川芎嗪抑制CEACAM1表达保护脂多糖诱导的心肌细胞损伤

Ligustrazine protected cardiomyocytes from injury induced by lipopolysaccharide by down-regulating the expression of CEACAM1

目的探讨川芎嗪在脂多糖诱导的心肌细胞损伤中的作用及其机制。方法体外培养H9C2心肌细胞,以空白对照组(Sham)作为对照,予不同浓度(100~1 000 ng/m L)的脂多糖进行诱导,采用CCK8法检测心肌细胞损伤;给予不同浓度(20~80μmol/L)的川芎嗪进行干预,观察脂多糖刺激后心肌细胞损伤的变化;采用ELISA法检测培养液中白介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、转化生长因子-β1(TGF-β1)水平;荧光定量PCR检测心肌细胞中癌胚抗原相关细胞黏附分子1(CEACAM1)、磷酸化p38丝裂原活化蛋白激酶(p-p38 MAPK)、Caspase m RNA水平。结果与对照组相比,500 ng/m L的脂多糖导致心肌细胞存活率明显下降[(100.51±1.76)%vs(65.67±4.81)%,P<0.01],IL-1β[(25.17±1.77)pg/m L vs(60.50±3.67)pg/m L,P<0.01]和TNF-α[(34.33±2.48)pg/m L vs(64.67±2.49)pg/m L,P<0.01]的水平明显增加,TGF-β1水平下降[(69.00±3.07)pg/m L vs(39.50±1.87)pg/m L,P<0.01],心肌细胞中CEACAM1[(1.01±0.02)vs(1.30±0.03),P<0.01]、p-p38 MAPK[(1.01±0.02)vs(1.19±0.02),P<0.05]、Caspase3[(0.99±0.01)vs(1.25±0.03),P<0.05]的m RNA水平显著增加;给予40μmol/L的川芎嗪干预后,由脂多糖诱导的心肌细胞损伤程度减轻,细胞存活率增加[(65.67±4.80)%vs(81.17±2.42)%,P<0.05],IL-1β[(60.50±3.67)pg/m L vs(39.00±2.16)pg/m L,P<0.01]和TNF-α[(64.67±2.49)pg/m L vs(44.83±2.78)pg/m L,P<0.01]的水平降低,TGF-β1[(39.50±1.87)pg/m L vs(49.33±2.18)pg/m L,P<0.05]水平增加,CEACAM1[(1.30±0.03)vs(0.82±0.02),P<0.01]、p-p38 MAPK[(1.19±0.02)vs(0.98±0.03),P<0.05]、Caspase3[(1.25±0.04)vs(0.87±0.05),P<0.01]的m RNA水平显著减低。结论川芎嗪对脂多糖诱导的心肌细胞损伤有保护作用,该作用可能与脂多糖降低CEACAM1表达有关。

Objective To investigate the effect of Ligustrazine on cardiomyocytes injury induced by lipopolysaccharide（LPS） and its mechanism. Methods H9 C2 cardiomyocytes were cultured in vitro and induced by LPS（100～1 000 ng/m L）. Cardiomyocytes injury were detected by CCK8 kit. Ligustrazines（20～80 μmol/L） were added to survey its effect in cardiomyocytes injury. The levels of interleukin-1 β（IL-1 β）, tumor necrosis factor-α（TNF-α） and transforming growth factor-β1（TGF-β1） in the supernatant were measured by ELISA. The m RNA levels of CEACAM1, phosphorylated-p38 mitogen-activated protein kinases（p-p38 MAPK） and caspase3 in cardiomyocytes were measured by real time PCR. Results Compared with the control group（Sham）, 500 ng/m L LPS led to significantly deceased survive percentage of cardiomyocytes in a dose-dependent manner [（100.51±1.76）% vs（65.67±4.81）%, P〈0.01], significantly elevated levels of IL-1β [（25.17±1.77） pg/m L vs（60.50±3.67） pg/m L, P〈0.01] and TNF-α [（34.33±2.48） pg/m L vs（64.67±2.49） pg/m L, P〈0.01], significantly deceased level of TGF-β1[（69.00±3.07） pg/m L vs（39.50±1.87） pg/m L, P〈0.01], and significantly elevated m RNA levels of CEACAM1 [（1.01±0.02） vs（1.30±0.03）, P〈0.01], p-p38 MAPK [（1.01±0.02） vs（1.19±0.02）, P〈0.05] and caspase3 [（0.99±0.01） vs（1.25±0.03）, P〈0.05]. Treatment with 40 μmol/L Ligustrazines results in significantly increased survive percentage of cardiomyocytes [（65.67±4.80）% vs（81.17±2.42）%, P〈0.05], significantly decreased levels of IL-1β [（60.50±3.67） pg/m L vs（39.00±2.16） pg/m L, P〈0.01] and TNF-α [（64.67±2.49） pg/m L vs（44.83 ± 2.78） pg/m L, P〈0.01], significantly increased TGF-β1 level [（39.50 ± 1.87） pg/m L vs（49.33 ± 2.18） pg/m L, P〈0.05], as well as significantly decreased m RNA levels of CEACAM1 [（1.30 ± 0.03） vs（0.82 ± 0.02）, P〈0.01], p-p38 MAPK [（1.19±0.02） vs（0.98±0.03）, P〈0.05] and caspase3 [（1.25±0.04） vs（0.87±0.05）, P〈0.01]. Conclusion Ligustrazine can protect cardiomyocytes from injury induced by LPS, which might be related to its effect of down-regulating the expression of CEACAM1.