坤泰胶囊对卵巢早衰大鼠的治疗作用及其机制研究

Effect and mechanism of Kuntai Capsule on premature ovarian failure in rats

目的观察坤泰胶囊对卵巢早衰模型大鼠的治疗作用,并初步探讨其作用机制。方法将40只SD雌性大鼠随机分为4组:对照组、模型组、坤泰胶囊组、结合雌激素片组,除对照组外,ig 75 mg/kg雷公藤多苷片制备卵巢早衰模型。造模结束后,坤泰胶囊和结合雌激素片组分别以0.6、0.625 g/kg剂量ig给药,连续给药36 d。HE染色观察卵巢形态并进行卵泡计数;采用酶联免疫吸附(Elisa)法检测卵泡刺激素(FSH)、黄体生成素(LH)、血清雌二醇(E2)、及抗缪勒管激素(AMH)水平;实时荧光定量PCR(q RT-PCR)法检测卵巢组织中血管内皮生长因子(VEGF)、碱性成纤维细胞生长因子(b FGF)m RNA的表达变化。结果与对照组比较,模型组卵巢内的始基卵泡、窦前卵泡、窦状卵泡数目明显减少,闭锁卵泡数目明显增多(P<0.05);E2、AMH水平明显降低(P<0.05),FSH、LH水平明显升高(P<0.05);VEGF、b FGF m RNA水平均明显降低(P<0.05)。与模型组比较,坤泰胶囊组卵巢内的始基卵泡、窦状卵泡、窦前卵泡数目明显增多,闭锁卵泡数目明显减少(P<0.05);E2、AMH水平明显升高(P<0.05),FSH、LH水平明显降低(P<0.05);VEGF、b FGF m RNA水平均明显升高(P<0.05)。结论坤泰胶囊通过调节激素水平及上调VEGF、b FGF m RNA表达,修复受损的卵巢组织,促进卵泡的生长发育,从而抑制卵泡过早耗竭。

Objective To explore the effect and mechanism of Kuntai Capsule （KC） on premature ovarian failure in rats. Methods Totally 40 SD female rats were randomly divided into four groups： control group, model group, Kuntai Capsule （KC） group and conjugated estrogens tablets （CET） group. The premature ovarian failure model in rats was made by ig administration of 75 mg/kg tripterygium. After the model was established, rats were continually treated with 0.6 and 0.625 g/kg of KC and CET respectively by ig administration for 36 d. HE staining was used to observe the morphology of ovary and count the number of follicle. The enzyme-linked immunosorbent assay （Elisa） method was used for the detection of follicle stimulating hormone （FSH）, luteinizing hormone （LH）, serum estradiol （E2）, and anti-Mueller tube hormone （AMH） level. The real-time fluorescence quantitative PCR （qRT-PCR） method was used to detect the mRNA expression of vascular endothelial growth factor （VEGF） and basic fibroblast growth factor （bFGF）. Results Compared with control group, the number of primordial follicles, preantral follicles and antral follicles were significantly reduced, the number of atretic follicles was increased （P 〈 0.05）, E2 and AMH significantly decreased （P 〈 0.05）, FSH and LH levels increased significantly （P 〈 0.05）, and the levels of VEGF and bFGF mRNA in model group were significantly decreased （P 〈 0.05）. Compared with model group, the number of primordial follicles, preantral follicles, and antral follicles were significantly increased, the number of atretic follicles was significantly reduced （P 〈 0.05）, E2 and AMH significantly increased （P 〈 0.05）, FSH and LH levels significantly decreased （P 〈 0.05）, and the levels of VEGF and bFGF mRNA in KC group were significantly increased （P 〈 0.05）. Conclusion By regulating the level of hormone and up-regulating the expression of VEGF and bFGF, KC can repair the damaged ovarian tissue and promote the growth and development of the follicle, so as to inhibit the premature exhaustion of mRNA.