白蔹乙酸乙酯部位体内抗肿瘤的药效作用机制及其成分分析研究  

Study on the Mechanism of Antitumor Effect of Ethyl Acetate Extract of Ampelopsis Radix In Vivo and Its Composition Analysis

作　　者：韩思婕 潘翔 张梦美[1] 魏琼 叶晓川[1] HAN Sijie;PAN Xiang;ZHANG Mengmei;WEI Qiong;YE Xiaochuan(Hubei University of Chinese Medi Provincial Key Laboratory of Chinese Medicine Hubei Resource and Chemistry,Wuhan 430065 Hubei,China)

机构地区：[1]湖北中医药大学,中药资源与中药化学湖北省重点实验室,湖北武汉430065

出　　处：《中药新药与临床药理》2022年第12期1623-1631,共9页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：湖北省技术创新专项重点项目(2019AFB866)。

摘　　要：目的 探究白蔹乙酸乙酯部位对H22实体瘤的抑制作用及机制,以及对正常小鼠的急性毒性,并分析其药效物质基础。方法 对40只KM小鼠皮下注射H22细胞悬液复制H22皮下移植瘤小鼠模型,随机分为模型组、氟尿嘧啶组(5-Fu,25 mg·kg^(-1))和白蔹乙酸乙酯部位低、中、高(1.5、4.5、13.5 g·kg^(-1),生药量)剂量组,每组8只。白蔹乙酸乙酯部位灌胃给药,每日1次,连续10 d。测定肿瘤体积、质量,计算抑瘤率;计算肝脏、脾脏、胸腺的脏器指数;采用ELISA法检测血清中肿瘤坏死因子α(TNF-α)、白细胞介素2(IL-2)和白细胞介素6(IL-6)的水平;HE染色法观察肿瘤组织病理变化;Western Blot法测定肿瘤组织中Bax、Bcl-2、Caspase-3、Caspase-9蛋白表达水平。以最大给药剂量灌胃(30 mL·kg^(-1))1次并连续观察14 d,考察白蔹水提物、醇提物、乙酸乙酯部位(4.00、5.14、23.22 g·mL(-1))对正常小鼠的急性毒性作用。采用超高效液相色谱-四极杆-飞行时间串联质谱(UPLC-QTOF-MS/MS)法分析白蔹乙酸乙酯部位的化学成分。结果 与模型组相比,白蔹乙酸乙酯部位高剂量组的肿瘤体积明显减小(P<0.05),各给药组的肿瘤质量均显著降低(P<0.01),低、中、高剂量组的抑瘤率分别为64.42%、72.00%、78.24%,肝脏、脾脏、胸腺的脏器指数均明显升高(P<0.05,P<0.01);低剂量组小鼠的血清TNF-α、IL-2、IL-6水平均显著升高(P<0.01),高剂量组小鼠的血清IL-6水平显著升高(P<0.01);各给药组小鼠肿瘤组织中Bcl-2蛋白表达均显著下调(P<0.01),Bax、Caspase-3及Caspase-9蛋白表达均明显上调(P<0.05,P<0.01);白蔹乙酸乙酯部位高剂量组的Bax/Bcl-2蛋白比值显著升高(P<0.01)。小鼠在给药剂量相当于生药量696.6 g·kg^(-1)·d-1(约相当于临床使用最大剂量的462.8倍)的白蔹乙酸乙酯部位时,无明显毒副反应。UPLC-QTOF-MS/MS分析共鉴定出16个化学成分,以酚酸类为主。结论 白蔹乙酸乙酯部位具有明Objective To investigate the inhibitory effect and mechanism of the ethyl acetate extract of Ampelopsis Radix on H22 solid tumours and its acute toxicity to normal mice, and to analyze its pharmacological basis.Methods Forty KM mice were injected subcutaneously with H22 cells suspension to replicate the H22 solid transplant tumor mouse model,and randomly divided into the model group,the fluorouracil group(5-Fu,25 mg·kg^(-1)) and low-, medium-and high-(1.5, 4.5 and 13.5 g · kg-1, raw dose) dose of ethyl acetate extract of Ampelopsis Radix groups,with 8 mice in each group. The ethyl acetate extract administration groups were given intragastric administration once a day for consecutive 10 days. The tumor volume and mass were measured and the tumor suppression rate was calculated;the organ indices of liver,spleen and thymus were calculated;the serum levels of tumor necrosis factor α(TNF-α),interleukin 2(IL-2)and interleukin 6(IL-6)were detected by ELISA;the tumor histopathological changes were observed by HE staining. The protein expression levels of Bax,Bcl-2,Caspase-3 and Caspase-9 in the tumour tissue were determined by Western Blot. The acute toxic effects of the aqueous extract,alcoholic extract and ethyl acetate site(4.00,5.14 and 23.22 g·mL^(-1))of Ampelopsis Radix were investigated in normal mice by gavage(30 mL · kg-1) at the maximum administered dose once and continuously observed for 14 days. The chemical composition of the ethyl acetate extract of Ampelopsis Radix was analyzed by ultra performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry(UPLC-QTOF-MS/MS). Results Compared with the model group,the tumor volume was significantly reduced in the high-dose group of ethyl acetate extract of Ampelopsis Radix group(P<0.05),and the tumor mass was significantly reduced in all dosing groups(P<0.01);the tumor inhibition rates were 64.42%,72.00% and 78.24% in the low-,medium-and high-dose groups,respectively,and the organ indexes of liver,spleen and thymus were significantly incre

关 键 词：白蔹 乙酸乙酯部位 H22荷瘤小鼠 急性毒性 细胞凋亡 Bax/Bcl-2/Caspase-9/Caspase-3途径 酚酸类 超高效液相色谱-四极杆-飞行时间串联质谱 

分 类 号：R285.5[医药卫生—中药学]