目的探究microRNA-423-5p(miR-423-5p)调节磷脂酰肌醇-3-激酶/丝苏氨酸蛋白激酶(PI3K/AKT)通路在心力衰竭进展中的作用。方法构建大鼠心力衰竭模型,同时用AngⅡ处理(100 n M,24 h)大鼠心肌细胞H9C2构建心力衰竭体外模型。H9C2细胞转染mi...目的探究microRNA-423-5p(miR-423-5p)调节磷脂酰肌醇-3-激酶/丝苏氨酸蛋白激酶(PI3K/AKT)通路在心力衰竭进展中的作用。方法构建大鼠心力衰竭模型,同时用AngⅡ处理(100 n M,24 h)大鼠心肌细胞H9C2构建心力衰竭体外模型。H9C2细胞转染miR-423-5p的抑制剂和模拟剂实现miR-423-5p的敲除和过表达;LY294002用来抑制PI3K/AKT通路的激活。RT-PCR技术检测miR-423-5p的表达; Western blot检测凋亡相关蛋白Bcl-2、Bax、casapse-3/9及PI3K、AKT总蛋白及磷酸化蛋白的表达;流式细胞术检测细胞凋亡。结果 miR-423-5p的表达水平在模型组大鼠心肌组织和AngⅡ处理的H9C2细胞中升高;敲除miR-423-5p可抑制由AngⅡ造成的H9C2细胞凋亡,并促进了p-PI3K及p-AKT的表达,但敲除miR-423-5p的以上作用在PI3K/AKT通路被抑制后全部削弱。结论敲除miR-423-5p可通过激活PI3K/AKT通路抑制心肌细胞凋亡,进而缓解心力衰竭的恶性进展。展开更多
BACKGROUND： It has been found that microRNA-423-5p （miR423-Sp） is an oncogenic factor and frequently upregulated in gastric carcinoma. However, the involvement of miR423- 5p in hepatocellular carcinoma （HCC） has ...BACKGROUND： It has been found that microRNA-423-5p （miR423-Sp） is an oncogenic factor and frequently upregulated in gastric carcinoma. However, the involvement of miR423- 5p in hepatocellular carcinoma （HCC） has been rarely reported. The aim of this study was to assess whether miR423-Sp is aberrantly expressed in HCC tissues, and to characterize its roles in the cancerous biology of HCC.展开更多
主要乳胶蛋白(major latex protein,MLP)是Bet v 1超家族成员,参与植物的各种应激反应。本研究对桑树主要乳胶蛋白基因MaMLP423的启动子pMaMLP423进行了生物信息学分析和克隆表达分析。结果显示pMaMLP423具有启动子必需的TATA-box和CAAT...主要乳胶蛋白(major latex protein,MLP)是Bet v 1超家族成员,参与植物的各种应激反应。本研究对桑树主要乳胶蛋白基因MaMLP423的启动子pMaMLP423进行了生物信息学分析和克隆表达分析。结果显示pMaMLP423具有启动子必需的TATA-box和CAAT-box等基本顺式作用元件,还有参与光响应(Box 4、TCT-motif、GA-motif等)、脱落酸反应(ABRE、AAGAA-motif)、乙烯响应(ERE)、干旱诱导(MYB、MBS等)、创伤响应(WUN-motif)等顺式作用元件。构建植物表达载体pMaMLP423及缺失部分序列的pMaMLP423ΔS1和pMaMLP423ΔS2,转化烟草瞬时表达,qPCR和组织化学染色检测显示,三个启动子均能驱动下游GUS基因的表达,但缺失S1和S2启动子的活性降低。外源激素脱落酸和乙烯利能显著诱导MaMLP4232基因的表达。本研究为深入了解MaMLP423的功能和作用提供了参考。展开更多
文摘目的探究microRNA-423-5p(miR-423-5p)调节磷脂酰肌醇-3-激酶/丝苏氨酸蛋白激酶(PI3K/AKT)通路在心力衰竭进展中的作用。方法构建大鼠心力衰竭模型,同时用AngⅡ处理(100 n M,24 h)大鼠心肌细胞H9C2构建心力衰竭体外模型。H9C2细胞转染miR-423-5p的抑制剂和模拟剂实现miR-423-5p的敲除和过表达;LY294002用来抑制PI3K/AKT通路的激活。RT-PCR技术检测miR-423-5p的表达; Western blot检测凋亡相关蛋白Bcl-2、Bax、casapse-3/9及PI3K、AKT总蛋白及磷酸化蛋白的表达;流式细胞术检测细胞凋亡。结果 miR-423-5p的表达水平在模型组大鼠心肌组织和AngⅡ处理的H9C2细胞中升高;敲除miR-423-5p可抑制由AngⅡ造成的H9C2细胞凋亡,并促进了p-PI3K及p-AKT的表达,但敲除miR-423-5p的以上作用在PI3K/AKT通路被抑制后全部削弱。结论敲除miR-423-5p可通过激活PI3K/AKT通路抑制心肌细胞凋亡,进而缓解心力衰竭的恶性进展。
基金supported by grants from the Fundamental Research Funds for the Central Universities(2015FZA7013)the Natural Science Foundation of Zhejiang Province(LY15H160021)+3 种基金the National Natural Science Foundation of China(81101840 and 81302074)the Medical Science and Technology Project of Zhejiang Province(201472813)the Medical and Health Platform Backbone Personnel Plan of Zhejiang Province(2012RCB015 and 2013KYB107)the Innovative Research Group of the National Natural Science Foundation of China(81421062)
文摘BACKGROUND： It has been found that microRNA-423-5p （miR423-Sp） is an oncogenic factor and frequently upregulated in gastric carcinoma. However, the involvement of miR423- 5p in hepatocellular carcinoma （HCC） has been rarely reported. The aim of this study was to assess whether miR423-Sp is aberrantly expressed in HCC tissues, and to characterize its roles in the cancerous biology of HCC.
