摘要
目的：采用雌-孕激素负荷法建立大鼠子宫肌瘤模型,研究妇瘤方对子宫肌瘤大鼠子宫形态学、血清E2、P浓度及肌瘤组织中TGF-β1、BFGF表达水平的影响,探讨其对子宫肌瘤的治疗作用。
方法：取未孕SD系清洁级雌性大鼠50只,体重202.10±8.32g,以雌-孕激素负荷法建立子宫肌瘤模型,另取10只肌注生理盐水做空白对照。造模结束后将50只造模大鼠随机分为5组,分别为模型对照组、宫瘤宁胶囊组、妇瘤方低、中、高剂量组。造模结束第二天,各治疗组开始灌胃给药,空白组、模型组灌给蒸馏水,连续28天后处死大鼠,观察子宫形态学变化,放射免疫法测定大鼠血清雌二醇(E2)、孕酮(P)浓度,免疫组化法观察大鼠子宫平滑肌细胞中转化生长因子-β1(TGF-β1)、碱性成纤维细胞生长因子(BFGF)的表达。
结果：
1.模型组大鼠子宫各项指标明显高于空白组(P<0.01),给予药物干预后,各治疗组子宫重量、子宫系数及子宫平滑肌厚度均减小,与模型组比较有显著性差异(P<0.01)。
2.子宫平滑肌层病理组织学改变为出现多发性、局灶性的病变,肌层增厚,肌细胞增大,排列紊乱,有双核并偶见核分裂相出现,且伴有结缔组织增多,间质细胞浓染而稍瘦小,表明局部平滑肌增殖活跃,和临床子宫肌瘤病理组织学很相似。
3.模型组大鼠血清E2、P水平明显高于正常大鼠(P<0.01)。治疗各组大鼠血清E2、P浓度均有不同程度下降,与模型组比较有统计学意义(P<0.05或P<0.01)。
4.模型组大鼠TGF-β1表达水平明显高于空白对照组(P<0.01)；经药物干预,各治疗组TGF-β1表达均有所降低(P<0.05或P<0.01)；各治疗组间比较差别无统计学意义(P>0.05)。
5.模型组大鼠BFGF表达水平较空白对照组明显增高(P<0.01)；经药物干预,各治疗组表达水平均有所降低(P<0.05或P<0.01)；各治疗组间比较差别无统计学意义(P>0.05)。
结论：
1.采用雌、孕激素负荷法成功建立了大鼠子宫肌瘤模型。
2.妇瘤方可明显降低雌、孕激素负荷大鼠子宫重量、子宫系数及子宫平滑肌厚度,具有抑制子宫平滑肌瘤样增生的作用。
3.妇瘤方可明显降低雌、孕激素负荷大鼠血清E2.P水平。
4.TGF-β1、BFGF在子宫肌瘤的发生发展中起重要作用,参与子宫肌瘤的发病机制。
5.妇瘤方能够抑制子宫平滑肌细胞中TGF-β1、BFGF表达,这可能是妇瘤方抑制子宫肌瘤发展的重要机制之一。
Object:To study the effect of FuLiu Formula on the Morphology of Uterus, the serum concentration of estradiol and progesterone,the expression of Transforming growth factor-β1(TGF-β1) and Basic fibroblast growth factor(BFGF) in Uterine Leiomyomas Model Rats with estrogen and progesterone. Explore the treatment acts of FuLiu Formula on uterine leiomyomas.
Methods:50 SD female、non-pregnant rats were choosed as researeh object. All of them were established the animal models of the Uterine Leiomyomas by Loading the Rats with Estrogen and Progesterone.And the other 10 rats were loaded with normal saline as control contrasting group. After that, all model rats were randomly divided into 5 groups:modle group, Gong Liu Ning capsule group, lower dosage group of Fu Liu Formula, medium dosage group of Fu Liu Formula, higher dosage group of Fu Liu Formula. The second day,every treatment group were fed with drug, control contrasting group and modle group were fed with distilled water. After 28 days, they were killded.The morphology changes of Uterus were observed.The serum concentrations of estradiol and progesterone were measured by radioim-noassay(RIA), Expressions of transforming growth factor-β1 and basic fibroblast with factor in smooth muscle cells were detected by mmunohistochemistry.
Results:
1.Compared with the control group, the weight of uterines, uterus index, thickness of womb smooth muscle in modle group was changed significantly(P<0.01).Compared with the model group, weight of uterines, uterus index, thickness of womb smooth muscle of every treatment group were dropped obviously(P<0.01).
2.The myometrium of treated rats showed the proliferative features, both multiple and focal.The myometrium were thickened and the smooth muscle cells enlarged. Binuclei and the nucleiin mitotic phase could be easily observed in the cells.And more connective materials constructed the extracellular matrix.All these mentioned tendency could be characterized by excessively focal proliferation,which was similar to those of patients with uterine leiomyoma.
3.Compared with the control group, the concentration of estradiol and progesterone in modle group was increased obviously(P<0.01). Compared with the model group, the concentration of estradiol and progesterone in every treatment group was dropped significantly (P<0.05 or P<0.01).
4.Compared with the control group, the expression of TGF-β1 positive cells were increased obviously(P<0.01).And in every treatment group they was dropped significantly(P<0.05 or P<0.01). But compared with each other, it had none statistical significance(P>0.05).
5.Compared with the control group, the expression of BFGF positive cells were increased obviously(P<0.01).And in every treatment group they was dropped significantly(P<0.05 or P<0.01). But compared with each other, it had none statistical significance(P>0.05).
Conclution:
1.Animal models of the Uterine Leiomyomas were established successfully by loading the rats with Estrogen and Progesterone.
2.FuLiu Formula can reduce the weight of uterines, uterus index, thickness of womb smooth muscle of rats'Uterine with Estrogen and Progesterone obviously.
3.FuLiu Formula can reduce the concentration of estradiol and progesterone of rats'Uterine with Estrogen and Progesterone obviously.
4.The concentration of Transforming growth factor-β1 and Basic fibroblast growth factor play an important part in the growth of Uterine Leiomyomas, FuLiu Formula.
5. FuLiu Formula can reduce the expression of Transforming growth factor-β1 and Basic fibroblast growth factor,which was demonstrated may be one of reasons to treat Uterine Leiomyomas.
引文
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