The protective effects of amniotic mesenchymal stem cells for rats early myocardial injury induced by severe burns
MOU Huan1 LIU Yang2▲ LIU Jian3 ZHOU Xinli1 QIANG Tinghui1 DU Xingguo1 DING Chen1 XUE Chao1
1.The First Department of Orthopaedic Surgery, Hanzhong Central Hospital, Shaanxi Province, Hanzhong 723000, China;
2.Burn Center of People's Liberation Army, the First Affiliated Hospital of the Fourth Military Medical University, Shaanxi Province, Xi'an 710032, China;
3.Department of Neurosurgery, the First Affiliated Hospital of the Fourth Military Medical University, Shaanxi Province, Xi'an 710032, China
Abstract:Objective To explore the effects and related mechanism of amniotic mesenchymal stem cells (AMSCs) for rats myocardial injury induced by severe burns. Methods The AMSCs were obtained by the method of collagenase-pancreatic enzyme digestion. Biological markers were assayed with flow cytometry. The activity of AMSCs was identified using adipogenic and osteoplastic differentiation. 24 healthy male SD rats were randomly divided into sham group, burn group and AMSCs group. The dorsal skin of rats in burn group and AMSCs group were exposed to 98℃ water for 15 s which led to the third degree, 30% TBSA burns, while the rats in the sham group were exposed to 37℃ water for 15 s. Rats in burn group and AMSCs group were given 0.9% 10 mL (50 mL/kg) saline intraperitoneally immediately after burns. PBS or AMSCs were injected through caudal vein 3 hours later. 48 hours later, rats were sacrificed and the myocardium and blood from aorta abdominalis were collected. The levels of creatine kinase (CK), lactic dehydrogenase (LDH) were detected by ELISA. The mRNA levels of caspase-3, TNF-α, IL-1β and IL-10 were detected by RT-PCR. The data were analyzed through AVONA and LSD-t test by SPSS. Results The positive rates of CD29, CD44, CD90, CDl05 were very high in the third generation of AMSCs, however, the positive rate of CD34 was low. AMSCs underwent osteogenic and adipogenic differentiation after induction. The levels of CK and LDH in rats of the burn group were significantly increased compared with the sham group (P < 0.05). The levels of CK and LDH in AMSCs group were significantly lower than those in the burn group (P < 0.05). The mRNA levels of caspase-3, IL-1β and TNF-α in the myocardium of the burn group were significantly increased compared with the sham group (P < 0.05). The mRNA levels of caspase-3, IL-1β and TNF-α in myocardium of the AMSCs group were significantly lower than those in the burn group (P < 0.05). The mRNA level of IL-10 in the burn group was significantly lower than that in the sham group (P < 0.05). In the AMSCs group, the expression of IL-10 mRNA was significantly higher than that in the burn group (P < 0.05). Conclusion AMSCs can protect against myocardial injury induced by severe burns. The levels of CK and LDH decreased, indicating that the injury of myocardial decreased. During which, the inflammatory factors, IL-1β and TNF-α mRNA decreased and anti-inflammatory factor IL-10 mRNA increased.
[1] Weil BR,Markel TA,Herrmann JL,et al. Mesenchymal stem cells enhance the viability and proliferation of human fetal intestinal epithelial cells following hypoxic injury via paracrine mechanisms [J]. Surgery,2009,146(2):190-197.
[2] 谢松涛,樊磊,杨龙龙,等.脂肪间充质干细胞对严重烧伤大鼠早期心肌损伤的保护作用[J].中华损伤与修复杂志电子版,2015,10(4):4-8.
[3] Méndez-Ferrer S,Michurina TV,Ferraro F,et al. Mesenchymal and haematopoietic stem cells form a uniquebone marrow niche [J]. Nature,2010,466(7308):829-834.
[4] Insausti CL,Blanquer M,Bleda P,et al. The amniotic membrane as a source of stem cells [J]. Histol Histopathol,2010, 25(1):91-98.
[5] Koh JW,Shin YJ,Oh JY,et al. The expression of TIMPs in cryo-preserved and freeze-dried amniotic membrane [J]. Curr Eye Res,2007,32(7-8):611-616.
[6] Vosdoganes P,Hodges RJ,Lim R,et al. Human amnion epithelial cells as a treatment for inflammation-induced fetal lung injury in sheep [J]. Am J Obstet Gynecol,2011,205(2):15626-1533.
[7] Hu X,Xu Y,Zhong Z,et al. A Large-Scale Investigation of Hypoxia-Preconditioned Allogeneic Mesenchymal Stem Cells for Myocardial Repair in Nonhuman Primates:Paracrine Activity Without Remuscularization [J]. Circ Res,2016,118(6):970-983.
[8] Takahashi K,Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors [J]. Cell,2006,126(4):663-676.
[9] 黄跃生.烧伤后早期心肌损害的分子机制及防治研究进展[J].中华烧伤杂志,2004,20(5):257-259.
[10] Nguyen PK,Rhee JW,Wu JC. Adult stem cell therapy and heart failure,2000 to 2016:a systematic review [J]. JAMA Cardiol,2016,1(7):831.
[11] Ge X,Wang IN,Toma I,et al. Human amniotic mesenchymal stem cell-derived induced pluripotent stem cells may generate a universal source of cardiac cells [J]. Stem Cells Dev,2012,21(15):2798-2808.
[12] Song YS,Joo HW,Park IH,et al. Transplanted Human Amniotic Epithelial Cells Secrete Paracrine Proangiogenic Cytokines in Rat Model of Myocardial Infarction [J]. Cell Transplant,2015,24(10):2055-2064.
[13] 张家平,黄跃生,周新,等.严重烫伤大鼠心肌细胞凋亡与心功能损害的关系[J].中华烧伤杂志,2002,18(5):272-275.
[14] Toda A,Okabe M,Yoshida T,et al. The potential of amniotic membrane/amnion-derived cells for regeneration of various tissues [J]. J Pharmacol Sci,2007,105(3):215-228.
[15] Perrini C,Strillacci MG,Bagnato A,et al. Microvesicles secreted from equine amniotic-derived cells and their potential role in reducing inflammation in endometrial cells in an in-vitro model [J]. Stem Cell Res Ther,2016, 7(1):169.
[16] Angelini A,Castellani C,Ravara B,et al. Stem-cell therapy in an experimental model of pulmonary hypertension and right heart failure: role of paracrine and neurohormonal milieu in the remodeling process [J]. J Heart Lung Transplant,2011, 30(11):1281-1293.
[17] Murphy S,Lim R,Dickinson H,et al. Human amnion epithelial cells prevent bleomycin-induced lung injury and preserve lung function [J]. Cell Transplant,2011,20(6):909-923.