背景:薄型子宫内膜疾病是现阶段临床治疗中的一个难题, 研究发现骨髓间充质干细胞移植技术具有其独特的疗效和优势, 但是关于作用机制方面的研究尚较少开展.目的:基于以HOXA10为中心的调控网络, 探讨骨髓间充质干细胞移植对薄型子宫内膜大鼠的作用机制.方法:将21只SPF级成年雌性SD大鼠 (购于广州中医药大学动物实验中心) 随机分成3组:对照组、模型组、骨髓间充质干细胞组, 每组7只;后2组采用体积分数为95％乙醇充盈宫腔制备薄型子宫内膜模型, 对照组用生理盐水充盈宫腔;抽出乙醇和生理盐水后, 骨髓间充质干细胞组大鼠宫腔内注入第3代骨髓间充质干细胞悬液1m L (细胞浓度为1×1010L-1), 对照组和模型组大鼠宫腔内注入等体积生理盐水.观察2个动情周期后取出子宫, 采用苏木精-伊红染色定量分析内膜厚度, 免疫组化法检测内膜波形蛋白、角蛋白、血管内皮生长因子、白血病抑制因子及整合素αvβ3的表达, q RT-PCR法检测HOXA10、miR-196b基因表达.结果与结论: (1) 模型组和骨髓间充质干细胞组大鼠子宫内膜薄于对照组 (P <0.05), 骨髓间充质干细胞组大鼠子宫内膜厚于模型组 (P <0.05); (2) 模型组和骨髓间充质干细胞组子宫内膜中波形蛋白、角蛋白、血管内皮生长因子、白血病抑制因子及整合素αvβ3蛋白平均吸光度值低于对照组 (P <0.05), 骨髓间充质干细胞组以上5种蛋白平均吸光度值高于模型组 (P<0.05); (3) 模型组和骨髓间充质干细胞组大鼠子宫组织HOXA10基因相对转录水平低于对照组 (P <0.05), mi R-196b基因相对转录水平高于对照组 (P <0.05);骨髓间充质干细胞组HOXA10基因相对转录水平高于模型组 (P<0.05), mi R-196b基因相对转录水平低于模型组 (P<0.05); (4) mi R-196b与HOXA10基因表达呈现负相关 (P <0.05), HOXA10与各蛋白表达呈现不同程度正相关 (P <0.05), miR-196b基因与各蛋白表达呈现不同程度负相关 (P <0.05); (5) 结果提示, 骨髓间充质干细胞移植可一定程度上改善薄型子宫大鼠内膜厚度及相关蛋白水平, 可能原因为mi R-196b负性调节HOXA10基因, HOXA10基因进一步促进血管内皮生长因子、白血病抑制因子及整合素αvβ3蛋白的表达有关.%BACKGROUND: Thin endometrial diseases are a challenge in clinical treatment at present. Scholars have found that bone marrow mesenchymal stem cells (BMSCs) transplantation has its unique curative effect and advantages, but few studies have been conducted on pathway or gene control. OBJECTIVE: To observe the effect of BMSCs transplantation in rats with thin endometrium based on the HOXA10 regulatory network. METHODS: Twenty-one adult female Sprague-Dawley rats of SPF grade (provided by the Animal Experimental Center, Guangzhou University of Chinese Medicine in China) were randomly divided into three groups (n=7/group): control group, model group, and BMSCs group. In the latter two groups, a thin endometrium model was prepared in each rat by filling the uterine cavity with 95％ ethanol. In the control group, normal saline was injected to fill the uterine cavity of rats. After extraction of ethanol or normal saline, the rats in the BMSCs group were injected intrauterinely with 1 mL of BMSCs suspension (1×1010 cells/L), and those in the control and model groups were given the same volume of normal saline. After two estrous cycles, the uterus of each rat was removed. Hematoxylin-eosin staining was used to measure the thickness of the endometrium. Immunohistochemistry was used to detect the expression of vimentin, keratin, vascular endothelial growth factor, leukemia inhibitory factor and integrin αvβ3. qRT-PCR was used to detect the relative transcription of HOXA10 and miR-196 b. RESULTS AND CONCLUSION: (1) Compared with the control group, the endometrial thickness of the rats were significantly thinner in the model and BMSCs groups (P < 0.05), while the endometrial thickness in the BMSCs group was thicker than that in the model group (P < 0.05). (2) The mean absorbance values of endometrial vimentin, keratin, vascular endothelial growth factor, leukemia inhibitory factor and integrin αvβ3 were highest in the control group, higher in the BMSCs group and lowest in the model group, and there were significant differences between groups (P < 0.05). (3) The relative transcript level of HOXA10 gene in the model and BMSCs group was significantly lower than that in the control group, while the relative transcript level of HOXA10 gene in the BMSCs group was significantly higher than that in the model group (P < 0.05). The relative transcript level of miR-196 b in the model and BMSCs groups was significantly higher than that in the control group (P < 0.05), while the relative transcript level of miR-196 b in the BMSCs group was lower than that in the model group (P < 0.05). (4) HOXA10 was negatively correlated with miR-196 b gene, HOXA10 was positively correlated with the protein expression to different extents, and miR-196 b gene was negatively correlated with the protein expression to different extents. These findings suggest that BMSCs transplantation can improve the endometrial thickness and relevant protein levels of thin endometrium rats to some extent, which may be attributed to the negative regulation of HOXA10 gene by miR-196 b, and HOXA10 gene further promotes the expression of vascular endothelial growth factor, leukemia inhibitory factor and integrin αvβ3 proteins.
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