10微摩尔/毫升ZnPPIX(I,6小时)加入培养基中对NRCMs进行6小时培养,以及10微克/毫升LPS分别进行9小时(L,9小时)和18小时(L,18小时)培养。检测细胞的乳酸脱氢酶(LDH)释放量和丙二醛(MDA)含量。收集NRCMs时,采用台盼蓝染色法检测NRCMs的死亡率(台盼蓝摄取率)。通过Northern印迹法检测HO-1 mRNA表达。结果显示,在LPS浓度低于30微克/毫升时,NRCMs的HO-1 mRNA表达随LPS浓度增加而逐渐升高。当LPS终浓度为10和30微克/毫升时,NRCMs的HO-1 mRNA表达分别比对照增加81.2%和126.3%。当LPS终浓度为50微克/毫升时,HO-1 mRNA表达降至10微克/毫升组水平。当终浓度为10微克/毫升时,HO-1 mRNA表达随培养时间逐渐升高。培养9小时和18小时后,NRCMs的HO-1 mRNA表达分别比对照增加93.6%和105.8%。仅当NRCMs用30、50微克/毫升LPS以及10微克/毫升LPS + 10微摩尔/毫升ZnPPIX培养6小时和10微克/毫升LPS培养18小时时,台盼蓝染色摄取率、MDA含量和LDH释放量显著增加。单独用10微克/毫升LPS和单独用10微摩尔/毫升ZnPPIX培养6小时,上述参数无显著增加(P > 0.05)。结果表明,LPS在一定程度上呈剂量和时间依赖性诱导NRCMs的HO-1 mRNA表达。诱导型HO可保护NRCMs免受损伤,因此在LPS作用下的心肌发病机制中起重要作用。

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