Therapeutic effect of bone marrow mesenchymal stem cells on multiple organ dysfunction syndrome

doi:10.3969/j.issn.2095-4344.2017.09.013

Chinese Journal of Tissue Engineering Research ›› 2017, Vol. 21 ›› Issue (9): 1378-1383.doi: 10.3969/j.issn.2095-4344.2017.09.013

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Therapeutic effect of bone marrow mesenchymal stem cells on multiple organ dysfunction syndrome

He Fa-lian^{1, 2}, Liu Gao-mi-yang¹, Liu Ju-fen¹, Li Zi-an¹, Wang Qiang¹, Pan Xing-hua¹

¹Cell Biological Therapy Center of Kunming General Hospital of Chengdu Military Area of Chinese PLA, Cell Biological Medicine Integrated Engineering Laboratory of State and Regions in Yunnan Province, the Stem Cell Therapy Key Laboratory of Yunnan Province, Kunming 650032, Yunnan Province, China; ²Kunming General Hospital of Chengdu Military Region Kunming Medical University Clinical College, Kunming 650031, Yunnan Province, China

Online:2017-03-28 Published:2017-03-31
Contact: Pan Xing-hua, M.D., Chief physician, Master’s and doctoral supervisor, Cell Biological Therapy Center of Kunming General Hospital of Chengdu Military Area of Chinese PLA, Cell Biological Medicine Integrated Engineering Laboratory of State and Regions in Yunnan Province, the Stem Cell Therapy Key Laboratory of Yunnan Province, Kunming 650032, Yunnan Province, China
About author:He Fa-lian, Studying for master’s degree, Cell Biological Therapy Center of Kunming General Hospital of Chengdu Military Area of Chinese PLA, Cell Biological Medicine Integrated Engineering Laboratory of State and Regions in Yunnan Province, the Stem Cell Therapy Key Laboratory of Yunnan Province, Kunming 650032, Yunnan Province, China; Kunming General Hospital of Chengdu Military Region Kunming Medical University Clinical College, Kunming 650031, Yunnan Province, China Liu Gao-mi-yang, Master, Cell Biological Therapy Center of Kunming General Hospital of Chengdu Military Area of Chinese PLA, Cell Biological Medicine Integrated Engineering Laboratory of State and Regions in Yunnan Province, the Stem Cell Therapy Key Laboratory of Yunnan Province, Kunming 650032, Yunnan Province, China He Fa-lian and Liu Gao-mi-yang contributed equally to this work.
Supported by:
the National Sci-Tech Support Plan of China, No. 2014BI01B01-05; the Major Scientific Plan of Yunnan Province, No. 2013CA005; the PLA Special Fund for Experimental Animals, No. DW2014006

Abstract

Abstract:

BACKGROUND: Clinical treatment of multiple organ dysfunction syndrome has been a big problem today, while the treatment of bone marrow mesenchymal stem cells may provide a new research direction.

OBJECTIVE: To investigate the therapeutic effects of bone marrow mesenchymal stem cells transplanted into multiple organ dysfunction syndrome models.

METHODS: According to body weight of rats, endotoxin in the 1 mg/kg dose was injected into the femoral vein to build the multiple organ dysfunction syndrome models in rats, and the surviving rats were randomly divided into treatment and control groups. Then the rats in the treatment group were injected intravenously third-generation bone marrow mesenchymal stem cells (5×10⁶), while rats in the control group injected intravenously with an equal volume of saline. After 5 days of treatment, the blood routine examination, blood biochemistry and coagulation tests were conducted, and then the liver, kidney, heart, and spleen of rats were captured to observed pathological changes through the hematoxylin-eosin staining.

RESULTS AND CONCLUSION: One week after cell transplantation, liver function and biochemical indicators were improved significantly in the treatment group compared with the control group (P < 0.05), and the white blood cell count in the treatment group was closer to normal levels (P < 0.05). Hematoxylin-eosin staining results showed that in the multiple organ dysfunction syndrome rats appeared to have different degrees of inflammatory lesions and necrosis in the heart, liver, spleen, and kidneys, while inflammatory infiltration and necrotic lesions decreased significantly in the treatment group compared with the control group. To conclude, bone marrow mesenchymal stem cells exert remarkable therapeutic effects on multiple organ dysfunction syndrome.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Endotoxins, Multiple Organ Failure, Bone Marrow, Mesenchymal Stem Cell Transplantation, Tissue Engineering

CLC Number:

R394.2

He Fa-lian, Liu Gao-mi-yang, Liu Ju-fen, Li Zi-an, Wang Qiang, Pan Xing-hua . Therapeutic effect of bone marrow mesenchymal stem cells on multiple organ dysfunction syndrome[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(9): 1378-1383.

Figures/Tables 2

2.1 骨髓间充质干细胞培养、扩增以及诱导鉴定结果采用贴壁培养法分离培养骨髓间充质干细胞，接种两三天后可见少量贴壁细胞生长，呈梭形或多边形，表面覆盖大量的杂细胞，随着培养时间延长，大量细胞贴壁，换液后可见形态均一的细胞增殖集落(图1A)；培养10-13 d，95%以上细胞融合。经胰酶消化后1︰2传代，细胞贴壁后呈典型的成纤维细胞形态(图1B)，分布均匀，大小一致，生长5-7 d后传代。流式细胞仪检测结果显示干细胞标记物CD29，CD44，CD90的阳性表达率为99.9%，96%和98.6%，干细胞阴性标记物CD34的表达率为6.94%，结果说明移植所用的细胞符合间充质干细胞的表型特征，见图2。对第3代骨髓间充质干细胞进行体外诱导分化实验，成脂诱导后，细胞变圆，形态不规则，出现脂肪小滴，经油红O染色后，脂滴明显红染、沉积。成骨诱导后经茜素红染色，出现明显红染的钙结节[4]，见图3。 2.2 骨髓间充质干细胞治疗前后生物化学检验相关指标变化肝功能结果显示，在细胞移植治疗5 d后，治疗组总蛋白、白蛋白水平均明显高于模型组(P < 0.05)，蛋白水平有恢复趋势，但仍未达到正常水平；谷丙转氨酶、谷草转氨酶较模型组有明显下降(P < 0.05)，且谷草转氨酶仍高于正常值3倍以上[3]，差异有显著性意义。心功能结果显示，治疗组乳酸脱氢酶、肌酸磷酸激酶、肌酸激酶同工酶均下降，其中乳酸脱氢酶由(2 191.50±31.27) U/L降至(1 197.00 ±36.38) U/L，与模型组相比差异有显著性意义(P < 0.05)，见表1。肾脏生化指标尿素氮、肌酐未发生明显变化。 2.3 骨髓间充质干细胞治疗前后凝血功能相关指标变化凝血功能显示，在细胞移植5 d后，治疗组凝血酶原时间、凝血酶时间较模型组缩短，差异有显著性意义(P < 0.05)，见表2。 2.4 骨髓间充质干细胞治疗前后炎症细胞分析结果造模后，白细胞计数增高1倍或减少50%，满足全身炎症反应条件中对白细胞的要求。经骨髓间充质干细胞治疗后，有助于白细胞趋于正常水平，差异均有显著性意义(P < 0.05)，见表3。 2.5 组织病理学观察结果 2.5.1 肝脏病理组织结构模型组苏木精-伊红染色显示肝小叶结构破坏，肝索排列紊乱，窦间隙变窄；肝细胞肿大，脂肪变性明显，核大而深染、核型不整齐；血管扩张充血明显，血窦充血，炎性细胞浸润明显，说明存在急性肝炎病变(图4A)；骨髓间充质干细胞治疗后，肝脏上述症状变轻，肝脏变性、坏死细胞明显减轻，炎症浸润减少，肝细胞围绕中央静脉呈放射状排列，可见肝脏病变形态趋于修复(图4B)。 2.5.2 心肌病理组织结构模型组苏木精-伊红染色显示心肌细胞脂肪变性，呈空泡状，与未受侵犯的红色心肌相间排列(图5A)；治疗组脂肪空泡较少，心肌排列整齐、紧密，与模型组相比，心肌修复明显(图5B)。 2.5.3 脾脏病理组织结构模型组苏木精-伊红染色显示脾脏充血淤血很明显，大量红细胞充满视野，淋巴细胞等被其覆盖、挤压，炎症反应明显(图6A)。与模型组相比，治疗组充血明显减轻，红细胞覆盖减少，大量淋巴细胞显现，多核细胞等炎症修复细胞出现，脾脏病变形态趋于修复(图6B)。 2.5.4 肾脏病理组织结构模型组苏木精-伊红染色显示肾小球肥大，基底膜增厚，肾小球囊腔增大；肾小管管壁增厚，有玻璃样物质沉积，毛细血管扩张充血，炎症细胞浸润(图7A)。与模型组相比，治疗组肾脏病变明显减轻，炎性细胞浸润明显减少，毛细血管充血减轻，肾小球囊腔趋于正常，可明显看到肾组织得到修复(图7B)。"

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Therapeutic effect of bone marrow mesenchymal stem cells on multiple organ dysfunction syndrome

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