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中国临床药理学与治疗学 ›› 2017, Vol. 22 ›› Issue (12): 1346-1351.

• 基础研究 • 上一篇    下一篇

右美托咪定对A549细胞缺氧/复氧损伤时JNK的影响

罗梓垠1,2,项冰倩1,高 慧1,邱晓晓1,钱小英3,王万铁1   

  1. 1 温州医科大学缺血-再灌注损伤研究所,温州 325035,浙江;2 湖北省中西医结合医院病理科,武汉 430015,湖北;3 温州市人民医院呼吸内科,温州 325000 ,浙江
  • 收稿日期:2017-04-06 修回日期:2017-05-11 出版日期:2017-12-26 发布日期:2018-01-02
  • 通讯作者: 王万铁,男,教授,主任医师,博导,研究方向:脏器缺血/再灌注损伤的临床与基础。 Tel:0577-86689817 E-mail:wwt@wmu.edu.cn 钱小英,女,主任医师,研究方向:呼吸疾病的临床与基础。 Tel:13858841637
  • 作者简介:罗梓垠,女,硕士,研究方向:缺血/再灌注损伤。 E-mail:385115094@qq.com
  • 基金资助:

    浙江省公益技术应用研究项目(2013C33168);浙江省新苗人才计划项目(2014R413043);温州市公益性科技计划项目(Y20140652)

Effects of dexmedetomidine on JNK in A549 cells with hypoxia/reoxygenation injury

LUO Ziyin 1,2,  XIANG Bingqian1, GAO Hui1, QIU Xiaoxiao1, QIAN Xiaoying3, WANG Wantie1   

  1. 1 Ischemia/Reperfusion Injury Research Institute of Wenzhou Medical University, Wenzhou 325035, Zhejiang, China; 2 Pathology Department of Hubei Provincial Hospital of Integrated Chinese & Western Medicine, Wuhan 430015, Hubei, China; 3 Respiratory Department of Whenzhou People's Hospital, Wenzhou 325000, Zhejiang, China
  • Received:2017-04-06 Revised:2017-05-11 Online:2017-12-26 Published:2018-01-02

摘要:

目的: 观察右美托咪定(dexmedetomidine,DEX)对缺氧/复氧所致的A549细胞损伤的干预作用以及对c-Jun氨基末端激酶(JNK)的影响。方法:培养A549细胞株,将细胞随机分为4组(n=10):正常对照组(N组),DEX组(D组),缺氧/复氧损伤组(H组),缺氧/复氧损伤+DEX干预组(HD组)。造模结束后,在倒置显微镜下观察细胞形态学的变化。CCK-8法检测A549细胞活力。原位末端标记(TUNEL)法检测A549细胞的凋亡指数(AI)。蛋白免疫印迹法(Western blot)检测细胞内葡萄糖调节蛋白78(GRP78)、p-JNK、caspase-3蛋白的表达水平,逆转录-聚合酶链反应(RT-PCR)检测GRP78、JNK mRNA的表达水平。结果:与N组比较,H组贴壁细胞数量明显减少,细胞形态发生改变。A549细胞的吸光度(OD)值明显下降(P<0.01),AI值升高(P<0.01),凋亡细胞数明显增加(P<0.01)。HD组与H组相比,细胞损伤减轻,OD值上调(P<0.01),凋亡细胞数相对减少,AI值下调(P<0.01)。p-JNK、caspase-3蛋白和JNK mRNA表达下降(P<0.01)。结论:DEX可有效减轻A549细胞缺氧/复氧损伤,机制可能与其抑制JNK通路激活所致的细胞凋亡有关。

关键词: JNK, 右美托咪定, 缺氧/复氧损伤, A549细胞, 细胞凋亡

Abstract:

AIM: To investigate the effects and c-Jun N-terminal kinase(JNK)  expression of dexmedetomidine (DEX) on A549 cells with hypoxia/reoxygenation(H/R) injury. METHODS: A549 cells were cultivated and were randomly divided into four groups (n=10): control group (N), DEX group (D), hypoxia/reoxygenation injury group (H), hypoxia/reoxygenation injury+DEX interfere group (HD). After all models were completed, the morphological changes of A549 cells were observed under the inverted microscope. Cell activity was detected by CCK-8 and the apoptosis index (AI) was detected by in situ end labeling (TUNEL) method. The expression of GRP78, p-JNK, caspase-3 at protein levels and GRP78, JNK mRNA were detected by Western blot and RT-PCR.RESULTS:Compared with N group, the number of adherent cells in H group decreased significantly, and cell morphology changed. The expression of OD value in H group decreased obviously (P<0.01), the expression of AI value, GRP78, p-JNK, caspase-3 protein and GRP78, JNK mRNA were significantly increased (P<0.01). HD group compared with H group, the cell damage alleviated, the expression of OD value was increased (P<0.01), the number of apoptosis cells and the AI value in HD group were significantly decreased (P<0.01). Dramatically decreased the expression of p-JNK, caspase-3 protein and JNK mRNA (P<0.01). CONCLUSION: DEX can effectively alleviate A549 cells damage induced by H/R injury, which may be related to inhibition of the JNK pathway.

Key words: JNK, dexmedetomidine, hypoxia/reoxygenation injury, A549 cells, cell apoptosis

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