Effect of bone marrow mesenchymal stem cells conditioned medium on microglia and its secretion of arginase 1 in rats_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

CHEN Yunhui ¹ , YU Shuguang ² , WU Qiaofeng ² , TANG Yong ² , JIANG Cen ¹ , ZHUANG Zhiqi ² , ZHAO Na ² , HUANG Biao ² ,  XIE Lushuang ^1,2

1. College of Basic Medicine, Chengdu University of Traditional Chinese Medicine, Chengdu Sichuan, 610075, P.R.China;
2. College of Acupuncture and Moxibustion of Chengdu University of Traditional Chinese Medicine, Chengdu Sichuan, 610075, P.R.China;

Corresponding author：

XIE Lushuang, Email: xiescarlett418@163.com

Keywords：

Bone marrow mesenchymal stem cells; microglia; arginase 1; rat

DOI：

10.7507/1002-1892.201710038

Video：

Export PDF Favorites Scan Get Citation

Abstract Full text Figures/Tables Video References Cited by

Objective To observe the effect of bone marrow mesenchymal stem cells (BMSCs) conditioned medium on microglia (MGs) and its secretion of arginase 1 (Arg1). Methods The BMSCs separated through differential adhesion method from the femur and tibia marrow of 4-week-old Sprague Dawley (SD) rats were cultured and identified by Vimentin immunofluorescence staining; whereas MGs separated through trypsin digestion method from the brain of 3-day-old SD rats were cultured and identified by Iba1 immunofluorescence staining. The primary MGs were cultured with DMEM/F12 medium containing BMSCs conditioned medium (experimental group) and with single DMEM/F12 medium (control group), respectively. After 48 hours of culture, the morphology of MGs was observed by inverted phase contrast microscope, the activated state of MGs was detected by using Iba1 immunofluorescence staining, and Arg1 expression of MGs was assessed by Iba1-Arg1 double-labelling immunofluorescence staining and Western blot method. Results Inverted phase contrast microscope observation showed that BMSCs entered logarithmic growth phase at 14 days after culture, and more than 98% cells were positive to Vimentin immunofluorescence staining; whereas MGs entered logarithmic growth phase at 21 days after culture, and around 80% cells were positive to Iba1 immunofluorescence staining. Inverted phase contrast microscope observation displayed that in the experimental group, MGs were activated with increased size of soma, shortened process, and amoeba change. Immunofluorescence staining displayed that the Iba1 positive cells number in the experimental group was significantly higher than that in the control group (t=0.007, P=0.000); double-labelling immunofluorescence staining revealed that the Iba1-Arg1 positive cells number in the experimental group was significantly higher than that in the control group (t=0.007, P=0.000); and Western blot results elucidated that the relative expression of Arg1 protein in the experimental group was significantly higher than that in the control group (t=0.001, P=0.000). Conclusion BMSCs conditioned medium can activate MGs and induce MGs to express Arg1.

Citation： CHEN Yunhui, YU Shuguang, WU Qiaofeng, TANG Yong, JIANG Cen, ZHUANG Zhiqi, ZHAO Na, HUANG Biao, XIE Lushuang. Effect of bone marrow mesenchymal stem cells conditioned medium on microglia and its secretion of arginase 1 in rats. Chinese Journal of Reparative and Reconstructive Surgery, 2017, 31(12): 1500-1505. doi: 10.7507/1002-1892.201710038 Copy

1.	Pittenger MF, Mackay AM, Beck SC, et al. Multilineage potential of adult human mesenchymal stem cells. Science, 1999, 284(5411): 143-147.
2.	Shen Q, Yin Y, Xia QJ, et al. Bone Marrow Stromal Cells Promote Neuronal Restoration in Rats with Traumatic Brain Injury: Involvement of GDNF Regulating BAD and BAX Signaling. Cell Physiol Biochem, 2016, 38(2): 748-762.
3.	Vaquero J, Zurita M. Functional recovery after severe CNS trauma: current perspectives for cell therapy with bone marrow stromal cells. Prog Neurobiol, 2011, 93(3): 341-349.
4.	Ohtaki H,Ylostalo JH, Foraker JE, et al. Stem/progenitor cells from bone marrow decrease neuronal death in global ischemia by modulation of inflammatory/immune responses. Proc Natl Acad Sci U S A, 2008, 105(38): 14638-14643.
5.	Vaquero J, Zurita M. Functional recovery after severe CNS trauma: current perspectives for cell therapy with bone marrow stromal cells. Prog Neurobiol, 2011, 93(3): 341-349.
6.	Liu P, Fleete MS, Jing Y, et al. Altered arginine metabolism in Alzheimer's disease brains. Neurobiol Aging, 2014, 35(9): 1992-2003.
7.	Quirie A, Demougeot C, Bertrand N, et al. Effect of stroke on arginase expression and localization in the rat brain. Eur J Neurosci, 2013, 37(7): 1193-1202.
8.	Bohrnsen F, Godek F, Kiesel J, et al. Influence of TGF-beta1 on tumor transition in oral cancer cell and BMSC co-cultures. J Craniomaxillofac Surg, 2017, 45(5): 731-740.
9.	Shinozaki Y, Shibata K, Yoshida K, et al. Transformation of Astrocytes to a Neuroprotective Phenotype by Microglia via P2Y1 Receptor Downregulation. Cell Rep, 2017, 19(6): 1151-1164.
10.	陈少强, 林建华. 骨髓间充质干细胞移植治疗脊髓损伤的研究现状. 中国修复重建外科杂志, 2007, 21(5): 507-511.
11.	Gao Q, Li Y, Chopp M. Bone marrow stromal cells increase astrocyte survival via upregulation of phosphoinositide 3-kinase/threonine protein kinase and mitogen-activated protein kinase kinase/extracellular signal-regulated kinase pathways and stimulate astrocyte trophic factor gene expression after anaerobic insult. Neuroscience, 2005, 136(1): 123-134.
12.	Shen LH, Li Y, Chopp M. Astrocytic endogenous glial cell derived neurotrophic factor production is enhanced by bone marrow stromal cell transplantation in the ischemic boundary zone after stroke in adult rats. Glia, 2010, 58(9): 1074-1081.
13.	Davalos D, Grutzendler J, Yang G, et al. ATP mediates rapid microglial response to local brain injury in vivo. Nat Neurosci, 2005, 8(6): 752-758.
14.	Colonna M, Butovsky O. Microglia Function in the Central Nervous System During Health and Neurodegeneration. Annu Rev Immunol, 2017, 35: 441-468.
15.	Shinozaki Y, Shibata K, Yoshida K, et al. Transformation of Astrocytes to a Neuroprotective Phenotype by Microglia via P2Y1 Receptor Downregulation. Cell Rep, 2017, 19(6): 1151-1164.
16.	Cizkova D, Devaux S, Le Marrec-Croq F, et al. Modulation properties of factors released by bone marrow stromal cells on activated microglia: an in vitro study. Sci Rep, 2014, 4: 7514.
17.	Ma TC, Barco A, Ratan RR, et al. cAMP-responsive element-binding protein (CREB) and cAMP co-regulate activator protein 1 (AP1)-dependent regeneration-associated gene expression and neurite growth. J Biol Chem, 2014, 289(47): 32914-32925.
18.	Cherry JD, Olschowka JA, O’Banion MK. Arginase 1+ microglia reduce Abeta plaque deposition during IL-1beta-dependent neuroinflammation. J Neuroinflammation, 2015, 12: 203.
19.	Orihuela R, Mcpherson CA, Harry GJ. Microglial M1/M2 polarization and metabolic states. Br J Pharmacol, 2016, 173(4): 649-665.

1. Pittenger MF, Mackay AM, Beck SC, et al. Multilineage potential of adult human mesenchymal stem cells. Science, 1999, 284(5411): 143-147.
2. Shen Q, Yin Y, Xia QJ, et al. Bone Marrow Stromal Cells Promote Neuronal Restoration in Rats with Traumatic Brain Injury: Involvement of GDNF Regulating BAD and BAX Signaling. Cell Physiol Biochem, 2016, 38(2): 748-762.
3. Vaquero J, Zurita M. Functional recovery after severe CNS trauma: current perspectives for cell therapy with bone marrow stromal cells. Prog Neurobiol, 2011, 93(3): 341-349.
4. Ohtaki H,Ylostalo JH, Foraker JE, et al. Stem/progenitor cells from bone marrow decrease neuronal death in global ischemia by modulation of inflammatory/immune responses. Proc Natl Acad Sci U S A, 2008, 105(38): 14638-14643.
5. Vaquero J, Zurita M. Functional recovery after severe CNS trauma: current perspectives for cell therapy with bone marrow stromal cells. Prog Neurobiol, 2011, 93(3): 341-349.
6. Liu P, Fleete MS, Jing Y, et al. Altered arginine metabolism in Alzheimer's disease brains. Neurobiol Aging, 2014, 35(9): 1992-2003.
7. Quirie A, Demougeot C, Bertrand N, et al. Effect of stroke on arginase expression and localization in the rat brain. Eur J Neurosci, 2013, 37(7): 1193-1202.
8. Bohrnsen F, Godek F, Kiesel J, et al. Influence of TGF-beta1 on tumor transition in oral cancer cell and BMSC co-cultures. J Craniomaxillofac Surg, 2017, 45(5): 731-740.
9. Shinozaki Y, Shibata K, Yoshida K, et al. Transformation of Astrocytes to a Neuroprotective Phenotype by Microglia via P2Y1 Receptor Downregulation. Cell Rep, 2017, 19(6): 1151-1164.
10. 陈少强, 林建华. 骨髓间充质干细胞移植治疗脊髓损伤的研究现状. 中国修复重建外科杂志, 2007, 21(5): 507-511.
11. Gao Q, Li Y, Chopp M. Bone marrow stromal cells increase astrocyte survival via upregulation of phosphoinositide 3-kinase/threonine protein kinase and mitogen-activated protein kinase kinase/extracellular signal-regulated kinase pathways and stimulate astrocyte trophic factor gene expression after anaerobic insult. Neuroscience, 2005, 136(1): 123-134.
12. Shen LH, Li Y, Chopp M. Astrocytic endogenous glial cell derived neurotrophic factor production is enhanced by bone marrow stromal cell transplantation in the ischemic boundary zone after stroke in adult rats. Glia, 2010, 58(9): 1074-1081.
13. Davalos D, Grutzendler J, Yang G, et al. ATP mediates rapid microglial response to local brain injury in vivo. Nat Neurosci, 2005, 8(6): 752-758.
14. Colonna M, Butovsky O. Microglia Function in the Central Nervous System During Health and Neurodegeneration. Annu Rev Immunol, 2017, 35: 441-468.
15. Shinozaki Y, Shibata K, Yoshida K, et al. Transformation of Astrocytes to a Neuroprotective Phenotype by Microglia via P2Y1 Receptor Downregulation. Cell Rep, 2017, 19(6): 1151-1164.
16. Cizkova D, Devaux S, Le Marrec-Croq F, et al. Modulation properties of factors released by bone marrow stromal cells on activated microglia: an in vitro study. Sci Rep, 2014, 4: 7514.
17. Ma TC, Barco A, Ratan RR, et al. cAMP-responsive element-binding protein (CREB) and cAMP co-regulate activator protein 1 (AP1)-dependent regeneration-associated gene expression and neurite growth. J Biol Chem, 2014, 289(47): 32914-32925.
18. Cherry JD, Olschowka JA, O’Banion MK. Arginase 1+ microglia reduce Abeta plaque deposition during IL-1beta-dependent neuroinflammation. J Neuroinflammation, 2015, 12: 203.
19. Orihuela R, Mcpherson CA, Harry GJ. Microglial M1/M2 polarization and metabolic states. Br J Pharmacol, 2016, 173(4): 649-665.

Chinese Journal of Reparative and Reconstructive Surgery

Effect of bone marrow mesenchymal stem cells conditioned medium on microglia and its secretion of arginase 1 in rats

Abstract Full text Figures/Tables Video References Cited by

Previous Article

Next Article

Format

Content