Tissue Inhibitor of Matrix Metalloproteinase 2-Loaded Polylactic Acid-Polyethylene Glycol Nanoparticles Directly Regulates Renal Tubular Epithelial Cell Proliferation via Protein Tyrosine Kinase KIT
Tissue Inhibitor of Matrix Metalloproteinase 2-Loaded Polylactic Acid-Polyethylene Glycol Nanoparticles Directly Regulates Renal Tubular Epithelial Cell Proliferation via Protein Tyrosine Kinase KIT
复制标题
负载基质金属蛋白酶2的组织抑制剂聚乳酸-聚乙二醇纳米颗粒通过蛋白酪氨酸激酶试剂盒直接调节肾小管上皮细胞增殖
DOI：
10.1166/jnn.2020.18717
复制
发表时间：
2020
影响因子：
--
通讯作者：
Ling Yunzhi
中科院分区：
文献类型：
--
作者：
Ke Xiaosu;Zhou Qin;Li Bin;Shao Xiaofei;Hong Guobao;Zou Hequn;Ling Yunzhi
TIMP 2与急性肾损伤（阿基）相关，但其机制尚不清楚。因此，本研究探讨TIMP 2在人肾小管上皮细胞HK-2细胞中的调控。TIMP 2在正常表达水平下作用于HK-2细胞后，检测细胞增殖情况。应用GST pulldown和质谱技术研究TIMP 2的相互作用蛋白，并通过免疫荧光实验确定TIMP 2在HK-2细胞中的定位。与对照组和BB-94处理组相比，TIMP 2过表达组的细胞存活率、CCND 1、C-FOS、MAPK 1和P-MAPK 1的表达水平均显著降低（P < 0.05）。KIT是TIMP 2的相互作用蛋白，定位于HK-2细胞的胞浆和胞膜。与对照组相比，抑制KIT和过表达TIMP 2均能显著抑制细胞增殖，并降低CCND 1、MAPK 1和P-MAPK 1的表达水平（P < 0.05）。TIMP 2过表达组与KIT抑制组细胞增殖及上述蛋白表达水平无明显差异。结果显示，TIMP 2通过KIT降低CCND 1、MAPK 1和P-MAPK 1的表达水平来调节细胞增殖，表明TIMP 2在阿基中直接调节细胞增殖而不抑制基质金属蛋白酶。此外，PLA-PEG纳米颗粒成功地将TIMP 2转运到靶点，对细胞增殖没有显著影响。
TIMP2 has been previously reported to be associated with acute kidney injury (AKI); however, the underlying mechanism remains unclear. Therefore, the present study investigated the regulation of TIMP2 in human tubular epithelial cells HK-2 cells. Proliferation of HK-2 cells treated by TIMP2 at normal expression level was detected. GST pulldown and mass spectrometry were performed to investigate the interacting protein of TIMP2 and immunofluorescence test was used to determine the location of the protein in HK-2 cells. Cell viability as well as the expression level of CCND1, C-FOS, MAPK1 and P-MAPK1 were detected in the samples treated by overexpressed TIMP2 and the inhibitor of the interacting protein KM TIMP2 significantly inhibited cell proliferation compared with the control and BB-94-treated groups (P < 0.05). KIT was identified as the interacting protein of TIMP2, and was located in both the cytoplasm and membrane of HK-2 cells. Inhibited KIT and the overexpressed of TIMP2 both significantly suppressed cell proliferation and decreased the expression levels of CCND1, MAPK1, and P-MAPK1 compared with the control (P < 0.05). No significant difference was observed in cell proliferation and the expression level of aforementioned proteins between overexpressed TIMP2 and KIT-inhibited group. The results revealed that TIMP2 regulates cell proliferation by reducing the expression levels of CCND1, MAPK1, and P-MAPK1 via KIT, indicating that TIMP2 directly regulates cell proliferation without inhibiting matrix metalloproteinases in AKI. Furthermore, PLA-PEG nanoparticles successfully transported TIMP2 to the target with no significant effect on cell proliferation.