CELL INTELLIGENCE · 肿瘤细胞治疗研究
肿瘤细胞治疗研究
英文原题:Synergistic effects of electroacupuncture and bone marrow mesenchymal stem cell transplantation on rat intrauterine adhesion: an observation.
Synergistic effects of electroacupuncture and bone marrow mesenchymal stem cell transplantation on rat intrauterine adhesion: an observation.
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EA 与 BMSCs 联合应用可协同促进受损子宫内膜修复,改善子宫内膜形态,降低纤维化水平,增强血管再生和基质细胞增殖,提高子宫内膜容受性,最终有利于胚胎着床。
探讨电针联合骨髓间充质干细胞(BMSCs)移植对宫腔粘连(IUA)大鼠子宫内膜的影响,以探究其联合治疗作用的可能机制。
40只成年雌性SD大鼠随机分为对照组、模型组、细胞组和联合组。采用机械搔刮和脂多糖感染双重损伤法建立IUA大鼠模型。造模成功后,于第1、3、7天,模型组大鼠尾静脉注射磷酸盐缓冲液,细胞组大鼠尾静脉注射BMSCs悬液进行BMSCs移植,联合组大鼠接受BMSCs移植联合EA治疗(2 Hz/15 Hz,1-2 mA),取穴“关元”(CV4)、双侧“足三里”(ST36)和“三阴交”(SP6),每日20 min,连续3个动情周期。干预结束后,每组取5只大鼠收集子宫组织。采用苏木精-伊红染色进行组织学分析,评估子宫内膜厚度和腺体数量。采用Masson染色评估子宫内膜纤维化面积。采用免疫组化法检测血管内皮生长因子(VEGF)、增殖细胞核抗原(PCNA)和雌激素受体(ER)的阳性表达。采用Western blot分析检测同源盒A10(HoxA10)和白血病抑制因子(LIF)的蛋白表达,二者均为子宫内膜容受性的关键调节因子。每组剩余5只大鼠与雄性大鼠合笼,通过评估胚胎着床数量评价子宫功能恢复情况。
与对照组比较,模型组子宫内膜变薄(P<0.001),腺体数量减少(P<0.001),子宫内膜纤维化面积增加(P<0.001),VEGF、PCNA、ER阳性表达及HoxA10、LIF表达降低,子宫损伤侧胚胎着床数减少(P<0.001)。与模型组比较,联合组上述指标均出现逆转(P<0.001,P<0.01);细胞组子宫内膜增厚(P<0.001),子宫内膜纤维化面积减少(P<0.001)。与细胞组比较,联合组子宫内膜厚度增加(P<0.01),腺体数量升高(P<0.05),子宫内膜纤维化面积显著减少(P<0.05),子宫内膜中VEGF、PCNA、ER阳性表达及HoxA10、LIF表达增强,子宫损伤侧胚胎着床数显著增加(P<0.001,P<0.05,P<0.01),表明效果优于细胞组。
To investigate the effects of electroacupuncture(EA) combined with bone marrow mesen-chymal stem cells(BMSCs) transplantation on the endometrium of rats with intrauterine adhesions(IUA), so as to explore the possible mechanisms underlying their combined therapeutic effects.
Forty adult female SD rats were randomly divided into control, model, cell, and combined groups. The IUA rat model was established using a dual injury method of mechanical scratching and lipopolysaccharide infection. After successful modeling, on days 1, 3, and 7, rats in the model group received tail vein injection of phosphate buffered solution, while rats in the cell group received tail vein injection of BMSCs suspension for BMSCs transplantation and rats in the combined group received BMSCs transplantation combined with EA treatment (2 Hz/15 Hz, 1-2 mA), targeting the "Guanyuan"(CV4), bilateral "Zusanli"(ST36) and "Sanyinjiao"(SP6) for 20 min daily for 3 consecutive estrous cycles. After intervention, uterine tissue was collected from 5 rats in each group. Histological analysis was performed using hematoxylin and eosin staining to evaluate endometrial thickness and glandular number. Masson staining was used to assess endometrial fibrosis area. Immunohistochemistry was performed to detect the positive expressions of vascular endothelial growth factor(VEGF), proliferating cell nuclear antigen(PCNA), and estrogen receptor(ER). Western blot analysis was conducted to determine the protein expressions of homeobox A10(HoxA10) and leukemia inhibitory factor(LIF), both key regulators of endometrial receptivity. The remaining 5 rats in each group were co-housed with male rats, and the uterine function recovery was evaluated by assessing the number of embryo implantations.
Compared with the control group, the model group showed thinning endometrium( P <0.001), decreased glandular number( P <0.001), increased endometrial fibrosis area( P <0.001), reduced positive expressions of VEGF, PCNA, ER, expressions of HoxA10 and LIF, and decreased embryo implantation number ( P <0.001) on the injured side of the uterus. Compared with the model group, the combined group showed a reversal of the aforementioned indicators( P <0.001, P <0.01) the cell group exhibited thicker endometrium( P <0.001) and reduced endometrial fibrosis area( P <0.001). Compared with the cell group, the combined group showed increased endometrial thickness( P <0.01), elevated glandular number( P <0.05), significantly decreased endometrial fibrosis area( P <0.05), enhanced positive expressions of VEGF, PCNA and ER, expressions of HoxA10 and LIF in the endometrium, and a significant increase in embryo implantation number ( P <0.001, P <0.05, P <0.01) on the injured side of the uterus, indicating better results than the cell group.
The combination of EA and BMSCs synergistically promotes the repair of damaged endometrium, improves endometrial morphology, reduces fibrosis levels, enhances vascular regeneration and matrix cell proliferation, improves endometrial receptivity, which ultimately facilitates embryo implantation. : BMSCs IUA : SD 10 IUA 1 3 7 BMSCs BMSCs BMSCs 20 min 3 5 HE Masson VEGF PCNA ER Western blot A10 HoxA10 LIF 5 : P< 0.001 P< 0.001 P< 0.001 VEGF PCNA ER HoxA10 LIF P< 0.001 P< 0.001 P< 0.01 P< 0.001 P< 0.001 P< 0.01 P< 0.05 P< 0.05 VEGF PCNA ER HoxA10 LIF P< 0.001 P< 0.05 P< 0.01 : BMSCs .
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