[1]李茂华,滕苗.严重烧伤大鼠血清对骨髓间充质干细胞迁移的影响[J].第三军医大学学报,2015,37(16):1643-1647.
 Li Maohua,Teng Miao.Serum from severely burned rats promotes migration in rat bone marrow mesenchymal stem cells through PI3K/Akt signal pathway[J].J Third Mil Med Univ,2015,37(16):1643-1647.
点击复制

严重烧伤大鼠血清对骨髓间充质干细胞迁移的影响(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
37卷
期数:
2015年第16期
页码:
1643-1647
栏目:
论著
出版日期:
2015-08-30

文章信息/Info

Title:
Serum from severely burned rats promotes migration in rat bone marrow mesenchymal stem cells through PI3K/Akt signal pathway
作者:
李茂华滕苗
重庆医科大学附属第一医院烧伤整形外科
Author(s):
Li Maohua Teng Miao

Department of Burn and Plastic Surgery, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China

关键词:
烧伤骨髓间充质干细胞迁移PI3K/Akt信号通路
Keywords:
burns bone marrow mesenchymal stem cells migration PI3K/Akt signal pathway
分类号:
R329.28;R331.22;R644
文献标志码:
A
摘要:

目的      观察严重烧伤大鼠血清刺激下骨髓间充质干细胞(BMSCs)迁移能力变化,探讨PI3K/Akt信号通路在BMSCs细胞迁移中的作用。      方法      建立严重烧伤大鼠模型,制备烧伤大鼠血清。①用体积分数10%的烧伤大鼠血清刺激BMSCs细胞不同时间后,Transwell小室检测BMSCs细胞迁移能力,Western blot检测BMSCs细胞内磷酸化p38(p-p38)和磷酸化Akt(p-Akt)表达水平。②另取BMSCs细胞随机分为4组:对照组(含10%胎牛血清)、烧伤血清组(含10%烧伤大鼠血清)、烧伤血清+抑制剂组(含10%烧伤大鼠血清+10 μmol/L的LY294002)、烧伤血清+激动剂组(含10%烧伤大鼠血清+200 ng/mL的IGF-1),Transwell小室检测各组细胞迁移情况。      结果      ①烧伤血清作用0、1、3、6、12、24 h时,BMSCs细胞较迁移数分别为0、1.0±0.7、14.6±4.6、45.0±6.4、82.8±8.3、122.2±11.4,3 h时BMSCs细胞出现迁移,6、12、24 h时BMSCs细胞数量多于3 h(P<0.01);烧伤血清作用0、1、3、6、12、24 h时,BMSCs细胞p-p38水平分别为1.02±0.50、1.11±0.17、1.04±0.23、1.02±0.20、1.16±0.22、1.00±0.19,p-p38蛋白表达1、3、6、12、24 h变化趋势不明显,差异无统计学意义(P>0.05);而p-Akt水平分别为0.59±0.08、0.65±0.06、0.83±0.22、1.10±0.21、1.22±0.19、1.62±0.30,6、12、24 h 时较作用0 h时(加入血清后即刻)水平明显增高(P<0.05);12、24 h时较3 h时明显增高(P<0.05)。②对照组、烧伤血清组、烧伤血清+抑制剂组、烧伤血清+激动剂组培养24 h后,烧伤血清组BMSCs细胞迁移数量较对照组增多(P<0.01),与烧伤血清组比较,烧伤血清+激动剂组BMSCs细胞迁移数量增多(P<0.01),烧伤血清+抑制剂组细胞迁移数量明显减少(P<0.01)。      结论      严重烧伤大鼠血清可通过PI3K/Akt信号通路促进BMSCs细胞迁移。

Abstract:

Objective      To observe the migration ability of bone marrow mesenchymal stem cells (BMSCs) after the treatment of the serum derived from severely burned rats, and determine the role of PI3K/Akt signaling pathway in the process.       Methods      After severely burned rat model was established, the rat serum was derived. (1) After the BMSCs were treated by rat serum at volume fraction of 10% for different time periods, Transwell chambers test was used to detect the migration of BMSCs, and Western blotting to measure the expression of intracellular phosphorylation p38 (p-p38) and phosphorylated Akt (p-Akt). (2) BMSCs cells were randomly divided into 4 groups of cells, that is, control group (treated by 10% fetal bovine serum), serum group (containing 10% serum from burned rats in general rat serum), serum +inhibitor group (containing 10% serum from burned rats +10 μmol/L LY294002), serum +agonist group (containing 10% serum from burned rats +200 ng/mL IGF-1). Transwell chamber test was used to detect cell migration.       Results      (1) In 0, 1, 3, 6, 12 and 24 h after burned serum treatment, the quantity of BMSCs cells migration was 0, 1.0±0.7, 14.6±4.6, 45.0±6.4, 82.8±8.3 and 122.2±11.4 respectively. Cell migration was observed since 3 h, the number of migrated cells in 6, 12 and 24 h was significantly larger than that of 3 h (P<0.01). After burned serum treatment for 0, 1, 3, 6, 12 and 24 h, the expression level of p-p38 was 1.02±0.50, 1.11±0.17, 1.04±0.23, 1.02±0.2, 1.16±0.22 and 1.00±0.19 respectively, and no significant difference was seen in the expression levels among these time points (P>0.05). The expression level of p-Akt was 0.59±0.08, 0.65±0.06, 0.83±0.22, 1.10±0.21, 1.22±0.19 and 1.62±0.30 respectively for 6, 12 and 24 h treatment, and the levels were significantly higher than that of 0 h (instantly after serum adding, P<0.05), and that of 12 and 24 h was significantly higher than that of 3 h (P<0.05). (2) In the control group, burned serum group, the burned serum+ inhibitors, burned serum+ agonist group after 24 hours’ treatment, the number of BMSCs migration was larger in the burned serum group than the control group (P<0.01), in the burned serum +agonist group than the burned serum group (P<0.01), and was the lowest in the burned serum+ inhibitor group (P<0.01).       Conclusion      The serum derived from severe burned rats promotes the migration of BMSCs by PI3K/Akt signal pathway.
 

参考文献/References:

[1]黄跃生. 烧伤后早期心肌损害与防治[J]. 中华烧伤杂志, 2008, 24(5): 369-371.
[2]Hare J M, Fishman J E, Gerstenblith G, et al.  Comparison of allogeneic vs autologous bone marrow-derived mesenchymal stem cells delivered by transendocardial injection in patients with ischemic cardiomyopathy: the POSEIDON randomized trial[J].  JAMA, 2012, 308(22): 2369-2379.
[3]李娜, 胡大海, 王耀军, 等.  脂肪间充质干细胞对烧伤脓毒症小鼠肾脏损伤的作用[J]. 中华烧伤杂志, 2013, 29(3): 249-254.
[4]Abazov V M, Abbott B, Acharya B S, et al.  Precision measurement of the ratio B(t→Wb)/B(t→Wq) and extraction of V(tb)[J].  Phys Rev Lett, 2011, 107(12): 121802.
[5]Chim H, Miller E, Gliniak C, et al.  Stromal-cell-derived factor (SDF) 1-alpha in combination with BMP-2 and TGF-beta1 induces site-directed cell homing and osteogenic and chondrogenic differentiation for tissue engineering without the requirement for cell seeding[J].  Cell Tissue Res, 2012, 350(1): 89-94.
[6]郑军, 黄跃生, 黄晓元, 等.  反义p38α基因转染对缺氧复合烧伤血清处理心肌细胞炎性因子的表达[J].  第三军医大学学报, 2004, 26(23): 2097-2101.
[7]Leclerc T, Thepenier C, Jault P, et al.  Cell therapy of burns[J].  Cell Prolif, 2011, 44(Suppl 1): 48-54.
[8]Maxson S, Lopez E A, Yoo D, et al.  Concise review: role of mesenchymal stem cells in wound repair[J].  Stem Cells Transl Med, 2012, 1(2): 142-149.
[9]Strauer B E, Steinhoff G.  10 years of intracoronary and intramyocardial bone marrow stem cell therapy of the heart: from the methodological origin to clinical practice[J].  J Am Coll Cardiol, 2011, 58(11): 1095-1104.
[10]Chen L, Qiu R, Xu Q.  Stem cell therapy for ischemic stroke[J].  J Nanosci Nanotechnol, 2014, 14(1): 976-982.
[11]Gupta N K, Armstrong E J, Parikh S A.  The current state of stem cell therapy for peripheral artery disease[J].  Curr Cardiol Rep, 2014, 16(2): 447.
[12]Ito H.  Chemokines in mesenchymal stem cell therapy for bone repair: a novel concept of recruiting mesenchymal stem cells and the possible cell sources[J].  Mod Rheumatol, 2011, 21(2): 113-121.
[13]Hare J M, Difede D, Heldman A W.  Use of stem cells for ischemic cardiomyopathy--reply[J].  JAMA, 2013, 309(14): 1458-1459.
[14]Cho Y H, Cha M J, Song B W, et al.  Enhancement of MSC adhesion and therapeutic efficiency in ischemic heart using lentivirus delivery with periostin[J].  Biomaterials, 2012, 33(5): 1376-1385.
[15]Scrima M, De-Marco C, Fabiani F, et al.  Signaling networks associated with AKT activation in non-small cell lung cancer (NSCLC): new insights on the role of phosphatydil-inositol 3 kinasel[J].PLoS One, 2012, 7(2): e30427.
[16]Ballard-Croft C, White D J, Maass D L, et al.  Role of p38 mitogen-activated protein kinase in cardiac myocyte secretion of the inflammatory cytokine TNF-alpha[J].  Am J Physiol Heart Circ Physiol, 200l, 280(5): H1970-H1981.
[17]宋华培, 黄跃生, 党永明, 等.  PI3K/Akt信号途径抑制烧伤后大鼠缺血缺氧心肌细胞凋亡[J].  第三军医大学学报, 2009, 31(1): 52-55.
[18]韩冰, 付小兵, 梁雪梅, 等.  烧伤大鼠血清诱导骨髓间充质干细胞分化为表皮细胞和血管内皮细胞的实验研究[J].  解放军医学杂志, 2004, 29(8): 652-654.
[19]Li Y, Yu X, Lin S, et al.  Insulin-like growth factor 1 enhances the migratory capacity of mesenchyamal stem cells[J].  Biochem Biophys Res Commun, 2007, 356(3): 780-784.
 

相似文献/References:

[1]张蕾,陈沅,田杰,等.心肌细胞介导骨髓间充质干细胞的心肌样分化[J].第三军医大学学报,2005,27(16):1681.
[2]王颖楠,范雪梅,赵敏,等.脱细胞膀胱基质复合大鼠骨髓间充质干细胞体外构建组织工程化吊带治疗压力性尿失禁的初步研究[J].第三军医大学学报,2012,34(22):2269.
 Wang Yingnan,Fan Xuemei,Zhao Min,et al.Preliminary study on the construction of a tissue-engineered sling with BMSCs and UBM in vitro for treating stress urinary incontinence[J].J Third Mil Med Univ,2012,34(16):2269.
[3]周长立,任先军,蒋涛,等.Wnt7a基因对大鼠骨髓间充质干细胞增殖及向神经元样细胞分化的影响[J].第三军医大学学报,2013,35(08):702.
 Zhou Changli,Ren Xianjun,Jiang Tao,et al.Wnt7a gene stimulates mesenchymal stem cell proliferation and differentiation into neuron-like cells[J].J Third Mil Med Univ,2013,35(16):702.
[4]郝磊,邹仲敏,王军平,等.hPDGF-A/hBD2双基因共表达腺病毒载体的构建及表达[J].第三军医大学学报,2007,29(10):859.
 HAO Lei,ZOU Zhong-min,WANG Jun-ping,et al.Construction and identification of recombinant adenovirus expressing hPDGF-A and hBD2[J].J Third Mil Med Univ,2007,29(16):859.
[5]郭书权,吴雪晖,许建中,等.两种方法分离小型猪骨髓间充质干细胞的比较[J].第三军医大学学报,2007,29(10):988.
[6]黄文秋,黄宏,徐祥,等.mTOR及其下游信号通路在骨髓间充质干细胞氧化应激损伤中的变化及作用[J].第三军医大学学报,2013,35(02):114.
 Huang Wenqiu,Huang Hong,Xu Xiang,et al.Changes and roles of mTOR and its downstream signaling passway in mouse bone marrow stem cells with oxidative stress injury[J].J Third Mil Med Univ,2013,35(16):114.
[7]冯一梅,徐辉,邹仲敏,等.hPDGF-A/hBD2双基因转染对大鼠骨髓间充质干细胞生物学特性的影响[J].第三军医大学学报,2008,30(06):472.
 FENG Yi-mei,XU Hui,ZOU Zhong-min,et al.Effects of hPDGF-A/hBD2 genes transfection on rat bone marrow mesenchymal stem cells[J].J Third Mil Med Univ,2008,30(16):472.
[8]姚青,宋治远,马显光.脉冲微交流电刺激促进体外诱导大鼠骨髓间充质干细胞向心肌分化[J].第三军医大学学报,2008,30(05):410.
 YAO Qing,SONG Zhi-Yuan,MA Xian-guang.Electrical stimulation promotes the differentiation of rat bone marrow mesenchymal stem cells to cardiomyocyte induced by 5-azacytidine in vitro[J].J Third Mil Med Univ,2008,30(16):410.
[9]朱淑霞,李永华,宋治远,等.电磁场促进骨髓间充质干细胞体外诱导分化时细胞增殖[J].第三军医大学学报,2008,30(05):421.
 ZHU Shu-xia,LI Yong-hua,SONG Zhi-yuan,et al.Pulsed electromagnetic fields improve proliferation of rat marrow mesenchymal stem cells induced by 5-azacytidine[J].J Third Mil Med Univ,2008,30(16):421.
[10]金旭红,杨柳,段小军,等.体外纳米磁标记骨髓间充质干细胞生物学特性及其MR成像[J].第三军医大学学报,2008,30(04):275.
 JIN Xu-hong,YANG Liu,DUAN Xiao-jun,et al.Biological characteristics and in vitro MRI of superparamagnetic ironoxide labeled bonederived mesenchymal stem cells from rabbits[J].J Third Mil Med Univ,2008,30(16):275.

更新日期/Last Update: 2015-08-19