Leonurine combined with bone marrow mesenchymal stem cell transplantation in the treatment of intrauterine adhesions

doi:10.3969/j.issn.2095-4344.2016.32.009

Abstract

Abstract:

BACKGROUND: Increasing evidence has demonstrated that bone marrow mesenchymal stem cell (BMSC) transplantation can promote skin, liver and lung repair in animal models; and Leonurus sibiricus L. has the ability of promoting blood circulation and regulating menstruation.
OBJECTIVE: To investigate the therapeutic effect of BMSC transplantation with leonurine on intrauterine adhesions (IUA) in rabbits and the relevant mechanism of action.
METHODS: After modeled using dual injury method, rabbit models of IUA were randomly divided into five groups: sham-operated group (sham), model group (IUA), BMSC transplantation group, leonurine treatment group and combined treatment group (BMSCs+leonurine). Rabbits in the sham group were only given normal saline rinsing after hysterotomy, while those in the latter three groups were correspondingly given intrauterine BMSC transplantation or/and intragastric administration of 4 mg/kg leonurine for 14 days. Morphological changes of the endometrium were observed using hematoxylin-eosin staining, and expression levels of transforming growth factor β protein, Smad3 protein and interferon-γ mRNA were detected using immunohistochemical staining and real-time fluorescence quantitative PCR, respectively.
RESULTS AND CONCLUSION: Compared with the model group, the degree of IUA was all significantly improved in the other groups, especially in the combined treatment group. Moreover, BMSC transplantation, leonurine treatment and their combined use all could inhibit IUA-induced increase of transforming growth factor β and Smad3 protein expression and IUA-induced decrease of interferon-γ mRNA level. Importantly, all these alternations were much more pronounced in the combined treatment group. Our results show that the combined use of BMSC transplantation and leonurine treatment can exert a synergistic effect in the improvement of IUA through the transforming growth factor β/Smad3 pathway.

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: Mesenchymal Stem Cells, Leonurus, Uterus, Transforming Growth Factor beta, Smad3 Protein, Tissue Engineering

CLC Number:

R394.2

Wang Bing-yu, Ji Xia. Leonurine combined with bone marrow mesenchymal stem cell transplantation in the treatment of intrauterine adhesions[J]. Chinese Journal of Tissue Engineering Research, 2016, 20(32): 4771-4777.

Figures/Tables 2

2.1 实验动物数量分析实验选用西兰大白兔40只，分为5组，实验过程无脱失，全部进入结果分析。 2.2 骨髓间充质干细胞形态显微镜下观察到原代细胞呈圆形，刚接种时悬浮在培养瓶中，24 h后细胞呈梭形，沿壁伸展；3 d后细胞数量增多，呈长梭形并围绕中心呈放射状排列；当细胞融合至80%以上时，细胞平行排列成漩涡状或辐射状。细胞传代后，生长速度加快，迅速贴壁，待传至3代以后细胞形态趋于一致。见图1。 2.3 子宫内膜形态学变化假手术组子宫腔表面覆盖柱状上皮，内膜层腺体丰富；宫腔粘连组纵向剖开宫腔，可见膜样粘连带形成，显微镜下观察宫腔出现纤维性粘连带，未粘连区几乎由无功能的扁平上皮细胞覆盖，上皮层下腺体极少；骨髓间充质干细胞组宫腔部分闭锁，表面可见纤维性粘连带，未粘连区有扁平上皮细胞覆盖，上皮层下腺体极少；益母草碱组可见大量膜样粘连带，粘连区大部分覆盖扁平上皮细胞；联合组大部分宫腔表面上皮再生，上皮细胞呈立方形，再生上皮层下见少量腺体。见图2。 2.4 各组组织中转化生长因子β、Smad3的蛋白表达变化宫腔粘连组组织中转化生长因子β、Smad3的蛋白表达呈强阳性；与宫腔粘连组相比，益母草碱组、骨髓间充质干细胞组与联合组组织中转化生长因子β、Smad3的蛋白表达水平显著降低(P < 0.05)，其蛋白表达的强弱程度为：益母草碱组>骨髓间充质干细胞组>联合组。见图3，4。 2.5 各组组织中干扰素γ的mRNA表达变化如图5所示，宫腔粘连组组中干扰素γ的mRNA表达呈阴性；与宫腔粘连组相比，益母草碱组、骨髓间充质干细胞组与联合组组织中干扰素γ的mRNA表达水平显著升高(P < 0.05)，其蛋白表达的强弱程度为：益母草碱组<骨髓间充质干细胞组<联合组。"

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