麦门冬汤对大鼠骨髓间充质干细胞向肺泡上皮细胞分化的影响
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摘要
[目的]观察麦门冬汤含药血清对促进大鼠骨髓间充质干细胞(BMSC)向肺泡上皮细胞分化的影响。[方法]分别予麦门冬汤大鼠血清、肺纤维化模型大鼠血清、肺纤维化模型大鼠+麦门冬汤血清对绿色荧光蛋白标记的骨髓间充质干细胞(GFP-BMSC)进行干预14 d,通过免疫组化、实时荧光定量聚核酶链反应(qPCR)方法,检测各组GFPBMSC细胞肺表面活性蛋白(SP-C)、水通道蛋白-5(AQP-5)蛋白及mRNA的表达。[结果]模型组、模型+麦门冬汤组SP-C、AQP5蛋白表达明显高于DMEM对照组、麦门冬汤组(P<0.01),其中模型+麦门冬汤组SP-C、AQP5表达明显高于模型组(P<0.01)。[结论]麦门冬汤在肺纤维化环境中可促进GFP-BMSC向肺泡上皮细胞分化,该作用可能是其改善肺纤维化的重要机制。
[Objective] To observe the effect of Maimendong Decoction on bone marrow mesenchymal stem cells(BMSC) to differentiate into alveolar epithelial cells of rats. [Methods] Maimendong Decoction rat serum, rat model of pulmonary fibrosis in rats serum and rat model of pulmonary fibrosis+Maimendong Decoction serum were given on GFP-BMSC. After 14 days, to detect the expression of SP-C,AQP-5 protein and mRNA in GFP-BMSC cells by immunohistochemistry and qPCR method. [Results] The expression of SP-C and AQP-5 proteins in the model group, model+Maimendong Decoction group was significantly higher than that of DMEM control group, Maimendong Decoction group(P<0.01); while the model+Maimendong Decoction group SP-C, AQP-5 proteins expression was significantly higher than the model group(P<0.01). [Conclusion] Maimendong Decoction can promote GFP-BMSC cells into alveolar epithelial cells in pulmonary fibrosis, which may be one of the important mechanisms to improve pulmonary fibrosis.
[Objective] To observe the effect of Maimendong Decoction on bone marrow mesenchymal stem cells(BMSC) to differentiate into alveolar epithelial cells of rats. [Methods] Maimendong Decoction rat serum, rat model of pulmonary fibrosis in rats serum and rat model of pulmonary fibrosis+Maimendong Decoction serum were given on GFP-BMSC. After 14 days, to detect the expression of SP-C,AQP-5 protein and mRNA in GFP-BMSC cells by immunohistochemistry and qPCR method. [Results] The expression of SP-C and AQP-5 proteins in the model group, model+Maimendong Decoction group was significantly higher than that of DMEM control group, Maimendong Decoction group(P<0.01); while the model+Maimendong Decoction group SP-C, AQP-5 proteins expression was significantly higher than the model group(P<0.01). [Conclusion] Maimendong Decoction can promote GFP-BMSC cells into alveolar epithelial cells in pulmonary fibrosis, which may be one of the important mechanisms to improve pulmonary fibrosis.
引文
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[26]宋殿荣,王雅楠,张崴,等.小半夏加茯苓汤含药血清对小鼠胚胎干细胞体外增殖及分化的影响[J].天津中医药,2011,28(6):486-488.
[27]张文信,刘金文,曹学伟,等.金匮肾气丸含药血清对体外兔BMSCs增殖细胞周期及凋亡的影响[J].辽宁中医杂志,2011,38(8):1511-1514.
[28]詹菲,徐志伟,黄进,等.左归丸对衰老大鼠骨髓间充质干细胞增殖的影响[J].中华中医药杂志,2015,30(7):2518-2521.
[2]Mora AL,Rojas M.Aging and lung injury repair:a role for bone marrow derived mesenchymal stem cells[J].Journal of cellular biochemistry,2008,105(3):641-647.
[3]赵峰,李圣青,遆新宇,等.骨髓间充质干细胞对肺损伤大鼠转化生长因子β及单核细胞趋化蛋白1的影响[J].中国组织工程研究与临床康复,2008,12(29):5627-5630
[4]Liu Q,Jiang JX,Liu YN,et al.Grape seed extract ameliorates bleomycin-induced mouse pulmonary fibrosis[J].Toxicology Letters,2017(273):1-9.
[5]黄云鉴,龚婕宁.中医治疗肺纤维化方药规律的文献分析[J].中国实验方剂学杂志,2016,22(15):206-210.
[6]赵庆波,吴洪军,迟铁汉,等.特发性肺纤维化中西医发病机制研究进展[J].河北医学,2016,22(11):1910-1912.
[7]马锦地.基于现代名老中医经验的肺痿辨证与方药规律研究[D].郑州:河南中医学院,2015.
[8]瓮恒,曲中平.麦门冬汤加减治疗特发性肺间质纤维32例临床观察[J].国医论坛,2008,23(1):6-7
[9]王洪,李霞,辛大永,等.养阴益气合剂对特发性肺间质纤维化气阴两虚证患者生活质量的影响[J].中医杂志,2016,57(9):767-770.
[10]宋建平,刘方州,李瑞琴,等.栝蒌薤白汤、麦门冬汤及肾气丸对平阳霉素所致肺纤维化的影响[J].国医论坛,2001,16(4)40-41.
[11]李瑞琴,宋建平,张瑞,等.麦门冬汤对肺纤维化模型早期阶段肺组织中TNF-α、IL-10表达的影响[J].中医研究,2009,22(9):10-13.
[12]宋建平,李伟,李瑞琴,等.《金匮要略》不同方药对肺纤维化大鼠模型早期阶段(1~14 d)的影响[J].中国中医基础医学杂志,2009,(15)6:432-434.
[13]杨美凤,谢忠礼,宋建平.麦门冬汤对大鼠肺纤维化形成阶段肺病理损害及下丘脑多巴胺、去甲肾上腺素、五羟色胺的干预作用[J].中国医药导报,2012,9(12):36-38.
[14]杜光明.麦门冬汤对肺纤维化大鼠肺肠组织形态学、TNF-α等的影响[D].郑州:河南中医学院,2015.
[15]赵晨宇,祝勇,周济谱,等.养阴益气合剂对博来霉素所致肺纤维化大鼠的影响[J].中国中医药信息杂志,2015,22(9):63-67.
[16]王力,吴鑫,卢新政,等.人参皂苷Rg1对培养大鼠骨髓间充质干细胞增殖影响的机制研究[J].中国药理学通报,2007,23(11):1840-1844.
[17]李佳玮,张密霞,周涛,等.人参皂苷Rg1干预骨髓间充质干细胞转化为多潜能干细胞关键基因m RNA的表达[J].中国组织工程研究与临床康复,2011,15(32):6032-6035.
[18]邬伟,杨景全,刘宝华,等.人参皂苷Rg1对痴呆大鼠脑匀浆中BMSCs的神经性分化作用[J].中风与神经疾病杂志,2011,28(3):204-207.
[19]郑国庆,王小同,邬伟,等.人参总皂苷体外诱导大鼠骨髓间充质干细胞分化为神经元样细胞[J].中华中医药学刊,2008,26(6):1257-1259.
[20]Shea KH,Tsui YP,Ying SC,et al.Bone marrow-derived Schwann cells achieve fate commitment-a prerequisite for remyelination therapy.[J].Experimental Neurology,2010,224(2):448-458.
[21]Kim JY,Oh CH,Huang X,et al.Improvement in sensory function via granulocyte-macrophage colony-stimulating factor in rat spinal cord injury models[J].Journal of Neurosurgery Spine,2013,18(1):69-75.
[22]Chen X,Li Y,Wang L,et al.Ischemic rat brain extracts induce human marrow stromal cell growth factor production.[J].Neuropathology,2002,22(4):275-279.
[23]Ji WH,Choi D,Min YL,et al.Bone Marrow-Derived Mesenchymal Stem Cells Improve Diabetic Neuropathy by Direct Modulation of Both Angiogenesis and Myelination in Peripheral Nerves[J].Cell Transplantation,2016,25(2):313.
[24]Mezey E.The therapeutic potential of bone marrow-derived stromal cells[J].Journal of cellular biochemistry,2011,112(10):2683-2687.
[25]方健康,周轶平,李玛琳.中药对骨髓间充质干细胞增殖、凋亡及分化的影响研究进展[J].中国中药杂志,2014,39(15):2834-2837.
[26]宋殿荣,王雅楠,张崴,等.小半夏加茯苓汤含药血清对小鼠胚胎干细胞体外增殖及分化的影响[J].天津中医药,2011,28(6):486-488.
[27]张文信,刘金文,曹学伟,等.金匮肾气丸含药血清对体外兔BMSCs增殖细胞周期及凋亡的影响[J].辽宁中医杂志,2011,38(8):1511-1514.
[28]詹菲,徐志伟,黄进,等.左归丸对衰老大鼠骨髓间充质干细胞增殖的影响[J].中华中医药杂志,2015,30(7):2518-2521.