摘要
旨在探讨中药复方清乳化岩煎(QD)对动物乳腺肿瘤的保护作用及机制。检测清乳化岩煎对正常细胞(HC11、RAW2647、DC2.4)的安全浓度,探究其对乳腺癌细胞(A72、CMT-U27、4T1)体外增殖的抑制作用;流式细胞术检测4T1细胞凋亡情况。将BALB/c小鼠分为空白对照组,模型组及清乳化岩煎低、中和高剂量组(100、200和400 mg/kg),除空白对照组外,其余小鼠皮下注射4T1细胞建立乳腺癌转移瘤模型,各组灌胃相应药物14 d,期间记录肿瘤体积和生长状况,检测脾脏淋巴细胞增殖情况,观察肿瘤组织病理变化,缺口末端标记(TUNEL)法检测移植瘤细胞凋亡水平,免疫组化(IHC)法检测肿瘤组织中基质金属蛋白酶9(MMP-9)与热休克蛋白70(HSP70)蛋白表达,ELISA法测定肿瘤组织和血清中白细胞介素2(IL-2)、IL-4、IL-10、肿瘤坏死因子α(TNF-α)、干扰素γ(IFN-γ)的含量。结果显示:当清乳化岩煎浓度大于或等于5 mg/mL时,对A27、4T1细胞的增殖存在显著抑制(P<0.001);浓度大于或等于10 mg/mL时,对CMT-U27细胞呈现出显著性抑制(P<0.001);不同浓度的清乳化岩煎均可诱导4T1细胞凋亡(P<0.001)。与模型组相比,清乳化岩煎能够显著减小肿瘤体积(P<0.001),促进肿瘤细胞坏死,改善肿瘤对周围组织的影响,促进脾脏淋巴细胞增殖(P<0.001),促进肿瘤组织的细胞凋亡(P<0.001),显著降低肿瘤组织中MMP-9和HSP70蛋白表达(P<0.05),清乳化岩煎组小鼠血清和肿瘤组织中IFN-γ和TNF-α水平显著上升(P<0.05),IL-10含量显著下降(P<0.05)。研究表明,清乳化岩煎可通过调控细胞凋亡和增强机体免疫来抑制乳腺癌的发展,为中药治疗动物乳腺癌提供理论依据。
This paper was to investigate the protective effect and mechanism of the Chinese traditional medicine prescription Qingruhuayan decoction(QD)on animals with breast cancer.The safe concentration of QD on normal cells(HC11,RAW2647,DC2.4)was detected,and its inhibitory effect on the proliferation of breast cancer cells(A72,CMT-U27,4T1)in vitro was explored.Flow cytometry was used to detect apoptosis in 4T1 cells.BALB/c mice were used and divided into the blank control group,the model group,and the low,medium,and high-dose groups of QD(100,200,and 400 mg/kg).Except the blank control group,all the groups were injected with 4T1 cells subcuta-neously to establish a breast cancer metastasis model.The corresponding drugs were administered to each group by gavage for 14 days.In this period,tumor volume and growth status in the animals were recorded,and the proliferation of spleen lymphocytes in them was examined.Pathological changes in the tumor tissue were observed,and the level of apoptosis in transplanted tumor cells was detected by TUNEL assay.Next,IHC was used to detect the expression of MMP-9 and HSP70 proteins in the tumor tissue.Finally,the levels of IL-2,IL-4,IL-10,TNF-αand IFN-γin the tumor tissue and serum were determined by ELISA.The results showed that QD significantly inhibited the prolifera-tion of A27 and 4T1 cells at concentrations higher than or equal to 5 mg/mL(P<0.001),and significantly inhibited CMT-U27 cells at con-centrations higher than or equal to 10 mg/mL(P<0.001).Different concentrations of QD all induced apoptosis in 4T1 cells(P<0.001).Compared with the model group,QD significantly reduced tumor volume in the animals(P<0.001),promoted tumor cell necrosis in them,improved the effect of the tumor on the surrounding tissues,and promoted the proliferation of their spleen lymphocytes(P<0.001).QD also promoted apoptosis in tumor tissue of the animals(P<0.001)and significantly reduced the expression of MMP-9 and HSP70 proteins in their tumor tissue(P<0.05).The levels of IFN-γand TNF-αin the serum and tumor tissue of the mice in the QD group were significantly raised(P<0.05),while the levels of IL-10 were significantly reduced(P<0.05).The present study indicated that QD inhibited the devel-opment of breast cancer by regulating cell apoptosis and enhancing the immune system,which provided a theoretical basis for the treatment of animal breast cancer with traditional Chinese medicine.
作者
罗朝婧
闫普普
万鑫
韩一恒
周辉
吴昱
李梦
郭大伟
孙文静
郭利伟
武毅
LUO Zhaojing;YAN Pupu;WAN Xin;HAN Yiheng;ZHOU Hui;WU Yu;LI Meng;GUO Dawei;SUN Wenjing;GUO Liwei;WU Yi(College of Animal Science and Technology,Yangtze University,Jingzhou 434023,China;College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China;Jiangxi Wildlife and Plant Protection Center,Nanchang 330006,China;Changzhou Affiliated Hospital of Nanjing University of Chinese Medicine,Changzhou 213000,China;College of Biology and Pharmacy,Yulin Normal University/Guangxi Key Laboratory of Agricultural Resource Chemistry and Biotechnology,Yulin 537000,China)
出处
《畜牧与兽医》
CAS
北大核心
2024年第5期127-137,共11页
Animal Husbandry & Veterinary Medicine
基金
湖北省重点研发计划项目(2023BBB045)。
关键词
清乳化岩煎
乳腺癌
凋亡
免疫增强
Qingruhuayan decoction
breast cancer
apoptosis
immune enhancement
作者简介
第一作者:罗朝婧,女,硕士研究生;通信作者:郭利伟,博士,教授,研究方向为中兽医药学,E-mail:guolw@yangtzeu edu cn;通信作者:武毅,博士,副教授,研究方向为中兽医药学,E-mail:wuyi2001cn@163 com。
