Human placenta-derived mesenchymal stem cell conditioned medium can upregulate BeWo cell viability and zonula occludens expression under hypoxia

doi:10.12307.2021.000

Abstract

Abstract: BACKGROUND: In recent years, mesenchymal stem cells have become a research hotspot in the field of new drug development, and their paracrine effect has also been applied in the repair evaluation experiments of a variety of tissue injured cell models, but there is no research to explore whether their conditioned medium can repair the tight junction function of human placental trophoblast cells under hypoxia.
OBJECTIVE: To demonstrate the effect of mesenchymal stem cell conditioned medium on cell viability and expression of tight junction factors in human placental choriocarcinoma trophoblast cell line BeWo under hypoxia.
METHODS: BeWo cells were exposed to hypoxia induced by 1 000 μmol/L CoCl2 for 12 hours to construct the hypoxia model of placental barrier. For model evaluation, five indexes were measured, including cell viability and expression of hypoxia induced factors-1α, as well as three tight junction factors: zonula occludens 1, claudins 4 and claudins 8. Mesenchymal stem cell conditioned medium was added into hypoxia model to construct hypoxia intervention and reoxygenation groups. To investigate the potential mechanism of zonula occludens 1 expression recovery induced by mesenchymal stem cell conditioned medium, insulin-like growth factor 1 hypoxia intervention and treatment groups were established, and the BeWo cell viability and zonula occludens 1 expression were detected.
RESULTS AND CONCLUSION: (1) BeWo cells viability, zonula occludens 1 and claudins 4 expression were reduced by CoCl2-mimic hypoxia, with hypoxia induced factors-1α mRNA and protein upregulation. However, the expression of claudins 8 mRNA was upregulated, and its protein was downregulated. (2) Human placenta-derived mesenchymal stem cell conditioned medium improved BeWo cells viability and recovered expression level of hypoxia induced factors-1α and zonula occludens 1 under hypoxia induced by CoCl2, but it has no effect on claudins 4. (3) Insulin like growth factor-1 intervention alleviated the inhibition of hypoxia on the viability of BeWo cells, and upregulated the expression of zonula occludens 1.

Key words: stem cells, human placenta-derived mesenchymal stem cells, conditioned medium, CoCl2, hypoxia, BeWo cells, zonula occludens 1

CLC Number:

Lin Qingfan, Xie Yixin, Chen Wanqing, Ye Zhenzhong, Chen Youfang. Human placenta-derived mesenchymal stem cell conditioned medium can upregulate BeWo cell viability and zonula occludens expression under hypoxia[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(在线): 4970-4975.

Figures/Tables 7

References

[1] 邱纪玲,王晓彤,周灏雯,等.间充质干细胞条件培养液修复多种疾病损伤的潜能[J].中国组织工程研究,2019,23(29):4743-4748.
[2] FÖRSTER C. Tight junctions and the modulation of barrier function in disease. Histochem Cell Biol. 2008;130(1):55-70.
[3] LIÉVANO S, ALARCÓN L, CHÁVEZ-MUNGUÍA B, et al. Endothelia of term human placentae display diminished expression of tight junction proteins during preeclampsia. Cell Tissue Res. 2006;324(3):433-448.
[4] 赵海君.缺氧对人胎盘滋养层细胞紧密连接功能的影响及其机制研究[D].南京:南京医科大学,2012.
[5] 唐琳.缺氧对体外滋养细胞及脐静脉内皮细胞中巢蛋白表达的影响[D].吉林:吉林大学,2015.
[6] 张娜娜.地塞米松调控IGF-1表达影响BeWo细胞增殖和合胞体营养转运的机制[D].南京:南京农业大学,2016.
[7] KO JA, MURATA S, NISHIDA T. Up-regulation of the tight-junction protein ZO-1 by substance P and IGF-1 in A431 cells. Cell Biochem Funct. 2009; 27(6):388-394.
[8] 宋颢,章军建.当归对慢性脑低灌注大鼠血脑屏障的保护作用[J].武汉大学学报(医学版),2020,41(5):737-740.
[9] MIYAMOTO N, DE KOZAK Y, JEANNY JC, et al. Placental growth factor-1 and epithelial haemato-retinal barrier breakdown: potential implication in the pathogenesis of diabetic retinopathy. Diabetologia. 2007;50(2):461-470.
[10] CARABALLO JC, YSHII C, BUTTI ML, et al. Hypoxia increases transepithelial electrical conductance and reduces occludin at the plasma membrane in alveolar epithelial cells via PKC-ζ and PP2A pathway. Am J Physiol Lung Cell Mol Physiol. 2011;300(4):L569-578.
[11] 赵海君,钱易,崔毓桂,等.紧密连接的结构与功能[J].国际生殖健康/计划生育杂志,2013,32(5):384-386.
[12] ZHONG X, LUO C, DENG M, et al. Scutellarin-treated exosomes increase claudin 5, occludin and ZO1 expression in rat brain microvascular endothelial cells. Exp Ther Med. 2019;18(1):33-40.
[13] 唐明科.缺氧肺癌细胞释放的腺苷对肺癌脑转移的影响及其可能机制[D].唐山:华北理工大学,2019.
[14] 刘娇阳,王沂峰.紧密连接蛋白Claudin-8的研究进展[J].中国现代医生,2018,56(18):170-174.
[15] MORRIS AP, TAWIL A, BERKOVA Z, et al. Junctional Adhesion Molecules (JAMs) are differentially expressed in fibroblasts and co-localize with ZO-1 to adherens-like junctions. Cell Commun Adhes. 2006;13(4):233-247.
[16] BEUTEL O, MARASPINI R, POMBO-GARCÍA K, et al. Phase Separation of Zonula Occludens Proteins Drives Formation of Tight Junctions. Cell. 2019;179(4):923-936.e11.
[17] ANGELOW S, AHLSTROM R, YU AS. Biology of claudins. Am J Physiol Renal Physiol. 2008;295(4):F867-876.
[18] MAURO L, BARTUCCI M, MORELLI C, et al. IGF-I receptor-induced cell-cell adhesion of MCF-7 breast cancer cells requires the expression of junction protein ZO-1. J Biol Chem. 2001;276(43):39892-39897.