|
|
|
| Human umbilical cord blood mesenchymal stem cell protects apoptosis of AECⅡ induced by LPS via inhibition of MEK/ERK-mediated pathway |
| LIU Jian1 LONG Tingting2 LI Chenyan1 YI Jun1 PAN Ziyang1 YANG Genhui1 HE Xiqiang1 |
1.Department of SICU, the Center Hospital of Xiangtan, Hu′nan Province, Xiangtan 411100, China;
2.Department of Ophthalmology, the Center Hospital of Xiangtan, Hu′nan Province, Xiangtan 411100, China |
|
|
|
Abstract Objective To discuss the effect of human umbilical cord blood mesenchymal stem cells (hUC-MSC) on alveolar type Ⅱ epithelial cells (AEC Ⅱ) apoptosis induce by lipopolysaccharide (LPS), and the role of MEK/ERK signaling pathway. Methods The AECⅡ apoptosis model was established by LPS co-cultured with A549 cells. The apoptosis model was randomly divided into 6 groups according to random number method, they were co-cultured with MSC conditioned medium (MSC-CM), MEK/ERK pathway inhibitor (PD98059) and PBS, which were named A549+LPS+MSC, A549+LPS+PBS, A549+LPS+PD98059, and the negative control groups were A549+PBS, A549+MSC, A549+PD98059. Apoptosis of A549 cells was detected by Annexin V/PI staining flow cytometry 48 h later. MEK/ERK pathway and apoptosis-related signals (BAX/BCL-2, FAS, caspase 3 and caspase 8) were examined by qPCR. The groups of A549+LPS+MSC, A549+LPS+PBS, A549+PBS, and A549+MSC were selected for Western blot detecting. Results The apoptosis model was established by A549+LPS. Compared with A549+LPS+PBS, the apoptosis rate of A549+LPS+MSC and A549+LPS+PD98059 were decreased significantly (P < 0.05). The mRNA expression levels of MEK, ERK, BAX/BCL-2, caspase 3 and caspase 8 in A549+LPS+MSC and A549+LPS+PD98059 were significantly lower than those in A549+LPS+PBS (P < 0.05). The expression of FAS, BAX, cleaved Caspase 3, p-MEK, and p-ERK protein in the A549+LPS+MSC and A549+LPS+PD98059 were significantly lower than A549+LPS+PBS(P < 0.05). There was no significant different in TNF-R1 among each group (P > 0.05). Conclusion hUC-MSCs may inhibit the phosphorylation of MEK/ERK signaling pathway affecting mitochondria pathway and FAS/FASL pathway by paracrine action, thereby attenuating apoptosis of AECⅡ induced by LPS.
|
|
|
|
|
|
[1] Matthay MA，Zemans RL，Zimmerman GA，et al. Acute respiratory distress syndrome [J]. Nat Rev Dis Prim，2019，5（1）：18.
[2] Cecconi M，Evans L，Levy M，et al. Sepsis and septic shock [J]. Lancet，2018，392（10141）：75-87.
[3] 赵锋利，冼绍祥，罗苑苑，等.参附注射液对脓毒症小鼠免疫调节及炎症因子的影响[J].中国医药导报，2019，16（22）：21-25.
[4] Abdulnour R-EE，Gunderson T，Barkas I，et al. Early Intravascular Events Are Associated with Development of Acute Respiratory Distress Syndrome. A Substudy of the LIPS-A Clinical Trial [J]. Am J Respir Crit Care Med，2018，197（12）：1575-1585.
[5] Brauer R，Ge L，Schlesinger SY，et al. Syndecan-1 Attenuates Lung Injury during Influenza Infection by Potentiating c-Met Signaling to Suppress Epithelial Apoptosis [J]. Am J Respir Crit Care Med，2016，194（3）：333-344.
[6] Zhan J，Liu Y，Zhang Z，et al. Effect of penehyclidine hydrochloride on expressions of MAPK in mice with CLP-induced acute lung injury [J]. Mol Biol Rep，2011，38（3）：1909-1914.
[7] Cagnol S，Chambard JC. ERK and cell death：mechanisms of ERK-induced cell death—apoptosis，autophagy and senescence [J]. FEBS J，2010，277（1）：2-21.
[8] 刘坚，吕海金，安玉玲，等.间充质干细胞抑制MEK/ERK信号通路改善盲肠结扎穿孔所致急性肺损伤中肺泡Ⅱ型上皮细胞的凋亡[J].中山大学学报：医学科学版，2016， 37（3）：367-375.
[9] Harrell CR，Sadikot R，Pascual J，et al. Mesenchymal Stem Cell-Based Therapy of Inflammatory Lung Diseases：Current Understanding and Future Perspectives [J]. Stem Cells Int，2019，2019：1-14.
[10] Su VYF，Lin CS，Hung SC，et al. Mesenchymal Stem Cell-Conditioned Medium Induces Neutrophil Apoptosis Associated with Inhibition of the NF-κB Pathway in Endotoxin-Induced Acute Lung Injury [J]. Int J Mol Sci，2019， 20（9）.
[11] Chen CH，Chen YL，Sung PH，et al. Effective protection against acute respiratory distress syndrome/sepsis injury by combined adipose-derived mesenchymal stem cells and preactivated disaggregated platelets [J]. Oncotarget，2017，8（47）：82415-82429.
[12] Salami F，Tavassoli A，Mehrzad J，et al. Immunomodulatory effects of mesenchymal stem cells on leukocytes with emphasis on neutrophils [J]. Immunobiology，2018， 223（12）：786-791.
[13] Ryndak MB，Singh KK，Peng Z，et al. Transcriptional profiling of Mycobacterium tuberculosis replicating in the human type Ⅱ alveolar epithelial cell line，A549 [J]. Genom Data，2015，5：112-114.
[14] 陈刚，廖前德，胡优威，等.成骨诱导人脐带间充质干细胞与纳米羟基磷灰石/聚酰胺66支架材料的生物相容性[J].中国组织工程研究，2012，16（16）：2931-2934.
[15] Huang C，Zheng H，He W，et al. Ghrelin ameliorates the human alveolar epithelial A549 cell apoptosis induced by lipopolysaccharide [J]. Biochem Biophys Res Commun，2016，474（1）：83-90.
[16] Sun Y，Liu WZ，Liu T，et al. Signaling pathway of MAPK/ERK in cell proliferation，differentiation，migration，senescence and apoptosis [J]. J Recept Signal Transduct Res，2015，35（6）：600-604.
[17] Yi J，Gao ZF. MicroRNA-9-5p promotes angiogenesis but inhibits apoptosis and inflammation of high glucose-induced injury in human umbilical vascular endothelial cells by targeting CXCR4 [J]. Int J Biol Macromol，2019， 130：1-9.
[18] Kitai H，Ebi H，Tomida S，et al. Epithelial-to-Mesenchymal Transition Defines Feedback Activation of Receptor Tyrosine Kinase Signaling Induced by MEK Inhibition in KRAS-Mutant Lung Cancer [J]. Cancer Discov，2016，6（7）：754-769.
[19] Schuh K，Pahl A. Inhibition of the MAP kinase ERK protects from lipopolysaccharide-induced lung injury [J]. Biochem Pharmacol，2009，77（12）：1827-1834.
[20] 袁哲，吴思梦，王亚君，等.UC-MSC经PI3K/AKT信号通路影响人肺腺癌A549细胞的凋亡和增殖[J].中国肿瘤生物治疗杂志，2019，26（3）：260-265. |
|
|
|
