摘要
目的:研究乳癖散结汤对乳腺增生(MGH)大鼠的治疗作用及可能的作用机制。方法:MGH大鼠模型制作成功后,随机分为正常对照组、模型对照组、乳癖散结汤34、68 g/kg组、他莫昔芬0.004 g/kg组;大鼠分别灌胃给予相应的药物。给药8 w后,进行大鼠行为学观察、乳房直径和乳头高度测量,测定血浆中T淋巴细胞亚群(CD^(3+)、CD^(4+)、CD^(8+))、血清性激素[雌二醇(E2)、孕酮(P)、睾酮(T)、黄体生成激素(LH)、卵泡刺激素(FSH)、泌乳激素(PRL)]含量,取乳腺进行乳腺组织细胞线粒体细胞色素C(Cyto C)释放、乳腺组织形态学分析,采用Real-time PCR检测乳腺组织中线粒体凋亡信号通路的mRNA表达情况;采用Western blotting检测乳腺组织中磷酸酯酶与张力蛋白同源物/磷脂酰肌醇3-激酶/蛋白激酶B(PTEN/PI3K/AKT)和线粒体凋亡信号通路相关蛋白表达。结果:与正常对照组比较,模型对照组大鼠行为学评分、乳房直径、乳头高度显著增加,乳腺导管上皮细胞增生层数和乳腺组织病理学评分增加,血浆中CD^(8+)T淋巴细胞亚群、血清E2、LH、FSH、PRL和乳腺组织中Cyto C含量及E2/P、E2/T比值升高,Pcna、Bcl2、Bclxl mRNA表达和Bcl-2/Bax、Bcl-xl/Bad比值升高,pro-Caspase-3、pro-Caspase-9蛋白表达上调,p-PI3K/PI3K、p-AKT/AKT比值显著升高(P<0.01);血浆中CD^(3+)、CD^(4+)T淋巴细胞亚群、P、T含量,乳腺组织中Cyto C含量及CD^(4+)/CD^(8+)比值显著降低,Bax、Bad、Apaf1 mRNA表达下调,PTEN、cleaved-Caspase-3、cleaved-Caspase-9和cleaved-PARP-1蛋白表达显著下调(P<0.01);用乳癖散结汤34、68 g/kg治疗后,上述指标被明显逆转(P<0.05或P<0.01)。结论:乳癖散结汤对MGH大鼠具有较好的治疗作用,其作用机制可能与其增强免疫功能、调节性激素及PTEN/PI3K/AKT信号通路,从而促进线粒体途径介导的细胞凋亡密切相关。
Objective:To study the therapeutic effect and underlying mechanism of Rupi Sanjie Decoction(乳癖散结汤)on rats with mammary gland hyperplasia(MGH).Methods:Rats were randomized into normal group,model group,Rupi Sanjie Decoction(乳癖散结汤)(34,68 g/kg)groups and tamoxifen(0.0036 g/kg)group.The administration(ig)lasted 8 weeks.Then,the behavioral observation was performed and breast width diameter and nipple height were measured.The content of T lymphocyte subsets(CD^(3+),CD^(4+),CD^(8+))and sex hormones[(follicle-stimulating hormone(FSH),luteinizing hormone(LH),prolactin(PRL),estradiol(E2),testosterone(T),progestin(P)]in plasma were determined.The release of mitochondrial cytochrome C(Cyto C)from breast tissue cells and breast histomorphology were analyzed.The mRNA expression of mitochondrial apoptosis signaling pathway was detected by real-time PCR.The expression of phosphatase and tensin homologue/phosphoinositide 3-kinase/protein kinase B(PTEN/PI3 K/Akt)and proteins in mitochondrial apoptosis signaling pathway was measured by Western blotting.Results:Compared with the normal group,the model group showed increase in behavioral score,breast width diameter,nipple height,layers of mammary ductal epithelial cells,breast histopathological score,plasma level of CD^(8+),serum content of E2,LH,FSH,and PRL,E2/P and E2/T ratios,and Cyto C content in mammary tissue,mRNA expression of proliferating cell nuclear antigen(PCNA),B-cell lymphoma 2(Bcl-2),and Bcl-xl,Bcl-2/Bax and Bcl-xl/Bad ratios,protein expression of pro-Caspase-3 and pro-Caspase-9,and phosphorylated(p)-PI3 K/PI3 K and p-AKT/AKT ratios in breast tissue(P<0.01),and decrease in levels of CD^(3+)and CD^(4+),content of P and T,CD^(4+)/CD^(8+)ratio in the blood,Cyto C content in the breast tissue,mRNA expression of Bax,Bad,and Apaf1,and protein expression of PTEN,cleaved-Caspase-3,cleaved Caspase-9,and cleaved-poly(ADP-ribose)polymerase-1(PARP-1)in the breast tissue(P<0.01).After treatment with Rupi Sanjie Decoction(乳癖散结汤)(34,68 g/kg),the above indexes were significantly reversed compared with those in the model group(P<0.05 or P<0.01,respectively).Conclusion:Rupi Sanjie Decoction(乳癖散结汤)has therapeutic effect on MGH rats and the mechanism is that it enhances immune function and regulates sex hormones and PTEN/PI3 K/AKT signaling pathway,thereby promoting mitochondrial pathway-mediated apoptosis.
作者
张继红
石孟琼
陈茂华
梅和平
梅君
李浩然
谈燕清
李洁
王晓
梅大钊
Zhang Jihong;Shi Mengqiong;Chen Maohua;Mei Heping;Mei Jun;Li Haoran;Tan Yanqing;Li Jie;Wang Xiao;Mei Dazhao(Chinese Medicine Clinical Medical College of China Three Gorges University&Hubei Clinical Research Center for Functional Digestive Diseases of Traditional Chinese Medicine,Yichang 443002;Medical College of China Three Gorges University,Yichang 443002;Affiliated Hospital of Hubei Three Gorges Polytechnic,Yichang 443002;College of Biological and Pharmaceutical Sciences&Yichang Key Laboratory of Development and Utilization of Health Products with Drug Food Homology,Yichang 443002)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2022年第5期46-53,共8页
Pharmacology and Clinics of Chinese Materia Medica
基金
湖北省卫生和计划生育委员会项目[鄂卫生计生通(2017)20号]
宜昌市科学技术局项目(编号:A21-2-044)
三峡大学硕士学位论文培优基金项目(编号:2020SSPY144、2019SSPY159)
湖北省生物酵素工程研究技术研究中心项目(编号:JS2018-06)。
