摘要
目的探讨补肾养阴法协同骨髓间充质干细胞(BMSCs)移植治疗卵巢早衰(POF)的可行性。方法以连续腹腔注射环磷酰胺(CTX)14 d的方法建立卵巢早衰模型。其后分别用BMSCs移植及BMSCs移植联合补肾养阴中药灌胃法治疗卵巢早衰,以青年大鼠为空白对照,治疗28 d后,Elisa法检测外周血雌二醇(E2)、卵泡刺激素(FSH);Tunel法检测卵巢颗粒细胞凋亡;Western-blot法检测Bcl-2相关X蛋白(Bax)mRNA、B淋巴细胞瘤-2基因mRNA(Bcl-2)相对表达量。结果 A组(空白对照组)注射生理盐水前后激素水平无明显变化;建模后14 d,与A组比较,B组(模型组)大鼠血E2水平显著降低(P<0.01),而FSH水平明显升高(P<0.01);与B组比较,治疗后C组(干细胞干预组)及D组(干细胞+补肾养阴药物干预组)E2水平呈逐渐上升趋势(P<0.01),FSH水平逐渐下降(P<0.01),其中D组变化幅度明显高于C组(P<0.05);Tunel法检测细胞凋亡结果显示,A组染色较少,B组最多,C、D 2组染色程度介于A、B组之间,C、D 2组之间染色程度差异不明显。Western-blot检测结果显示,Bax mRNA在B组表达明显增多(P<0.01),用药后,C、D 2组Bax mRNA有所下降,尤其以D组明显,与B组比较,差异有显著性意义(P<0.05)。Bcl-2 mRNA在B组表达最少(P<0.01);用药后,C、D 2组Bcl-2 mRNA表达均有所增加,与B组比较,差异有极显著性意义(P<0.01)。结论 BMSCs移植联合补肾养阴中药灌胃法可改善化疗后(POF)大鼠卵巢功能,其疗效优于单纯使用BMSCs移植。
Objective To explore the feasibility of Zuogui pill（ ZGP）,a traditional Chinese medicine formulated by TCM therapy of tonifying kidney-yin,with the therapy of bone marrow mesenchymal stem cells（ BMSC） transplantation in premature ovarian failure（ POF） induced by chemotherapy.Methods The rat model of POF was established by intraperitoneal injection of cyclophosphamide（ cytoxan,CTX） for consecutive 14 d. All the PDF rats were divided into model group,BMSC transplantation（ BMSCT） group and BMSCT combined with ZGP（ BMSCT ＋ ZGP） group,and the healthy young rats were included in control group. The levels of estradiol（ E2）,follicle-stimulating hormone（ FSH） in peripheral blood were determined by using ELISA,the ovarian granulosa cell apoptosis were tested by Tunel assay,and the mRNA expressions of Bcl-2 Associated X（ Bax） and Bcell lymphoma-2（ Bcl2） were examined by Western blot. Results After successful modeling,the level of E2 in peripheral blood in the model group was much lower than that in the control group,and the level of FSH in peripheral blood was much higher（ both P 0. 01）. Compared with model group,the level of E2 increased gradually while FSH decreased guadually in both BMSCT group and BMSCT ＋ ZGP group（ P 0. 01）,and the changes of E2 and FSH were more obvious in BMSCT ＋ ZGP group than those in BMSCT group. Tunel showed that the number of cells decreased in both BMSCT group and BMSCT ＋ ZGP group,compared with model group,but there were no significant difference between the two treatment groups. The mRNA expressions of Bax were lower in both treatment groups than that of model group,and the significant decreasing was found in BMSCT ＋ ZGP group compared with model group（ P 0. 05）. The mRNA expressions of Bcl2 increased significantly after BMSCT and ZGP treatment compared with model group（ P 0. 01）. Conclusion BMSCT combined with ZGP could improve ovarian function in POF rats induced by chemotherapy,and the effect was better than that of BMSCT on
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2015年第11期735-739,I0001,共6页
Journal of Beijing University of Traditional Chinese Medicine
基金
国家自然科学基金青年基金资助项目(No.30901917)
国家自然科学基金面上资助项目(No.81373805)
关键词
卵巢早衰
骨髓间充质干细胞
补肾养阴法
大鼠
premature ovarian failure
bone marrow mesenchymal stem cell
TCM therapy of tonifying kidney-yin
rats
