标题：川芎嗪抑制CEACAM1表达保护脂多糖诱导的心肌细胞损伤

      作者：刘峰 1，骆瑜 2，刘华东 1，高原 1，曹园芝 1，董少红 1，熊玮 1    (深圳市人民医院/暨南大学第二临床医学院心内科 1、老年病科 2，广东 深圳 518020)

      卷次： 2018年29卷17期

      【摘要】 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/mL)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(20~80 μmol/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 mRNA水平。结果 与对照组相比，500 ng/mL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/mL vs (60.50±3.67) pg/mL，P<0.01]和TNF-α [(34.33±2.48) pg/mL vs (64.67±2.49) pg/mL，P<0.01]的水平明显增加，TGF-β1水平下降[(69.00±3.07) pg/mL vs (39.50±1.87) pg/mL，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mRNA水平显著增加；给予40 μmol/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/mL vs (39.00±2.16) pg/mL，P<0.01]和TNF-α [(64.67±2.49) pg/mL vs (44.83±2.78) pg/mL，P<0.01]的水平降低，TGF-β1 [(39.50±1.87) pg/mL vs(49.33±2.18) pg/mL，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mRNA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。
      【关键词】 川芎嗪；心肌细胞；炎症；癌胚抗原相关细胞黏附分子1
      【中图分类号】 R542.2 【文献标识码】 A 【文章编号】 1003—6350（2018）17—2373—04

Ligustrazine protected cardiomyocytes from injury induced by lipopolysaccharide by down-regulating theexpression of CEACAM1.
LIU Feng 1, LUO Yu 2, LIU Hua-dong 1, GAO Yuan 1, CAO Yuan-zhi 1, DONG Shao-hong 1,XIONG Wei 1. Department of Cardiology 1, Department of Gerontology 2, Shenzhen People's Hospital, the Second AffiliatedHospital of Jinan Medical College, Shenzhen 518020, Guangdong, CHINA
【Abstract】 Objective To investigate the effect of Ligustrazine on cardiomyocytes injury induced by lipopoly-saccharide (LPS) and its mechanism. Methods H9C2 cardiomyocytes were cultured in vitro and induced by LPS (100~1 000 ng/mL). Cardiomyocytes injury were detected by CCK8 kit. Ligustrazines (20~80 μmol/L) were added to surveyits effect in cardiomyocytes injury. The levels of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and trans-forming growth factor-β1 (TGF-β1) in the supernatant were measured by ELISA. The mRNA levels of CEACAM1, phos-phorylated-p38 mitogen-activated protein kinases (p-p38 MAPK) and caspase3 in cardiomyocytes were measured by re-al time PCR. Results Compared with the control group (Sham), 500 ng/mL LPS led to significantly deceased survivepercentage of cardiomyocytes in a dose-dependent manner [(100.51±1.76)% vs (65.67±4.81)%, P<0.01], significantly el-evated levels of IL-1β [(25.17±1.77) pg/mL vs (60.50±3.67) pg/mL, P<0.01] and TNF-α [(34.33±2.48) pg/mL vs (64.67±2.49) pg/mL, P<0.01], significantly deceased level of TGF-β1 [(69.00±3.07) pg/mL vs (39.50±1.87) pg/mL, P<0.01], andsignificantly elevated mRNA 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 resultsin significantly increased survive percentage of cardiomyocytes [(65.67±4.80)% vs (81.17±2.42)%, P<0.05], significant-ly decreased levels of IL-1β [(60.50±3.67) pg/mL vs (39.00±2.16) pg/mL, P<0.01] and TNF-α [(64.67±2.49) pg/mL vs(44.83±2.78) pg/mL, P<0.01], significantly increased TGF-β 1 level [(39.50±1.87) pg/mL vs (49.33±2.18) pg/mL, P<0.05], as well as significantly decreased mRNA levels of CEACAM1 [(1.30 ± 0.03) vs (0.82 ± 0.02), P<0.01], p-p38MAPK [(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 Ligustra-zine can protect cardiomyocytes from injury induced by LPS, which might be related to its effect of down-regulating theexpression of CEACAM1.
      【Key words】 Ligustrazine; Cardiomyocyte; Inflammation; CEACAM1·论 著·基金项目：广东省深圳市科技创新课题(编号：JCYJ20160422151912249)