染料木黄酮对小鼠子宫内膜异位症抑制作用的研究

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目的:探讨染料木黄酮对小鼠子宫内膜异位症(endometriosis,EMs)的抑制作用及其机制。方法:选择70只ICR雌性小鼠分为对照组和内膜异位组,10只小鼠只经过假手术作为对照组(A组),60只小鼠采用自体腹壁下移植的方法,制备子宫内膜异位症的动物模型,同时皮下注射17β-雌二醇0.5μg/kg,每3天1次共3周。3周后处死20只小鼠,检测移植物体积和病理学检查。剩余的小鼠随机分为B组(模型组,n=10),染料木黄酮组分为C组(低剂量组,0.35 mg/kg,n=10)、D组(中剂量组,3.5 mg/kg,n=10)和E组(高剂量组,35 mg/kg,n=10)。染料木黄酮组用3个不同剂量1次/d,皮下注射,共12周,A和B组用芝麻油每次0.05 ml,每天1次,共12周。第12周末,全部实验动物眼球采血后处死,检测异位病灶体积后做病理检查,采用ELISA法检测血清中抗子宫内膜抗体(antiendometrial antibody,EMAb)、白细胞介素-6(inter-leukin-6,IL-6)和肿瘤坏死因子-α(tumour necrosis factor-alpha,TNF-α)含量。结果:本实验小鼠子宫内膜异位模型的形成率为68.3%(41/60)。C组移植物体积治疗前、后比较差异无统计学意义(P>0.05)。D组〔(135.6±15.1)mm3、(61.4±9.0)mm3〕、E组〔(137.8±15.6)mm3、(11.6±3.3)mm3〕治疗前、后移植物体积比较明显减小(P<0.05),E组与D组比较移植物体积差异有统计学意义(P<0.01)。B组与A组比较血清EMAb、IL-6和TNF-α的含量明显升高(P<0.05)。C组与B组比较血清中IL-6、TNF-α的含量差异无统计学意义(P>0.05),D组〔(62.54±5.10)pg/ml、(29.89±3.54)pg/ml〕和E组〔(31.18±2.97)pg/ml、(19.34±2.85)pg/ml〕与B组〔(125.04±3.23)pg/ml、(49.00±5.15)pg/ml〕比较血清中IL-6、TNF-α的含量明显降低(P<0.05),D组与E组比较差异有统计学意义(P<0.01)。结论:采用自体腹壁下移植同时皮下注射雌二醇的方法制备EMs动物模型方法简单,是研究EMs较理想的动物模型。染料木黄酮对小鼠子宫内膜异位移植体有明显的退化作用,其作用可能是与雌二醇竞争性结合ER、降低雌激素水平以及细胞因子的作用有关。 Objective: To investigate the inhibitory effect of genistein on mouse endometriosis (EMs) and its mechanism. Methods: Seventy ICR female mice were divided into control group and endometriosis group, 10 mice were sham operated as control group (group A), 60 mice were transplanted into the uterus Animal models of endometriosis, while subcutaneous injection of 17β-estradiol 0.5μg / kg, once every 3 days for 3 weeks. After 3 weeks, 20 mice were sacrificed and the graft volume and pathological examination were examined. The remaining mice were randomly divided into group B (model group, n = 10), genistein component C (low dose group, 0.35 mg / kg, n = 10), group D / kg, n = 10) and group E (high dose, 35 mg / kg, n = 10). The genistein group was treated with 3 different doses of 1 / d subcutaneously for 12 weeks. Groups A and B were treated with sesame oil 0.05 ml once daily for 12 weeks. At the end of the twelfth week, all the animals were sacrificed after eyeball blood sampling. The volume of ectopic lesions was detected and pathological examination was performed. The serum levels of anti-endometrial antibody (EMAb), inter-leukin- 6, IL-6) and tumor necrosis factor-alpha (TNF-α). Results: The formation rate of mouse endometriosis model in this experiment was 68.3% (41/60). The volume of graft in group C had no significant difference before and after treatment (P> 0.05). The volume of graft in group D was (135.6 ± 15.1) mm3, (61.4 ± 9.0) mm3 in group D compared with that in group E [(137.8 ± 15.6) mm3, (11.6 ± 3.3) mm3] ), There was a significant difference in the volume of grafts between group E and group D (P <0.01). The levels of serum EMAb, IL-6 and TNF-α in group B were significantly higher than those in group A (P <0.05). The serum levels of IL-6 and TNF-α in group C and group B had no significant difference (P> 0.05), but in group D (62.54 ± 5.10) pg / ml and (29.89 ± 3.54) pg / Serum levels of IL-6, IL-6 and IL-6 in group E were significantly higher than those in group B [(31.18 ± 2.97) pg / ml and (19.34 ± 2.85) pg / (P <0.05). There was significant difference between group D and group E (P <0.01). Conclusion: It is a simple method to prepare animal model of EMs by subcutaneous abdominal subcutaneous transplantation with subcutaneous injection of estradiol. It is a better animal model to study EMs. Genistein has a significant degenerative effect on mouse endometriosis, and its role may be related to the competitive binding of estradiol ER, reduce the level of estrogen and the role of cytokines.
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