生理学报
生理學報
생이학보
ACTA PHYSIOLOGICA SINICA
2001年
1期
37-40
,共4页
刘俊昌%张敏%郑新程%吴立玲%时安云%伍贻经%李玉明
劉俊昌%張敏%鄭新程%吳立玲%時安雲%伍貽經%李玉明
류준창%장민%정신정%오립령%시안운%오이경%리옥명
内毒素%血红素加氧酶%基因表达
內毒素%血紅素加氧酶%基因錶達
내독소%혈홍소가양매%기인표체
为了探讨在内毒素作用下的乳鼠心肌细胞(neonatal rat cardiomyocytes, NRCMs)血红素加氧酶-1 (heme oxygenase-1, HO-1)基因的表达及其在细胞损伤中的作用, 分别用 10、30及50 μg/ml的脂多糖(lipopolysaccharide, LPS), 10 μg/ml LPS +10 μmol/ml锌原卟啉Ⅸ(Zn-protoporphyrin-Ⅸ, ZnPPⅨ)和单纯10 μmol/ml ZnPPⅨ与培养的NRCMs共同孵育6 h, 以及10 μg/ml LPS与NRCMs共同孵育9 h和18 h。分别观察细胞HO-1 mRNA表达、MDA含量、LDH释放量与台盼蓝摄取率的变化。结果显示, 同样与细胞孵育6 h, LPS 10 μg/ml时HO-1 mRNA表达比对照组增加81.2%, 30 μg/ml时表达量增加126.3%, 50 μg/ml时表达量增加92.8%; LPS为10 μg/ml时, 孵育9 h后HO-1 mRNA的表达量比对照组增加93.6%, 孵育18 h后表达量增加105.8%。LPS 30、50 μg/ml, 10 μg/ml LPS+10 μmol/ml ZnPPⅨ与细胞孵育6 h及LPS 10 μg/ml孵育18 h后, 细胞MDA含量、LDH释放量与台盼蓝摄取率明显增加(P<0.01); 单纯10 μg/ml LPS与单纯10 μmol/ml ZnPPⅨ孵育6 h后, 上述指标均无明显升高。结果表明, LPS可诱导NRCMs HO-1 mRNA的表达, 且在较低LPS剂量范围内具有时间依赖性和浓度依赖性; NRCMs HO-1 mRNA的表达可减低LPS引起的细胞损伤, 这可能是细胞产生的一种自身保护性反应。
為瞭探討在內毒素作用下的乳鼠心肌細胞(neonatal rat cardiomyocytes, NRCMs)血紅素加氧酶-1 (heme oxygenase-1, HO-1)基因的錶達及其在細胞損傷中的作用, 分彆用 10、30及50 μg/ml的脂多糖(lipopolysaccharide, LPS), 10 μg/ml LPS +10 μmol/ml鋅原卟啉Ⅸ(Zn-protoporphyrin-Ⅸ, ZnPPⅨ)和單純10 μmol/ml ZnPPⅨ與培養的NRCMs共同孵育6 h, 以及10 μg/ml LPS與NRCMs共同孵育9 h和18 h。分彆觀察細胞HO-1 mRNA錶達、MDA含量、LDH釋放量與檯盼藍攝取率的變化。結果顯示, 同樣與細胞孵育6 h, LPS 10 μg/ml時HO-1 mRNA錶達比對照組增加81.2%, 30 μg/ml時錶達量增加126.3%, 50 μg/ml時錶達量增加92.8%; LPS為10 μg/ml時, 孵育9 h後HO-1 mRNA的錶達量比對照組增加93.6%, 孵育18 h後錶達量增加105.8%。LPS 30、50 μg/ml, 10 μg/ml LPS+10 μmol/ml ZnPPⅨ與細胞孵育6 h及LPS 10 μg/ml孵育18 h後, 細胞MDA含量、LDH釋放量與檯盼藍攝取率明顯增加(P<0.01); 單純10 μg/ml LPS與單純10 μmol/ml ZnPPⅨ孵育6 h後, 上述指標均無明顯升高。結果錶明, LPS可誘導NRCMs HO-1 mRNA的錶達, 且在較低LPS劑量範圍內具有時間依賴性和濃度依賴性; NRCMs HO-1 mRNA的錶達可減低LPS引起的細胞損傷, 這可能是細胞產生的一種自身保護性反應。
위료탐토재내독소작용하적유서심기세포(neonatal rat cardiomyocytes, NRCMs)혈홍소가양매-1 (heme oxygenase-1, HO-1)기인적표체급기재세포손상중적작용, 분별용 10、30급50 μg/ml적지다당(lipopolysaccharide, LPS), 10 μg/ml LPS +10 μmol/ml자원계람Ⅸ(Zn-protoporphyrin-Ⅸ, ZnPPⅨ)화단순10 μmol/ml ZnPPⅨ여배양적NRCMs공동부육6 h, 이급10 μg/ml LPS여NRCMs공동부육9 h화18 h。분별관찰세포HO-1 mRNA표체、MDA함량、LDH석방량여태반람섭취솔적변화。결과현시, 동양여세포부육6 h, LPS 10 μg/ml시HO-1 mRNA표체비대조조증가81.2%, 30 μg/ml시표체량증가126.3%, 50 μg/ml시표체량증가92.8%; LPS위10 μg/ml시, 부육9 h후HO-1 mRNA적표체량비대조조증가93.6%, 부육18 h후표체량증가105.8%。LPS 30、50 μg/ml, 10 μg/ml LPS+10 μmol/ml ZnPPⅨ여세포부육6 h급LPS 10 μg/ml부육18 h후, 세포MDA함량、LDH석방량여태반람섭취솔명현증가(P<0.01); 단순10 μg/ml LPS여단순10 μmol/ml ZnPPⅨ부육6 h후, 상술지표균무명현승고。결과표명, LPS가유도NRCMs HO-1 mRNA적표체, 차재교저LPS제량범위내구유시간의뢰성화농도의뢰성; NRCMs HO-1 mRNA적표체가감저LPS인기적세포손상, 저가능시세포산생적일충자신보호성반응。
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 μg/ml (H, 6 h) LPS and 10 μg/ml LPS +10 μmol/ml Zn-protoporphyrin-Ⅸ (ZnPPⅨ; L+I, 6 h) and 10 μmol/ml ZnPPⅨ alone (I, 6 h) were added to the medium for a 6-hour culture of NRCMs, and 10 μg/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 μg/ml. When the final concentrations of LPS were 10 and 30 μg/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 μg/ml, the HO-1 mRNA expression decreased to the level of 10 μg/ml group. When the final concentration was 10 μg/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 μg/ml LPS and 10 μg/ml LPS +10 μmol/ml ZnPPⅨ for 6 h and 10 μg/ml LPS for 18 h, the rate of Trypan blue stain uptake, MDA contents and LDH release significantly increased. With 10 μg/ml LPS alone and 10 μmol/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.