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bone marrow mesenchymal stem cells and exercise restore motor function following spinal cord injury by activating PI3K/AKT/mTOR pathway

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Neural Regeneration Research 2023年第5期18卷 1067-1075页

作者： Xin Sun Li-Yi Huang Hong-Xia Pan Li-Juan Li Lu Wang Gai-Qin Pei Yang Wang Qing Zhang Hong-Xin Cheng Cheng-Qi He Quan Wei Rehabilitation Medical Center and Institute of Rehabilitation Medicine West China HospitalSichuan UniversityChengduSichuan ProvinceChina Key Laboratory of Rehabilitation Medicine in Sichuan Province ChengduSichuan ProvinceChina

Although many therapeutic interventions have shown promise in treating spinal cord injury, focusing on a single aspect of repair cannot achieve successful and functional regeneration in patients following spinal cord injury. In this study, we applied a combinatorial approach for treating spinal cord injury involving neuroprotection and rehabilitation, exploiting cell transplantation and functional sensorimotor training to promote nerve regeneration and functional recovery. Here, we used a mouse model of thoracic contusive spinal cord injury to investigate whether the combination of bone marrow mesenchymal stem cell transplantation and exercise training has a synergistic effect on functional restoration. Locomotor function was evaluated by the Basso Mouse Scale, horizontal ladder test, and footprint analysis. Magnetic resonance imaging, histological examination, transmission electron microscopy observation, immunofluorescence staining, and western blotting were performed 8 weeks after spinal cord injury to further explore the potential mechanism behind the synergistic repair effect. In vivo, the combination of bone marrow mesenchymal stem cell transplantation and exercise showed a better therapeutic effect on motor function than the single treatments. Further investigations revealed that the combination of bone marrow mesenchymal stem cell transplantation and exercise markedly reduced fibrotic scar tissue, protected neurons, and promoted axon and myelin protection. Additionally, the synergistic effects of bone marrow mesenchymal stem cell transplantation and exercise on spinal cord injury recovery occurred via the PI3 K/AKT/mTOR pathway. In vitro, experimental evidence from the PC12 cell line and primary cortical neuron culture also demonstrated that blocking of the PI3 K/AKT/mTOR pathway would aggravate neuronal damage. Thus, bone marrow mesenchymal stem cell transplantation combined with exercise training can effectively restore motor function after spinal cord injury by activating the PI3 K/AKT/mTOR pathway.

关键词： axon growth bone marrow mesenchymal stem cell exercise training mTOR neuroprotection neurotrophin remyelination scar formation spinal cord injury synaptic plasticity

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Overexpression of vascular endothelial growth factor enhances the neuroprotective effects of bone marrow mesenchymal stem cell transplantation in ischemic stroke

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Neural Regeneration Research 2023年第6期18卷 1286-1292页

作者： Cui Liu Zhi-Xiang Yang Si-Qi Zhou Ding Ding Yu-Ting Hu Hong-Ning Yang Dong Han Shu-Qun Hu Xue-Mei Zong Institute of Emergency Rescue Medicine Emergency Centerthe Affiliated Hospital of Xuzhou Medical UniversityXuzhouJiangsu ProvinceChina Emergency Department of Affiliated Lianyungang Hospital of Xuzhou Medical University/The First People’s Hospital of Lianyungang LianyungangLianyungangJiangsu ProvinceChina Laboratory of Emergency Medicine Second Clinical Medical College of Xuzhou Medical UniversityXuzhou Medical UniversityXuzhouJiangsu ProvinceChina Key Laboratory of Brain Diseases Bioinformation Xuzhou Medical UniversityXuzhouJiangsu ProvinceChina

Although bone marrow mesenchymal stem cells(BMSCs)might have therapeutic potency in ischemic stroke,the benefits are limited.The current study investigated the effects of BMSCs engineered to overexpress vascular endothelial growth factor(VEGF)on behavioral defects in a rat model of transient cerebral ischemia,which was induced by middle cerebral artery occlusion.VEGF-BMSCs or control grafts were injected into the left striatum of the infarcted hemisphere 24 hours after stroke.We found that compared with the stroke-only group and the vehicle-and BMSCs-control groups,the VEGF-BMSCs treated animals displayed the largest benefits,as evidenced by attenuated behavioral defects and smaller infarct volume 7 days after stroke.Additionally,VEGF-BMSCs greatly inhibited destruction of the blood-brain barrier,increased the regeneration of blood vessels in the region of ischemic penumbra,and reducedneuronal degeneration surrounding the infarct core.Further mechanistic studies showed that among all transplant groups,VEGF-BMSCs transplantation induced the highest level of brain-derived neurotrophic factor.These results suggest that BMSCs transplantation with vascular endothelial growth factor has the potential to treat ischemic stroke with better results than are currently available.

关键词： bone marrow mesenchymal stem cell brain-derived neurotrophic factor CD31 microtubule associated protein 2 middle cerebral artery occlusion stroke transplantation vascular endothelial growth factor

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Combination of low-energy shock-wave therapy and bone marrow mesenchymal stem cell transplantation to improve the erectile function of diabetic rats

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Asian Journal of Andrology 2017年第1期19卷 26-33页

作者： Hai-Tao Shan Hai-Bo Zhang Wen-Tao Chen Feng-Zhi Chen Tao Wang Jin-Tai Luo Min Yue Ji-Hong Lin An-Yang Wei Department of Urology Medical Center for Overseas Patients Nanfang Hospital Southern Medical University Guangzhou China Department of Urology Shawan People's Hospital Panyu District Guangzhou China Shenzhen Hyde Medical Equipment Co. Ltd. Shenzhen China Department of Urology Longjiang HospitalShunde District Foshan China Laboratory Animals Center Southern Medical University Guangzhou China.

stem cell transplantation and low-energy shock-wave therapy （LESWT） have emerged as potential and effective treatment protocols for diabetic erectile dysfunction. During the tracking of transplanted stem cells in diabetic erectile dysfunction models, the number of visible stem cells was rather low and decreased quickly. LESWT could recruit endogenous stem cells to the cavernous body and improve the microenvironment in diabetic cavernous tissue. Thus, we deduced that LESWT might benefit transplanted stem cell survival and improve the effects of stem cell transplantation. In this research, 42 streptozotocin-induced diabetic rats were randomized into four groups： the diabetic group （n = 6）, the LESWT group （n = 6）, the bone marrow-derived mesenchymal stem cell （BMSC） transplantation group （n = 15）, and the combination of LESWT and BMSC transplantation group （n = 15）. One and three days after BMSC transplantation, three rats were randomly chosen to observe the survival numbers of BMSCs in the cavernous body. Four weeks after BMSC transplantation, the following parameters were assessed： the surviving number of transplanted BMSCs in the cavernous tissue, erectile function, real-time polymerase chain reaction, and penile immunohistochemical assessment. Our research found that LESWT favored the survival of transplanted BMSCs in the cavernous body, which might be related to increased stromal cell-derived factor-1 expression and the enhancement of angiogenesis in the diabetic cavernous tissue. The combination of LESWT and BMSC transplantation could improve the erectile function of diabetic erectile function rats more effectively than LESWT or BMSC transplantation performed alone.

关键词： bone marrow mesenchymal stem cell diabetes mellitus erectile function low-energy shock-wave therapy

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bone marrow mesenchymal stem cells induce M2 microglia polarization through PDGF-AA/MANF signaling

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World Journal of stem cells 2020年第7期12卷 633-657页

作者： Fan Yang Wen-Bin Li Ye-Wei Qu Jin-Xing Gao Yu-Shi Tang Dong-Jie Wang Yu-Jun Pan Department of Neurology The First Clinical College of Harbin Medical UniversityHarbin 150001Heilongjiang ProvinceChina Department of Respiratory Medicine The First Clinical College of Harbin Medical UniversityHarbin 150001Heilongjiang ProvinceChina

BACKGROUND bone marrow mesenchymal stem cells(BMSCs)are capable of shifting the microglia/macrophages phenotype from M1 to M2,contributing to BMSCsinduced brain repair.However,the regulatory mechanism of BMSCs on microglia/macrophages after ischemic stroke is unclear.Recent evidence suggests that mesencephalic astrocyte-derived neurotrophic factor(MANF)and plateletderived growth factor-AA(PDGF-AA)/MANF signaling regulate M1/M2 macrophage polarization.AIM To investigate whether and how MANF or PDGF-AA/MANF signaling influences BMSCs-mediated M2 polarization.METHODS We identified the secretion of MANF by BMSCs and developed transgenic BMSCs using a targeting small interfering RNA for knockdown of MANF expression.Using a rat middle cerebral artery occlusion(MCAO)model transplanted by BMSCs and BMSCs-microglia Transwell coculture system,the effect of BMSCsinduced downregulation of MANF expression on the phenotype of microglia/macrophages was tested by Western blot,quantitative reverse transcription-polymerase chain reaction,and immunofluorescence.Additionally,microglia were transfected with mimics of miR-30a*,which inuenced expression of X-box binding protein(XBP)1,a key transcription factor that synergized with activating transcription factor 6(ATF6)to govern MANF expression.We examined the levels of miR-30a*,ATF6,XBP1,and MANF after PDGF-AA treatment in the activated microglia.RESULTS Inhibition of MANF attenuated BMSCs-induced functional recovery and decreased M2 marker production,but increased M1 marker expression in vivo or in vitro.Furthermore,PDGF-AA treatment decreased miR-30a*expression,had no influence on the levels of ATF6,but enhanced expression of both XBP1 and MANF.CONCLUSION BMSCs-mediated MANF paracrine signaling,in particular the PDGF-AA/miR-30a*/XBP1/MANF pathway,synergistically mediates BMSCs-induced M2 polarization.

关键词： Mesencephalic astrocyte-derived neurotrophic factor bone marrow mesenchymal stem cell Microglia/macrophage polarization Endoplasmic reticulum stress Cerebral ischemia/reperfusion injury

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Baicalin attenuates dexamethasone-induced apoptosis of bone marrow mesenchymal stem cells by activating the hedgehog signaling pathway

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Chinese Medical Journal 2023年第15期136卷 1839-1847页

作者： Bin Jia Yaping Jiang Yao Yao Yingxing Xu Yingzhen Wang Tao Li Department of Joint Surgery The Affiliated Hospital of Qingdao UniversityQingdaoShandong 266003China Medical Department Qingdao UniversityQingdaoShandong 266071China Department of Oral Implantology The Affiliated Hospital of Qingdao UniversityQingdaoShandong 266003China

Background:Perturbations in bone marrow mesenchymal stem cell(BMSC)differentiation play an important role in steroid-induced osteonecrosis of the femoral head(SONFH).At present,studies on SONFH concentrate upon the balance within BMSC osteogenic and adipogenic differentiation.However,BMSC apoptosis as well as proliferation are important prerequisites in their differentiation.The hedgehog(HH)signaling pathway regulates bone cell apoptosis.Baicalin(BA),a well-known compound in traditional Chinese medicine,can affect the proliferation and apoptosis of numerous cell types via HH signaling.However,the potential role and mechanisms of BA on BMSCs are unclear.Thus,we aimed to explore the role of BA in dexamethasone(Dex)-induced BMSC apoptosis in this study.Methods:Primary BMSCs were treated with 10-6 mol/L Dex alone or with 5.0μmol/L,10.0μmol/L,or 50.0μmol/L BA for 24 hours followed by co-treatment with 5.0μmol/L,10.0μmol/L,or 50.0μmol/L BA and 10-6 mol/L Dex.cell viability was assayed through the cell Counting Kit-8(CCK-8).cell apoptosis was evaluated using Annexin V-fluorescein isothiocyanate/propidium iodide(PI)staining followed by flow cytometry.The imaging and counting,respectively,of Hochest 33342/PI-stained cells were used to assess the morphological characteristics and proportion of apoptotic cells.To quantify the apoptosis-related proteins(e.g.,apoptosis regulator BAX[Bax],B-cell lymphoma 2[Bcl-2],caspase-3,and cleaved caspase-3)and HH signaling pathway proteins,western blotting was used.A HH-signaling pathway inhibitor was used to demonstrate that BA exerts its anti-apoptotic effects via the HH signaling pathway.Results:The results of CCK-8,Hoechst 33342/PI-staining,and flow cytometry showed that BA did not significantly promote cell proliferation(CCK-8:0μmol/L,100%;2.5μmol/L,98.58%;5.0μmol/L,95.18%;10.0μmol/L,98.11%;50.0μmol/L,99.38%,F=2.33,P>0.05),but it did attenuate the effect of Dex on apoptosis(Hoechst 33342/PI-staining:Dex+50.0μmol/L BA,12.27%vs.Dex,39.27%,t=20.62;flow cytometry:Dex+50.0μmol/L BA,12.68%

关键词： Baicalin Dexamethasone Apoptosis bone marrow mesenchymal stem cell Hedgehog

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Chinese preparation Xuesaitong promotes the mobilization of bone marrow mesenchymal stem cells in rats with cerebral infarction

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Neural Regeneration Research 2016年第2期11卷 292-297页

作者： Jin-sheng Zhang Bao-xia Zhang Mei-mei Du Xiao-ya Wang Wei Li First Affiliated Hospital of Henan University of Traditional Chinese Medicine Zhengzhou Henan Province China Third Affiliated Hospital of Henan University of Traditional Chinese Medicine Zhengzhou Henan Province China

After cerebral ischemia, bone marrow mesenchymal stem cells are mobilized and travel from the bone marrow through peripheral circulation to the focal point of ischemia to initiate tissue regeneration. However, the number of bone marrow mesenchymal stem cells mobilized into peripheral circulation is not enough to exert therapeutic effects, and the method by which blood circulation is promoted to remove blood stasis influences stem cell homing. The main ingredient of Xuesaitong capsules is Panax notoginseng saponins, and Xuesaitong is one of the main drugs used for promoting blood circulation and removing blood stasis. We established rat models of cerebral infarction by occlusion of the middle cerebral artery and then intragastrically administered Xuesaitong capsules（20, 40 and 60 mg/kg per day） for 28 successive days. Enzyme-linked immunosorbent assay showed that in rats with cerebral infarction, middle- and high-dose Xuesaitong significantly increased the level of stem cell factors and the number of CD117-positive cells in plasma and bone marrow and significantly decreased the number of CD54-and CD106-positive cells in plasma and bone marrow. The effect of low-dose Xuesaitong on these factors was not obvious. These findings demonstrate that middle- and high-dose Xuesaitong and hence Panax notoginseng saponins promote and increase the level and mobilization of bone marrow mesenchymal stem cells in peripheral blood.

关键词： nerve regeneration Panax notoginseng saponin Xuesaitong bone marrow mesenchymal stem cell cerebral infarction mobilization peripheral circulation homing nerual regeneration

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Characterization and Differentiation into Adipocytes and Myocytes of Porcine bone marrow mesenchymal stem cells

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Journal of Integrative Agriculture 2014年第4期13卷 837-848页

作者： DU Min-qing HUANG Yue-qin LU Nai-Sheng SHU Gang ZHU Xiao-tong WANG Li-na GAO Ping XI Qian-yun ZHANG Yong-liang WANG Song-bo JIANG Qing-yan College of Animal Sciences South China Agricultural University

bone marrow mesenchymal stem cells （BMSCs） could differentiate into various cell types including adipocytes and myocytes, which had important scientific significance not only in the field of tissue regeneration, but also in the field of agricultural science. In an attempt to exhibit the characterization and differentiation into adipocytes and myocytes of porcine BMSCs, we isolated and purified porcine BMSCs by red blood cell lysis method and percoll gradient centrifugation. The purified cells presented a stretched fibroblast-like phenotype when adhered to the culture plate. The results of flow cytometry analysis and immunofluorescence staining demonstrated that the isolated cells were positive for mesenchymal surface markers CD29, CD44 and negative for hematopoietic markers CD45 and the adhesion molecules CD31. cells were induced to differentiate into adipocytes with adipogenic medium containing insulin, dexamethasone, oleate and octanoate. Oil Red O staining demonstrated that the porcine BMSCs successfully differentiated to adipocytes. Moreover, the findings of real-time PCR and Western blotting indicated that the induced cells expressed adipogenic marker genes （PPAR-y, C/EBP-c~, perilipin, aP2） mRNA or proteins （PPAR-3,, perilipin, aP2）. On the other hand, porcine BMSCs were induced into myoctyes with myogenic medium supplemented with 5-azacytidine, basic fibroblast growth factor, chick embryo extract and horse serum. Morphological observation by hochest 33342 staining showed that the induced cells presented as multi-nucleus muscular tube structure. And myogenic marker genes （Myf5, desmin） mRNA or proteins （MyfS, MyoD, myogenin, desmin） were found in the induced cells. In addition, the results of immunofluorescence staining revealed that myogenic marker （Myf5, MyoD, myogenin, desmin, S-MyHC） proteins was positive in the induced cells. Above all, these results suggested that the isolated porcine BMSCs were not only consistent with the characterization of mesenchymal stem cells, but also exhibited the multipotent

关键词： porcine bone marrow mesenchymal stem cell adipocyte myocyte

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Chondrogenic Differentiation of Mouse bone marrow mesenchymal stem cells Induced by Cartilage-derived Morphogenetic Protein-2 In Vitro

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2007年第4期27卷 429-432页

作者：田洪涛杨述华徐亮张宇坤许伟华 Department of Orthopedics Union Hospital Tongji Medical College Huazhong University of Science and Technology

To study the cartilage differentiation of mouse mesenchymal stem cells （MSCs） induced by cartilage-derived morphogenetic proteins-2 in vitro, the MSCs were isolated from mouse bone marrow and cultured in vitro. The cells in passage 3 were induced into chondrogenic differentiation with different concentrations of recombinant human cartilage-derived morphogenetic proteins-2 （0, 10, 20, 50 and 100 ng/mL）. After 14 days of induction, morphology of cells was observed under phase-contrast microscope. Collagen Ⅱ mRNA and protein were examined with RT-PCR, Western blotting and immunocytochemistry respectively and the sulfate glycosaminoglycan was measured by Alcian blue staining. RT-PCR showed that CDMP-2 could promote expression of collagen Ⅱ mRNA in an dose-dependant manner, especially at the concentration of 50 ng/mL and 100 ng/mL. Immunocytochemistry and Western blotting revealed a similar change. Alcian blue staining exhibited deposition of typical cartilage extracellular matrix. Our results suggest that mouse bone marrow mesencymal stem cells can differentiate into chondrogenic phonotype with the induction of CDMP-2 in vitro, which provides a basis for further research on the role of CDMP-2 in chondrogenesis.

关键词： cartilage-derived morphogenetic proteins-2 bone marrow mesenchymal stem cell chondrogenic differentiation mouse

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Improvement of cardiac function after transplantation of autologous bone marrow mesenchymal stem cells in patients with acute myocardial infarction

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Chinese Medical Journal 2004年第10期117卷 1443-1448页

作者：陈绍良方五旺钱钧叶飞刘煜昊单守杰张俊杰林松廖联明赵春华 Department of Cardiology Nanjing First Municipal Hospital Nanjing Medical University Nanjing 210006 China Department of Cardiology Maanshan Municipal People’s Hospital Maanshan 240058 China Tissue and Engineering Centre Chinese Academy of Medical Sciences Beijing 100036 China

Background The infarct size determines the long-term prognosis of patients with acute myocardial infarction (AMI). There is a growing interest in repairing scar area by transplanting bone marrow stem cells. However, effectiveness of intracoronary injection of bone marrow mesenchymal stem cells (BMSCs) in patients with AMI still remains unclear.Methods Sixty-nine patients with AMI after percutaneous coronary intervention (PCI) were randomly divided into intracoronary injection of BMSCs (n=34) and saline (control group, n=35) groups. Serial single positron emission computer tomography (SPECT), cardiac echo and cardiac electromechanical mapping were done at the designed time intervals until six months after transplantation of BMSCs or injection of saline. Results The proportion with functional defect decreased significantly in the BMSCs patients after three months [(13±5)%] compared with that pre-transplantation [(32±11)%] and the control group [(28±10)%] at three month follow-up (P0.05]. Left ventricular ejection fraction (LVEF) three months after transplantation in BMSCs group increased significantly compared with that pre-implantation and with that of the control group at three months post-injection [(67±11)% vs (49±9)% and (53±8)%, P<0.05 respectively]. SPECT scan results showed that perfusion defect was improved significantly in BMSCs group at three-month follow-up compared with that in the control group [(134±66)cm2 vs (185±87)cm2, P<0.01]. At the same time, left ventricular end-diastolic volume [(136±31) ml vs (162±27) ml, P<0.05] and end-systolic volume [(63±20) ml vs (88±19) ml, P<0.05] decreased synchronously. The ratio of end-systolic pressure to end-systolic volume [P_ syst/ESV, (2.84±1.30) mmHg/ml vs (1.72±1.23) mmHg/ml, P<0.05] increased significantly. Cardiac electromechnical

关键词： acute myocardial infarction bone marrow mesenchymal stem cell cell transplantation

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Nystose attenuates bone loss and promotes BMSCs differentiation to osteoblasts through BMP and Wnt/β-catenin pathway in ovariectomized mice

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Food Science and Human Wellness 2023年第2期12卷 634-646页

作者： Qi Zhang Sijing Hu Jianjun Wu Peng Sun Quanlong Zhang Yang Wang Qiming Zhao Ting Han Luping Qin Qiaoyan Zhang School of Pharmaceutical Sciences Zhejiang Chinese Medical UniversityHangzhou 310053China Zhejiang Traditional Chinese Medicine&Health Industry Group Co. Ltd.Hangzhou 310016China School of Pharmacy Second Military Medical UniversityShanghai 200433China

Increasing the osteogenic differentiation ability and decreasing the adipogenic differentiation ability of bone marrow mesenchymal stem cells(BMSCs)is a potential strategy for the treatment of osteoporosis(OP).Naturally derived oligosaccharides have shown significant anti-osteoporotic effects.Nystose(NST),an oligosaccharide,was isolated from the roots of Morinda officinalis How.(MO).The aim of the present study was to investigate the effects of NST on bone loss in ovariectomized mice,and explore the underlying mechanism of NST in promoting differentiation of BMSCs to osteoblasts.Administration of NST(40,80 and 160 mg/kg)and the positive control of estradiol valerate(0.2 mg/kg)for 8 weeks significantly prevented bone loss induced by ovariectomy(OVX),increased the bone mass density(BMD),improved the bone microarchitecture and reduced urine calcium and deoxypyridinoline(DPD)in ovariectomized mice,while inhibited the increase of body weight without significantly affecting the uterus weight.Furthermore,we found that NST increased osteogenic differentiation,inhibited adipogenic differentiation of BMSCs in vitro,and upregulated the expression of the key proteins of BMP and Wnt/β-catenin pathways.In addition,Noggin and Dickkopf-related protein-1(DKK-1)reversed the effect of NST on osteogenic differentiation and expression of the key proteins in BMP and Wnt/β-catenin pathway.The luciferase activities and the molecular docking analysis further supported the mechanism of NST.In conclusion,these results indicating that NST can be clinically used as a potential alternative medicine for the prevention and treatment of postmenopausal osteoporosis.

关键词： Nystose bone marrow mesenchymal stem cell Osteoblast Adipocyte BMP pathway Wnt/β-catenin pathway

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