摘要
目的探讨橘络饮联合乳核散对乳腺增生模型大鼠乳腺增生的影响及其机制。方法SPF级7~8周龄SD雌性大鼠40只,随机分为对照组(A组)、乳腺增生组(B组)、乳癖消治疗组(C组)、橘络饮联合乳核散治疗组(D组),每组各10只。B、C、D组肌肉注射苯甲酸雌二醇和孕酮构建乳腺增生模型,A组注射等剂量生理盐水。造模成功后,D组灌胃橘络饮汤剂并于第2、3对乳房处用弹力带包裹乳核散浸膏30 d;C组灌胃乳癖消溶液,A、B组灌胃等体积生理盐水,而后三组均用弹力带包裹第2、3对乳房30 d。药物处理结束后测量各组大鼠第2对乳房的乳头高度和乳房直径;通过酶联免疫吸附实验(ELISA)检测各组大鼠血清中雌二醇(E2)、孕酮、卵泡素(FSH)和促黄体激素(LH)的水平;通过苏木精-伊红(HE)染色和Masson染色观察各组大鼠乳腺组织的病理变化;通过蛋白免疫印迹实验检测各组大鼠乳腺组织中磷脂酰肌醇3激酶(PI3K)/丝氨酸-苏氨酸激酶(AKT)/B细胞淋巴瘤因子-2(Bcl-2)信号通路和细胞外调节蛋白激酶(ERK)信号通路相关蛋白的相对表达水平;通过HE染色观察各组大鼠肝脏和肾脏组织形态。结果与B组相比,C、D组大鼠的乳头高度和乳房直径均显著降低,其中D组的乳头高度显著低于C组(F=104.10、172.70,t=2.54~12.24,P<0.05)。ELISA检测结果显示,与B组相比,C和D组大鼠血清中E2和FSH的水平显著降低,孕酮和LH的水平显著升高,其中D组大鼠血清中孕酮和LH的水平显著高于C组(F=16.46~149.10,t=4.53~15.25,P<0.05)。HE和Masson染色结果显示,与B组相比,C、D组大鼠乳腺组织内腺体均显著减少,导管扩张及导管上皮细胞增生均显著缓解,导管内分泌物和胶原沉积均显著减少,其中D组上述病理特征的改善较C组更加显著。蛋白免疫印迹实验结果显示,与B组相比,C、D组大鼠乳腺组织中p-AKT/AKT、Bcl-2/Bax、p-ERK/ERK的比值和ERα的相对表达水平均显著降低,其中D组大鼠乳腺组织中ERα的相对表达水平较C组更低(F=21.37~205.50,t=4.94~30.51,P<0.05)。HE染色结果显示,各组大鼠的肝脏和肾脏组织形态均无明显异常。结论橘络饮联合乳核散能够缓解乳腺增生模型大鼠的乳房增大、体内性激素紊乱和乳腺组织的病理改变,其机制可能与抑制PI3K/AKT/Bcl-2和ERK信号通路的活化有关。
Objective To investigate the effect and mechanism of Juluo Decoction combined with Ruhe Powder in the treatment of rat model of hyperplasia of mammary glands.Methods A total of 40 specific pathogen-free female Sprague-Dawley rats,aged 7-8 weeks,were randomly divided into control group(group A),hyperplasia of mammary glands group(group B),Rupixiao treatment group(group C),and Juluo Decoction+Ruhe Powder treatment group(group D),with 10 rats in each group.The rats in groups B,C,and D were given intramuscular injection of estradiol benzoate and progesterone to establish a model of hyperplasia of mammary glands,and those in group A were given an equal volume of saline.After successful modeling,the rats in group D were given Juluo Decoction by gavage,with the second and third pairs of mammary glands wrapped with elastic bands containing Ruhe Powder extract for 30 days;the rats in group C were given Rupixiao solution by gavage,while those in groups A and B were given an equal volume of normal saline by gavage,and for the rats in groups A,B,and C,the second and third pairs of mammary glands were wrapped with elastic bands for 30 days.After the end of drug treatment,nipple height and breast diameter were measured for the second pair of mammary glands in each group;ELISA was used to measure the serum levels of estradiol(E2),progesterone,follicle-stimulating hormone(FSH),and luteinizing hormone(LH);HE staining and Masson staining were used to observe the pathological changes of mammary gland tissue in each group;Western blotting was used to measure the relative expression levels of proteins associated with the phosphatidylinositol 3-kinase(PI3K)/serine-threonine kinase(AKT)/B-cell lymphoma-2(Bcl-2)signaling pathway and the extracellular signal-regulated kinase(ERK)signaling pathway in mammary gland tissue;HE staining was used to observe the histomorphology of the liver and the kidney.Results Compared with group B,groups C and D had significant reductions in nipple height and breast diameter,and group D had a significantly lower nipple height than group C(F=104.10,172.70,t=2.54-12.24,P<0.05).ELISA showed that compared with group B,groups C and D had significant reductions in the serum levels of E2 and FSH and significant increases in the serum levels of progesterone and LH,and group D had significantly higher serum levels of progesterone and LH than group C(F=16.46-149.10,t=4.53-15.25,P<0.05).HE staining and Masson staining showed that compared with group B,groups C and D had significant reductions in the glands in mammary gland tissue,significant alleviation of duct dilation and duct epithelial cell proliferation,and significant reductions in duct secretions and collagen deposition,and compared with group C,group D had significantly greater improvements in these pathological features.Western blotting showed that compared with group B,groups C and D had significant reductions in the ratios of p-AKT/AKT,Bcl-2/Bax,and p-ERK/ERK and the relative expression level of ERαin mammary gland tissue,and compared with group C,group D had a significantly lower relative expression level of ERαin mammary gland tissue(F=21.37-205.50,t=4.94-30.51,P<0.05).HE staining showed that there were no obvious abnormalities in the histomorphology of the liver and the kidney.Conclusion Juluo Decoction combined with Ruhe Powder can alleviate breast enlargement,sex hormone disorder,and the pathological changes of breast tissue in a rat model of hyperplasia of mammary glands,possibly by inhibiting the activation of the PI3K/AKT/Bcl-2 and ERK signaling pathways.
作者
聂卫红
刘鸿禧
孙国欣
刘诚聪
邹晓
NIE Weihong;LIU Hongxi;SUN Guoxin;LIU Chengcong;ZOU Xiao(Qingdao Medical College of Qingdao University,Qingdao 266071,China)
出处
《精准医学杂志》
2025年第2期131-136,共6页
Journal of Precision Medicine
基金
青岛市市南区科技基金项目(2022-2-023-YY)。
