Bone marrow mesenchymal stem cell transplantation combined with Shenghuazhixue decoction in the treatment of postpartum subinvolution of uterus

doi:10.3969/j.issn.2095-4344.2017.21.011

Abstract

Abstract:

BACKGROUND: Bone marrow mesenchymal stem cell (BMSCs) transplantation has been reported to promote the repair of uterine injury; Shenghuazhixue decoction functions to promote blood circulation and remove blood stasis, and dysmenorrhea.

OBJECTIVE: To investigate the effect of BMSCs transplantation combined with Shenghuazhixue decoction on subinvolution of uterus effect and the underlying mechanism.

METHODS: The model of subinvolution of uterus was established in 40 female Sprague-Dawley rats, followed by randomly divided into four groups: control, Shenghuazhixue decoction, BMSCs, and combination groups. The control group received no treatment; at the 1^st day after modeling, 1×10⁷/L BMSCs (1 mL) were injected into the rat myometrium in the BMSCs and combination groups before laparotomy; the rats in the Shenghuazhixue decoction group received no treatment after incision of uterus, and then rinsed using normal saline before closing the abdomen; the Shenghuazhixue decoction and combination groups were given 8 mL/(kg•d) Shenghuazhixue detection via gavage at the 1^st day after modeling, once daily, for consecutive 7 days. Afterwards, the relevant indexes were detected. RESULTS AND CONCLUSION: Compared with the control group, the body weight, uterine wet weight and uterine index in the combined group were significantly decreased after treatment (P < 0.05). The body weight and uterine wet weight showed significant differences between Shenghuazhixue decoction and control groups (P < 0.05). Compared with the control group, the expression levels of intrerleukin-1 and tumor necrosis factor-α were significantly decreased, and the serum level of thromboxane B2 was significantly increased in the Shenghuazhixue decoction group after treatment (P < 0.05). After treatment, the expression levels of intrerleukin-1 and tumor necrosis factor-α were significantly decreased, and the level of vascular endothelial growth factor was significantly increased in the BMSCs group compared with the control group (P < 0.05). Compared with the control group, there was significant decease in the expression levels of intrerleukin-1 and tumor necrosis factor-α, and significant increase in serum level of thromboxane B2 and vascular endothelial growth factor in the combination group (P < 0.01). These results suggest that BMSCs transplantation combined with Shenghuazhixue detection can reduce postpartum body weight, uterine wet weight and index of uterus, and promote uterine involution in female rats, which may be through decreasing the levels of intrerleukin-1 and tumor necrosis factor-α as well as increasing the levels of thromboxane B2 and vascular endothelial growth factor.

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: Uterine Diseases, Bone Marrow, Mesenchymal Stem Cell Transplantation, Tissue Engineering

CLC Number:

R394.2

Chen Ji-long, Feng Jun. Bone marrow mesenchymal stem cell transplantation combined with Shenghuazhixue decoction in the treatment of postpartum subinvolution of uterus[J]. Chinese Journal of Tissue Engineering Research, 2017, 21(21): 3346-3351.

Figures/Tables 3

2.1 实验动物数量分析 40只健康性成熟雌性SD大鼠全部进入结果分析，实验过程无脱失。 2.2 骨髓间充质干细胞的形态在显微镜下观察到原代细胞呈圆形，刚接种时能悬浮在培养瓶中生长(图1A)，接种2 d后细胞呈现梭形分布，沿壁伸展(图1B)；接种3 d后细胞数量增多，围绕中心呈长梭形、放射状排列(图1C)；当细胞融合至80%以上时，细胞呈漩涡状或辐射状平行排列。细胞传代后，贴壁生长速度快，待传至3代以后细胞形态趋于一致。 2.3 骨髓间质干细胞联合生化止血方对大鼠子宫形态学的影响对照组(图2A)内膜不平整，间质炎性细胞增多，腺体增生，肌层变薄，组织水肿，血管扩张，出血明显，平滑肌层亦有散在出血点，间质部分轻度充血水肿，可见中等量散在的含铁血黄素，提示陈旧性出血；生化止血方组(图2B)可见炎细胞浸润，腺体增生，间质、肌层均有少量出血，见少量含铁血黄素，提示陈旧性出血；骨髓间充质干细胞组(图2C)可见较少量炎细胞浸润，腺体增生，间质、肌层均有明显出血，可见少量含铁血黄素，提示陈旧性出血；联合组(图2D)间质部轻度水肿，腺体轻度增生，间质、基层无明显出血，上皮细胞、肌层形态接近正常大鼠子宫组织形态。 2.4 免疫组化法检测各组雌鼠子宫组织白细胞介素1的表达见图3。阳性区域着色呈棕黄色，表达强弱用平均灰度值表示，灰度值越小，表示阳性反应程度越高，说明白细胞介素1表达水平越强。每个标本均连续切片，随机选取4张切片。先在显微镜低倍镜下(×200)对子宫内膜进行定位；再用高倍镜(×400)观察阳性细胞形态和分布，采用MIAS医学图像分析系统进行病理图像分析，计算阳性细胞率、平均灰度值、面密度值。生化止血方组、骨髓间充质干细胞组白细胞介素1表达水平与对照组比较，差异有显著性意义(P < 0.05)；联合组白细胞介素1表达水平与对照组比较，差异有非常显著性意义(P < 0.01)，见表1。 2.5 各组大鼠子宫湿质量、子宫指数的比较与对照组比较，生化止血方组、联合组体质量和子宫湿质量明显减轻，差异有显著性意义(P < 0.05)；生化止血方组、骨髓间充质干细胞组子宫指数与对照组比较，差异无显著性意义(P > 0.05)；联合组子宫指数与对照组比较，差异有显著性意义(P < 0.05)，见表2。 2.6 大鼠血清肿瘤坏死因子α、血栓素B2、血管内皮生长因子水平与对照组比较，生化止血方组血清肿瘤坏死因子α水平明显降低，血栓素B2水平明显升高，差异有显著性意义(P < 0.05)；与对照组比较，骨髓间充质干细胞组血清肿瘤坏死因子α水平明显降低，血管内皮生长因子水平明显升高，差异有显著性意义(P < 0.05)；与对照组比较，联合组血清肿瘤坏死因子α水平明显降低，血栓素B2、血管内皮生长因子水平明显升高，差异有非常显著性意义(P < 0.01)，见表1。"

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