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中华肝脏外科手术学电子杂志

中华肝脏外科手术学电子杂志 ›› 2017, Vol. 06 ›› Issue (04) : 328 -331. doi: 10.3877/cma.j.issn.2095-3232.2017.04.021

所属专题： 文献；

基础研究

IL-32γ活化Akt通路诱导肝星状细胞LX-2表达MMP2
梁美莲1, 陈小燕2, 张凡1, 潘兴飞3,†()   
  1. 1. 513100　广东省清远市阳山县人民医院感染科
    2. 513100　广东省清远市阳山县人民医院内科
    3. 510150　广州医科大学附属第三医院感染科
  • 收稿日期:2017-04-20 出版日期:2017-08-10
  • 通信作者: 潘兴飞
  • 基金资助:
    国家自然科学基金青年基金(81401306)

IL-32γ induces the expression of MMP2 in hepatic stellate cells LX-2 via activating Akt signaling pathway

Meilian Liang1, Xiaoyan Chen2, Fan Zhang1, Xingfei Pan3,†()   

  1. 1. Department of Infectious Disease, People's Hospital of Yangshan County, Qingyuan 513100, China
    2. Department of Internal Medicine, People's Hospital of Yangshan County, Qingyuan 513100, China
    3. Department of Infectious Disease, the Third Affiliated Hospital of Guangzhou Medical University, Guangzhou 510150, China
  • Received:2017-04-20 Published:2017-08-10
  • Corresponding author: Xingfei Pan
  • About author:
    Corresponding author: Pan Xingfei, Email:
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引用本文：

梁美莲, 陈小燕, 张凡, 潘兴飞. IL-32γ活化Akt通路诱导肝星状细胞LX-2表达MMP2[J]. 中华肝脏外科手术学电子杂志, 2017, 06(04): 328-331.

Meilian Liang, Xiaoyan Chen, Fan Zhang, Xingfei Pan. IL-32γ induces the expression of MMP2 in hepatic stellate cells LX-2 via activating Akt signaling pathway[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2017, 06(04): 328-331.

目的

探讨IL-32γ诱导肝星状细胞LX-2表达基质金属蛋白酶2（MMP2）及其机制。

方法

不同浓度（0、10、20 μg/L）的IL-32γ蛋白与LX-2细胞共培养，RT-PCR检测细胞MMP2 mRNA水平；ELISA检测MMP2蛋白水平，Human Phospho-Akt Immunoassay试剂盒检测活化的Akt水平。不同浓度（0、25、50 μg/L）的重组人活化Akt蛋白与LX-2细胞共培养后，ELISA检测MMP2蛋白；不同浓度（0、10、20 μmol/L）的Akt抑制剂Perifosine与IL-32γ和LX-2细胞共培养后，ELISA检测MMP2蛋白。3组Akt、MMP2 mRNA及蛋白比较采用单因素方差分析和LSD-t检验。

结果

不同浓度的IL-32γ蛋白与LX-2细胞共培养后，MMP2 mRNA和蛋白水平分别为（0.9±0.4）、（5.5±1.4）、（12.2±2.6）和（53±13）、（234±38）、（523±101）μg/L，呈剂量依赖性升高（LSD-t=5.517，3.952和7.855，4.634；P<0.05）。IL-32γ可诱导LX-2细胞Akt表达，依浓度分别为（86±9）、（287±60）、（650±170）RFUs，呈剂量依赖性升高（LSD-t=5.700，3.489；P<0.05）。重组人活化的Akt蛋白可诱导LX-2细胞MMP2表达，依浓度分别为（54±14）、（277±45）、（550±132）μg/L，呈剂量依赖性升高（LSD-t=8.155，3.387；P<0.05）。不同浓度的Perifosine可抑制IL-32γ诱导LX-2细胞的MMP2表达，MMP2蛋白依浓度分别为（553±86）、（233±28）、（89±12）μg/L，呈剂量依赖性降低（LSD-t=6.130，8.083；P<0.05）。

结论

IL-32γ可能通过活化Akt通路诱导肝星状LX-2细胞表达MMP2而参与肝纤维化和肝癌的发生、转移。

关键词: 白细胞介素类, 基质金属蛋白酶2, LX-2细胞
Objective

To investigate the expression of matrix metalloproteinase 2 (MMP2) in hepatic stellate cells LX-2 induced by IL-32γ and its mechanism.

Methods

Different concentrations (0, 10, 20 μg/L) of IL-32γ protein were co-cultured with LX-2 cells, and the expression level of MMP2 mRNA, MMP2 protein and activated Akt was respectively detected by RT-PCR, ELISA and Human Phospho-Akt Immunoassay kit. Different concentrations (0, 25, 50 μg/L) of recombinant human activated Akt protein were respectively co-cultured with LX-2 cells, and different concentrations (0, 10, 20 μmol/L) of Akt inhibitor Perifosine were respectively co-cultured with IL-32γ and LX-2 cells, and the expression of MMP2 protein was detected by ELISA. The expression levels of Akt, MMP2 mRNA and MMP2 protein among three groups were compared using one-way analysis of variance and LSD-t test.

Results

After the co-culture of different concentrations of IL-32γ protein and LX-2 cells, the expression levels of MMP2 mRNA and MMP2 protein were respectively (0.9±0.4), (5.5±1.4), (12.2±2.6) and (53±13), (234±38), (523±101) μg/L, and appeared as dose-dependent elevation (LSD-t=5.517, 3.952 and 7.855, 4.634; P<0.05). The expression of Akt in LX-2 cells could be induced by IL-32γ, the levels was repectively (86±9), (287±60) and (650±170) RFUs in accordance with the concetrations, which appeared as dose-dependent elevation (LSD-t=5.700, 3.489; P<0.05). The expression of MMP2 in LX-2 cells could be induced by recombinant human activated Akt protein, the levels was repectively (54±14), (277±45) and (550±132) μg/L in accordance with the concetrations, which appeared as dose-dependent elevation (LSD-t=8.155, 3.387; P<0.05). Different concentrations of Perifosine could inhibit the inducing effect of IL-32γ upon the expression of MMP2 in LX-2 cells, the expression level of MMP2 protein was respectively (553±86), (233±28) and (89±12) μg/L in accordance with the concetrations, which appeared as dose-dependent reduction (LSD-t=6.130, 8.083; P<0.05).

Conclusions

IL-32γ may induce the expression of MMP2 in hepatic stellate cells LX-2 via activating the Akt signaling pathway, thereby participating in the incidence and metastasis of hepatic fibrosis and liver cancer.

Key words: Interleukins, Matrix metalloproteinase 2, LX-2 cells
表1 不同浓度IL-32γ与LX-2细胞共培养后MMP2表达水平（±s）
IL-32γ蛋白浓度（μg/L） MMP2 mRNA MMP2蛋白（μg/L）
20 12.2±2.6 523±101
10 05.5±1.4 234±380
0 00.9±0.4 053±130
F 33.179 42.729
P <0.05 0<0.05
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