Effect of Microencapsulation on Expression of Osmoregulation Genes and Exogenous Regulation in HepG2 Cell
1 Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China
2 Department of Public Health, NanTong University, Nantong 226019, China
Abstract:The aim of this work is to investigate the influence of microencapsulation on the expression of the osmoregulation genes and exogenous regulation in HepG2 cells. We compared the expression of sodium/myoinositol cotransporter (SMIT) and taurine transporter (TAUT) in HepG2 cells under different cultured conditions through realtime RTPCR. The effects of exogenous antiosmotic stress matter on cell viability and albumin level were also evaluated through MTT assay and ELISA assay. We did not find a significant difference in the expression of SMIT. However, the results showed that the expression level of TAUT in encapsulated cells was approximately 4.3 times higher than that of monolayer cells(P<0.01). Accordingly, we found that taurine at 1~1.5 mM significantly increased the viability by 15%~30% and the biosynthetic function by 15% in microencapsulated HepG2 cells(P<0.05). Trehalose, as for it, did not alter the biosynthetic function but increased the viability of the encapsulated cells by 20%~30% at 0.1 mM(P<0.05). In conclusion, osmotic stress exists in the microcapsules and affects genes expression. Exogenous antiosmotic stress matter can prevent the inhibition effects of hyperosmotic stress on cellular growth.
[1]Li Yuanqiu, Wang Jian, Wen Xinyu, et al. Transplantation of comicroencapsulated hepatocytes and HUVECs for treatment of fulminant hepatic failure [J]. Int J Artif Organs, 2012, 35(6):458-465.
[2]Li Shuangyue, Sun Zhijie, Lv Guojun, et al. Microencapsulated umbilical cord blood cells repair mouse hepatic injury by intraperitoneal transplantation [J]. Cytotherapy, 2010, 11(8):1032-1040.
[3]Addo RT, Siddig A, Siwale R, et al. Formulation, characterization and testing of tetracaine hydrochlorideloaded albuminchitosan microparticles for ocular drug delivery [J]. J Microencapsul, 2010, 27(2):95-104.
[4]Sun Zhijie, Lv Guojun, Li Shuangyue, et al. Probing the role of microenvironment for microencapsulated Sacchromyces cerevisiae under osmotic stress [J]. J Biotechnol, 2007, 128(1):150-161.
[5]Ning Xiaojuan, Sun Zhijie, Zhong Shen, et al. Growth and metabolism of osmosensentive yeast Y02724 and highosmotic resistant yeast Hansel in alginatechitosanalginate microcapsules [J]. Sheng Wu Gong Cheng Xue Bao, 2008, 24(7):1274-1278.
[6]肖静, 张英, 于炜婷, 等. 微囊化HepG2细胞脂代谢变化对线粒体功能和蛋白合成能力的影响 [J]. 中国组织工程研究与临床康复,2010,14(38):7082-7086.
[7]Walker J, Martin C, Callaghan R. et al. Inhibition of Pglycoproteinfunction by XR9576 in a solid tumour model can restore anti cancer drug efficacy [J]. Eur J Cancer, 2004, 40(4):594- 605.
[8]Hilge RB, Rehm HJ. Comparison of fermentation properties and speci fic enzyme activities of free and calciumalginateentrapped Saccharomyces cerevisiae [J]. Appl Microbiol Biotechnol, 1990, 33(7):54-58.
[9]Norton S, Amore DT. Physiological effects of yeast cell immobilization: applications for brewing [J]. Enzyme Microb Technol, 1994, 16(1):365-375.
[10]Palkova Z, Janderova B, Gabriel J, et al. Ammonia mediates communication between yeast colonies [J]. Nature, 1997, 390(2): 532-536.
[11]Maurice BB, Joan DF, Natallaid ID, et al. Cellular Response to Hyperosmotic Stresses [J]. Physiol Rev, 2007, 87(1):1441-1474.
[12]Steffann HO. Intracellular water homeostasis and the mammalian cellular osmotic stress response [J]. J Cell Phy, 2006, 206(1):9-15.
[13]Akinlzhanov A,Krenacs L,Schlegel T, et al.Epigenetic changes and suppression of the nuclear factor of activated T cell (NFATCl) promoter in human lymphomas with defects in immunoreceptor signaling [J].Am J Pathol, 2008, 172(1):215-224.
[14]Takashi I, Yasushi F, Mayo H, et al. Expression of taurine transporter is regulated through the TonE (tonicity responsive element)/TonEBP (TonE binding protein) pathway and contributes to cytoprotection in HepG2 cells [J]. Biochem J, 2004, 382(Pt 1):177-182.
[15]Warskulat U, Reinen A, Grether BS, et al. The osmolyte strategy of normal human keratinocytes in maintaining cell homeostasis [J]. J Invest Dermatol, 2004, 123(3):516-521.
[16]Hideo S, Eriko T, Yu HK, et al. Functional characterization and regulation of the taurine transporter and cysteine dioxygenase in human hepatoblastoma HepG2 cells [J]. Biochem J, 2003, 375(Pt 2):441-447.
[17]Hijab S, Havalad S, Snyder AK. The role of organic osmolytes in the response of cultured astrocytes to hyperosmolarity [J]. Am J Ther, 2011, 18(5):366-370.
[18]Wikstrm J, Elomaa M, Nevala L, et al. Viability of freeze dried microencapsulated human retinal pigment epithelial cells [J]. Eur J Pharm Sci, 2012, 47(2):520-526.
[19]Sukit L, Yifei W, Amadeu K, et al. Experimental and computational studies investigating trehalose protection of HepG2 cells from palmitateinduced toxicity [J]. Biophys J, 2008, 94(7):2869-2883.