AIM: To evaluate the therapeutic effects of bone marrow-derived mesenchymal stem cells(BMSCs) with human urokinase-type plasminogen activator(u PA) on liver fibrosis, and to investigate the mechanism of gene therapy.M...AIM: To evaluate the therapeutic effects of bone marrow-derived mesenchymal stem cells(BMSCs) with human urokinase-type plasminogen activator(u PA) on liver fibrosis, and to investigate the mechanism of gene therapy.METHODS: BMSCs transfected with adenovirusmediated human urokinase plasminogen activator(Adu PA) were transplanted into rats with CCl4-induced liver fibrosis. All rats were sacrificed after 8 wk, and their serum and liver tissue were collected for biochemical, histopathologic, and molecular analyzes. The degree of liver fibrosis was assessed by hematoxylin and eosin or Masson's staining. Western blot and quantitative reverse transcription-polymerase chain reaction were used to determine protein and m RNA expression levels.RESULTS: Serum levels of alanine aminotransferase, aminotransferase, total bilirubin, hyaluronic acid, laminin, and procollagen type Ⅲ were markedly decreased, whereas the levels of serum albumin were increased by u PA gene modified BMSCs treatment. Histopathology revealed that chronic CCl4-treatment resulted in significant fibrosis while u PA gene modified BMSCs treatment significantly reversed fibrosis. By quantitatively analysing the fibrosis area of liver tissue using Masson staining in different groups of animals, we found that model animals with CCl4-induced liver fibrosis had the largest fibrotic area(16.69% ± 1.30%), while fibrotic area was significantly decreased by BMSCs treatment(12.38% ± 2.27%) and was further reduced by u PA-BMSCs treatment(8.31% ± 1.21%). Both protein and m RNA expression of β-catenin, Wnt4 and Wnt5 a was down-regulated in liver tissues following u PA gene modified BMSCs treatment when compared with the model animals.CONCLUSION: Transplantation of u PA gene modified BMSCs suppressed liver fibrosis and ameliorated liver function and may be a new approach to treating liver fibrosis. Furthermore, treatment with u PA gene modified BMSCs also resulted in a decrease in expression of molecules of the Wnt signaling pathway.展开更多
Background Patients with severe full-thickness burn injury suffer from their inability to maintain body temperature through perspiration because the complete destructed sweat glands can not be regenerated. Bone marrow...Background Patients with severe full-thickness burn injury suffer from their inability to maintain body temperature through perspiration because the complete destructed sweat glands can not be regenerated. Bone marrow-derived mesenchymal stem cells （BM-MSCs） represent an ideal stem-cell source for cell therapy because of their easy purification and multipotency. In this study, we attempted to induce human BM-MSCs to differentiate into sweat gland cells for sweat gland regeneration through ectodysplasin （EDA） gene transfection. Methods The dynamic expression of EDA and EDA receptor （EDAR） were firstly observed in the sweat gland formation during embryological development. After transfection with EDA expression vector, human BM-MSCs were transplanted into the injured areas of burn animal models. The regeneration of sweat glands was identified by perspiration test and immunohistochemical analysis. Results Endogenous expression of EDA and EDAR correlated with sweat gland development in human fetal skin. After EDA transfection, BM-MSC acquired a sweat-gland-cell phenotype, evidenced by their expression of sweat gland markers by flow cytometry analysis. Immunohistochemical staining revealed a markedly contribution of EDA-transfected BM-MSCs to the regeneration of sweat glands in the scalded paws. Positive rate for perspiration test for the paws treated with EDA-transfected BM-MSCs was significantly higher than those treated with BM-MSCs or EDA expression vector （P 〈0.05）. Conclusions Our results confirmed the important role of EDA in the development of sweat gland. BM-MSCs transfected with EDA significantly improved the sweat-gland regeneration. This study suggests the potential application of EDA-modified MSCs for the repair and regeneration of injured skin and its appendages.展开更多
Bone marrow-derived mesenchymal stem cells differentiate into neurons under the induction of Schwann cells. However, key microRNAs and related pathways for differentiation remain unclear. This study screened and ident...Bone marrow-derived mesenchymal stem cells differentiate into neurons under the induction of Schwann cells. However, key microRNAs and related pathways for differentiation remain unclear. This study screened and identified differentially expressed microRNAs in bone marrow- derived mesenchymal stem cells induced by Schwann cell-conditioned medium, and explored targets and related pathways involved in their differentiation into neuronal-like cells. Primary bone marrow-derived mesenchymal stem cells were isolated from femoral and tibial bones, while primary Schwann cells were isolated from bilateral saphenous nerves. Bone marrow-derived mesenchymal stem cells were cultured in unconditioned (control group) and Schwann cell-conditioned medium (bone marrow-derived mesenchymal stem cell + Schwann cell group). Neuronal differentiation of bone marrow-derived mesenchymal stem cells induced by Schwann cell-conditioned medium was observed by time-lapse imaging. Upon induction, the morphology of bone marrow-derived mesencaymal stem cells changed into a neural shape with neurites. Results of quantitative reverse transcription-polymerase chain reaction revealed that nestin mRNA expression was upregulated from 1 to 3 days and downregulated from 3 to 7 days in the bone marrow-derived mesenchymal stem cell + Schwann cell group. Compared with the control group, microtubule-associated protein 2 mRNA expression gradually increased from 1 to 7 days in the bone marrow-derived mesenchymal stem cell + Schwann cell group. After 7 days of induction, microRNA analysis iden:ified 83 significantly differentially expressed microRNAs between the two groups. Gene Ontology analysis indicated enrichment of microRNA target genes for neuronal projection development, regulation of axonogenesis, and positive regulation of cell proliferation. Kyoto Encyclopedia of Genes and Genomes pathway analysis demonstrated that Hippo, Wnt, transforming growth factor-beta, and Hedgehog signaling pathv/ays were potentially associated with neural differentiation of b展开更多
目的比较骨髓间充质干细胞(BMMSCs)经静脉和经气管2种移植方式对染矽尘大鼠的肺损伤修复作用。方法取14只无特定病原体级健康雄性SD大鼠培养BMMSCs,另取48只同类大鼠随机分为对照组、染尘组、静脉组和气管组,每组12只。对照组大鼠气管...目的比较骨髓间充质干细胞(BMMSCs)经静脉和经气管2种移植方式对染矽尘大鼠的肺损伤修复作用。方法取14只无特定病原体级健康雄性SD大鼠培养BMMSCs,另取48只同类大鼠随机分为对照组、染尘组、静脉组和气管组,每组12只。对照组大鼠气管内注入无菌生理氯化钠溶液1.0 m L,另3组大鼠均气管内注入质量浓度为40 g/L的无菌矽尘混悬液1.0 m L,静脉组、气管组再分别经鼠尾静脉和气管内注入细胞密度为5×109个/L的BMMSCs 0.5 m L。第28天放血处死大鼠,检测肺脏脏器系数;观察肺组织病理学改变情况,进行肺泡炎症和肺间质纤维化评分;检测肺组织羟脯氨酸(HYP)、转化生长因子-β1(TGF-β1)和肿瘤坏死因子-α(TNF-α)水平,以及周围血血常规和血生化指标。结果染尘组、静脉组和气管组大鼠肺脏脏器系数均高于对照组(P<0.05),静脉组和气管组大鼠肺脏脏器系数均低于染尘组(P<0.05),但静脉组和气管组大鼠肺脏脏器系数差异无统计学意义(P>0.05)。肺组织病理学观察显示:对照组大鼠肺组织结构正常;染尘组肺部炎症最严重,并可见大量矽结节和胶原纤维沉积;静脉组和气管组炎症、矽结节数量以及胶原纤维增生均较染尘组有改善。染尘组、静脉组和气管组大鼠的肺泡炎、肺间质纤维化评分以及肺组织TGF-β1、TNF-α水平均高于对照组(P<0.05),静脉组和气管组大鼠上述4个指标均低于染尘组(P<0.05),但静脉组和气管组大鼠上述4个指标差异均无统计学意义(P>0.05)。染尘组大鼠肺组织HYP水平高于其余3组(P<0.05);但静脉组、气管组和对照组HYP水平分别两两比较差异均无统计学意义(P>0.05)。染尘组、静脉组和气管组大鼠周围血白细胞计数、红细胞计数、血红蛋白水平、红细胞压积均分别高于对照组(P<0.05)。气管组大鼠血清尿素氮水平高于静脉组(P<0.05),染尘组大鼠血清铜蓝蛋白水平高于其余3组(P<0.展开更多
Sodium nitrite（Na NO2） is an inorganic salt used broadly in chemical industry. Na NO2 is highly reactive with hemoglobin causing hypoxia. Mesenchymal stem cells（MSCs） are capable of differentiating into a variety ...Sodium nitrite（Na NO2） is an inorganic salt used broadly in chemical industry. Na NO2 is highly reactive with hemoglobin causing hypoxia. Mesenchymal stem cells（MSCs） are capable of differentiating into a variety of tissue specific cells and MSC therapy is a potential method for improving brain functions. This work aims to investigate the possible therapeutic role of bone marrow-derived MSCs against Na NO2 induced hypoxic brain injury. Rats were divided into control group（treated for 3 or 6 weeks）, hypoxic（HP） group（subcutaneous injection of 35 mg/kg Na NO2 for 3 weeks to induce hypoxic brain injury）, HP recovery groups N-2 w R and N-3 w R（treated with the same dose of Na NO2 for 2 and 3 weeks respectively, followed by 4-week or 3-week self-recovery respectively）, and MSCs treated groups N-2 w SC and N-3 w SC（treated with the same dose of Na NO2 for 2 and 3 weeks respectively, followed by one injection of 2 × 106 MSCs via the tail vein in combination with 4 week self-recovery or intravenous injection of Na NO2 for 1 week in combination with 3 week self-recovery）. The levels of neurotransmitters（norepinephrine, dopamine, serotonin）, energy substances（adenosine monophosphate, adenosine diphosphate, adenosine triphosphate）, and oxidative stress markers（malondialdehyde, nitric oxide, 8-hydroxy-2′-deoxyguanosine, glutathione reduced form, and oxidized glutathione） in the frontal cortex and midbrain were measured using high performance liquid chromatography. At the same time, hematoxylin-eosin staining was performed to observe the pathological change of the injured brain tissue. Compared with HP group, pathological change of brain tissue was milder, the levels of malondialdehyde, nitric oxide, oxidized glutathione, 8-hydroxy-2′-deoxyguanosine, norepinephrine, serotonin, glutathione reduced form, and adenosine triphosphate in the frontal cortex and midbrain were significantly decreased, and glutathione reduced form/oxidized glutathione and adenosine monophosphate/adenosine tr展开更多
基金Supported by National Natural Science Foundation of ChinaNo.81460114+5 种基金Natural Science Foundation of Guangxi Zhuang Autonomous RegionNo.1355005-3-2 and No.2012GXNSFAA053143Chinese Traditional Medicine Science Foundation of Guangxi Zhuang Autonomous RegionNo.GZPT1238Science Foundation of Guangxi Department of EducationNo.201203YB036 and No.2013LX031
文摘AIM: To evaluate the therapeutic effects of bone marrow-derived mesenchymal stem cells(BMSCs) with human urokinase-type plasminogen activator(u PA) on liver fibrosis, and to investigate the mechanism of gene therapy.METHODS: BMSCs transfected with adenovirusmediated human urokinase plasminogen activator(Adu PA) were transplanted into rats with CCl4-induced liver fibrosis. All rats were sacrificed after 8 wk, and their serum and liver tissue were collected for biochemical, histopathologic, and molecular analyzes. The degree of liver fibrosis was assessed by hematoxylin and eosin or Masson's staining. Western blot and quantitative reverse transcription-polymerase chain reaction were used to determine protein and m RNA expression levels.RESULTS: Serum levels of alanine aminotransferase, aminotransferase, total bilirubin, hyaluronic acid, laminin, and procollagen type Ⅲ were markedly decreased, whereas the levels of serum albumin were increased by u PA gene modified BMSCs treatment. Histopathology revealed that chronic CCl4-treatment resulted in significant fibrosis while u PA gene modified BMSCs treatment significantly reversed fibrosis. By quantitatively analysing the fibrosis area of liver tissue using Masson staining in different groups of animals, we found that model animals with CCl4-induced liver fibrosis had the largest fibrotic area(16.69% ± 1.30%), while fibrotic area was significantly decreased by BMSCs treatment(12.38% ± 2.27%) and was further reduced by u PA-BMSCs treatment(8.31% ± 1.21%). Both protein and m RNA expression of β-catenin, Wnt4 and Wnt5 a was down-regulated in liver tissues following u PA gene modified BMSCs treatment when compared with the model animals.CONCLUSION: Transplantation of u PA gene modified BMSCs suppressed liver fibrosis and ameliorated liver function and may be a new approach to treating liver fibrosis. Furthermore, treatment with u PA gene modified BMSCs also resulted in a decrease in expression of molecules of the Wnt signaling pathway.
基金This work was supported by grants from the National Basic Science and Development Program of China （973 Program, No. 2005CB522603） and National Natural Science Foundation of China （No. 81000011, 81000835）, Distinguished Young Talents in Higher Education of Guangdong （No. LYM091182009）, and Shenzhen Technological R＆D Foundation （No. JC201005280429A）.
文摘Background Patients with severe full-thickness burn injury suffer from their inability to maintain body temperature through perspiration because the complete destructed sweat glands can not be regenerated. Bone marrow-derived mesenchymal stem cells （BM-MSCs） represent an ideal stem-cell source for cell therapy because of their easy purification and multipotency. In this study, we attempted to induce human BM-MSCs to differentiate into sweat gland cells for sweat gland regeneration through ectodysplasin （EDA） gene transfection. Methods The dynamic expression of EDA and EDA receptor （EDAR） were firstly observed in the sweat gland formation during embryological development. After transfection with EDA expression vector, human BM-MSCs were transplanted into the injured areas of burn animal models. The regeneration of sweat glands was identified by perspiration test and immunohistochemical analysis. Results Endogenous expression of EDA and EDAR correlated with sweat gland development in human fetal skin. After EDA transfection, BM-MSC acquired a sweat-gland-cell phenotype, evidenced by their expression of sweat gland markers by flow cytometry analysis. Immunohistochemical staining revealed a markedly contribution of EDA-transfected BM-MSCs to the regeneration of sweat glands in the scalded paws. Positive rate for perspiration test for the paws treated with EDA-transfected BM-MSCs was significantly higher than those treated with BM-MSCs or EDA expression vector （P 〈0.05）. Conclusions Our results confirmed the important role of EDA in the development of sweat gland. BM-MSCs transfected with EDA significantly improved the sweat-gland regeneration. This study suggests the potential application of EDA-modified MSCs for the repair and regeneration of injured skin and its appendages.
文摘目的 初步了解骨髓间充质干细胞（MSC）移植对烟雾吸入性损伤兔外周血主要炎症因子和肺水质量分数以及肺组织损伤的影响. 方法 取16只成年新西兰大耳白兔制成烟雾吸入性损伤模型后,按随机数字表法分成单纯致伤组和MSC移植组（各8只）.单纯致伤组伤后立即经耳缘静脉注入10 mL PBS;MSC移植组伤后立即经耳缘静脉注入10 mL PBS,内含兔第3代MSC（分离自健康幼龄新西兰大耳白兔）1×107个.另取8只成年新西兰大耳白兔作为正常对照组,不致伤,仅经耳缘静脉注入10 mL PBS.单纯致伤组和MSC移植组分别于致伤后2、4、6 h采血,以ELISA法检测血清TNF-α、IL-1β、IL-6、IL-10含量;伤后24 h,取兔肺行大体观察和组织病理学观察（左下肺）,取右肺中叶组织计算肺水质量分数.正常对照组同法抽血并取肺组织进行检测.对实验数据行t检验.结果 （1）两致伤组家兔各时相点血清TNF-α含量均明显高于正常对照组（t=2.43～9.57,P＜0.05或P＜0.01）.单纯致伤组各时相点血清IL-1β和IL-6含量明显高于正常对照组（t=8.49～19.80,P值均小于0.01）;MSC移植组各时相点IL-1β含量与正常对照组接近（t=0.11～0.92,P值均大于0.05）,IL-6含量伤后2 h与正常对照组接近（t=2.12,P＞0.05）,4、6 h显著升高（t值均为2.83,P值均小于0.05）.MSC移植组家兔各时相点血清TNF-α、IL-1β、IL-6明显低于单纯致伤组（t=2.35～12.45,P＜0.05或P＜0.01）.（2）MSC移植组伤后2、4、6 h血清IL-10含量分别为（13.0±3.6）、（11.6±8.5）、（15.2±4.4）pg/mL,与单纯致伤组各对应时相点的含量[分别为（5.5±3.4）、（5.0±1.7）、（7.9±3.5）pg/mL]相比均显著升高（t值分别为4.28、2.15、3.67,P值均小于0.01）.两致伤组亦明显高于正常对照组（t=2.46～8.14,P＜0.05或P＜0.01）.（3）大体观察及组织病理学观察见,MSC移植组肺组织损伤程度较单纯致伤组明显改善.（4）伤后24 h
基金supported by the National Natural Science Foundation of China,No.81330042,81620108018(both to SQF),and 81702147(to ZJW)
文摘Bone marrow-derived mesenchymal stem cells differentiate into neurons under the induction of Schwann cells. However, key microRNAs and related pathways for differentiation remain unclear. This study screened and identified differentially expressed microRNAs in bone marrow- derived mesenchymal stem cells induced by Schwann cell-conditioned medium, and explored targets and related pathways involved in their differentiation into neuronal-like cells. Primary bone marrow-derived mesenchymal stem cells were isolated from femoral and tibial bones, while primary Schwann cells were isolated from bilateral saphenous nerves. Bone marrow-derived mesenchymal stem cells were cultured in unconditioned (control group) and Schwann cell-conditioned medium (bone marrow-derived mesenchymal stem cell + Schwann cell group). Neuronal differentiation of bone marrow-derived mesenchymal stem cells induced by Schwann cell-conditioned medium was observed by time-lapse imaging. Upon induction, the morphology of bone marrow-derived mesencaymal stem cells changed into a neural shape with neurites. Results of quantitative reverse transcription-polymerase chain reaction revealed that nestin mRNA expression was upregulated from 1 to 3 days and downregulated from 3 to 7 days in the bone marrow-derived mesenchymal stem cell + Schwann cell group. Compared with the control group, microtubule-associated protein 2 mRNA expression gradually increased from 1 to 7 days in the bone marrow-derived mesenchymal stem cell + Schwann cell group. After 7 days of induction, microRNA analysis iden:ified 83 significantly differentially expressed microRNAs between the two groups. Gene Ontology analysis indicated enrichment of microRNA target genes for neuronal projection development, regulation of axonogenesis, and positive regulation of cell proliferation. Kyoto Encyclopedia of Genes and Genomes pathway analysis demonstrated that Hippo, Wnt, transforming growth factor-beta, and Hedgehog signaling pathv/ays were potentially associated with neural differentiation of b
文摘目的比较骨髓间充质干细胞(BMMSCs)经静脉和经气管2种移植方式对染矽尘大鼠的肺损伤修复作用。方法取14只无特定病原体级健康雄性SD大鼠培养BMMSCs,另取48只同类大鼠随机分为对照组、染尘组、静脉组和气管组,每组12只。对照组大鼠气管内注入无菌生理氯化钠溶液1.0 m L,另3组大鼠均气管内注入质量浓度为40 g/L的无菌矽尘混悬液1.0 m L,静脉组、气管组再分别经鼠尾静脉和气管内注入细胞密度为5×109个/L的BMMSCs 0.5 m L。第28天放血处死大鼠,检测肺脏脏器系数;观察肺组织病理学改变情况,进行肺泡炎症和肺间质纤维化评分;检测肺组织羟脯氨酸(HYP)、转化生长因子-β1(TGF-β1)和肿瘤坏死因子-α(TNF-α)水平,以及周围血血常规和血生化指标。结果染尘组、静脉组和气管组大鼠肺脏脏器系数均高于对照组(P<0.05),静脉组和气管组大鼠肺脏脏器系数均低于染尘组(P<0.05),但静脉组和气管组大鼠肺脏脏器系数差异无统计学意义(P>0.05)。肺组织病理学观察显示:对照组大鼠肺组织结构正常;染尘组肺部炎症最严重,并可见大量矽结节和胶原纤维沉积;静脉组和气管组炎症、矽结节数量以及胶原纤维增生均较染尘组有改善。染尘组、静脉组和气管组大鼠的肺泡炎、肺间质纤维化评分以及肺组织TGF-β1、TNF-α水平均高于对照组(P<0.05),静脉组和气管组大鼠上述4个指标均低于染尘组(P<0.05),但静脉组和气管组大鼠上述4个指标差异均无统计学意义(P>0.05)。染尘组大鼠肺组织HYP水平高于其余3组(P<0.05);但静脉组、气管组和对照组HYP水平分别两两比较差异均无统计学意义(P>0.05)。染尘组、静脉组和气管组大鼠周围血白细胞计数、红细胞计数、血红蛋白水平、红细胞压积均分别高于对照组(P<0.05)。气管组大鼠血清尿素氮水平高于静脉组(P<0.05),染尘组大鼠血清铜蓝蛋白水平高于其余3组(P<0.
文摘Sodium nitrite（Na NO2） is an inorganic salt used broadly in chemical industry. Na NO2 is highly reactive with hemoglobin causing hypoxia. Mesenchymal stem cells（MSCs） are capable of differentiating into a variety of tissue specific cells and MSC therapy is a potential method for improving brain functions. This work aims to investigate the possible therapeutic role of bone marrow-derived MSCs against Na NO2 induced hypoxic brain injury. Rats were divided into control group（treated for 3 or 6 weeks）, hypoxic（HP） group（subcutaneous injection of 35 mg/kg Na NO2 for 3 weeks to induce hypoxic brain injury）, HP recovery groups N-2 w R and N-3 w R（treated with the same dose of Na NO2 for 2 and 3 weeks respectively, followed by 4-week or 3-week self-recovery respectively）, and MSCs treated groups N-2 w SC and N-3 w SC（treated with the same dose of Na NO2 for 2 and 3 weeks respectively, followed by one injection of 2 × 106 MSCs via the tail vein in combination with 4 week self-recovery or intravenous injection of Na NO2 for 1 week in combination with 3 week self-recovery）. The levels of neurotransmitters（norepinephrine, dopamine, serotonin）, energy substances（adenosine monophosphate, adenosine diphosphate, adenosine triphosphate）, and oxidative stress markers（malondialdehyde, nitric oxide, 8-hydroxy-2′-deoxyguanosine, glutathione reduced form, and oxidized glutathione） in the frontal cortex and midbrain were measured using high performance liquid chromatography. At the same time, hematoxylin-eosin staining was performed to observe the pathological change of the injured brain tissue. Compared with HP group, pathological change of brain tissue was milder, the levels of malondialdehyde, nitric oxide, oxidized glutathione, 8-hydroxy-2′-deoxyguanosine, norepinephrine, serotonin, glutathione reduced form, and adenosine triphosphate in the frontal cortex and midbrain were significantly decreased, and glutathione reduced form/oxidized glutathione and adenosine monophosphate/adenosine tr
