张帅,朱晓青,王艳丽,程枝梅,李勇军,陆苑
ZHANG Shuai,ZHU Xiaoqin,WANG Yanli,CHENG Zhimei,LI Yongjun,LU Yuan

• 1:贵州医科大学附属医院介入科
• 2:贵州医科大学药学院
• 3:贵州医科大学民族药与中药开发应用教育部工程研究中心
• 4:贵州医科大学贵州省药物制剂重点实验室/省部共建药用植物功效与利用国家重点实验室

摘要(Abstract)：

目的 观察吡非尼酮（PFD）联合阿霉素（DOX）对小鼠H22肝癌移植瘤的协同抗肿瘤作用并探讨可能的作用机制。方法 雄性H22移植瘤模型小鼠40只随机均分为4组，对照组（生理盐水0.25 mL/d）、PFD+DOX末次组[PFD 200 mg/（kg·d），最后一天给DOX]、DOX组（DOX 2.5 mg/kg）、PFD+多次DOX组[PFD200 mg/（kg·d），隔日在PFD 200 mg/kg给药1 h后给予DOX 2.5 mg/kg]，连续15 d；给药周期内隔天称量体重，测量瘤体体积并绘制肿瘤的生长曲线；试验结束后处死小鼠，剥离肿瘤组织并称重，计算抑瘤率和PFD+DOX联用相互作用指数Q值，HE染色法观察各组小鼠H22实体瘤细胞形态，UPLC-MS/MS法测定各组小鼠血浆、肿瘤、心、肝、肾组织中的DOX和阿霉素醇（DOXol）的浓度。结果 PFD+DOX联用Q值为1.47，提示PFD与DOX联用对抑制肿瘤的生长有协同作用；PFD+多次DOX组与DOX组比较，脾脏指数显著上升（P<0.01）；与对照组比较，PFD+多次DOX组的肿瘤组织出现大面积空隙，呈现片状坏死或消融现象比DOX组更加明显；与PFD+末次DOX组相比，DOX组血清及各组织中DOX的累积含量显著升高（P<0.01），肿瘤、心、肝、肾及肺组织中DOXol的累积含量显著升高（P<0.01），但在血清中未测到DOXol；与DOX组相比，PFD+多次DOX组肿瘤中DOX和DOXol的浓度增加（P<0.05），可降低心脏组织中DOXol的浓度（P<0.05）。结论 PFD与DOX联合应用，对H22移植瘤小鼠有抗肿瘤协同作用，其机制可能与PFD增加肿瘤组织中DOX及DOXol浓度有关。
Objective To observe the synergistic antitumor effect of pirfenidone(PFD) combined with doxorubicin( DOX) on H22 hepatocellular carcinoma( HCC)-bearing mouse model, and to explore its possible mechanism.Methods H22 HCC-bearing mouse model was established. The model mice were randomly divided into 4 groups(10 mice/group): control group( normal saline,0. 25 mL/d), PFD group [PFD, 200 mg/（kg·d）], DOX group [DOX, 2. 5 mg/（kg·d）], PFD +DOX group [ PFD 200 mg/（ kg · d ） + DOX 2. 5 mg/kg/every other day) at 1 h after PFD administration] for 15 consecutive days. During the administration, mouse body weights were measured every other day. The long diameters(a) and short diameters(b) of the mouse tumors were measured with vernier calipers. The tumor volumes were calculated and the tumor growth curves were drawn. The PFD group, DOX group and PFD + DOX group were given DOX during the administration.After the experiment, the mice were sacrificed, and the tumors was collected and weighed. The tumor inhibition rates and the PFD-DOX interaction index Q values were calculated. HE staining was used to observe the morphology of H22 HCC cells in each group. The concentrations of DOX and doxorubicin alcohol(DOXol) in plasma, tumor, heart, liver, and kidney in each group were determined using UPLC-MS/MS.Results The Q value of PFD + DOX was 1. 47, indicating that PFD and DOX may have a synergistic effect in inhibiting tumor growth. Compared with DOX group, the spleen index of the DOX + PFD group was significantly increased(P< 0. 01). When compared to control group, the tumors in DOX + PFD group had a large area of voids, showing sheet-like necrosis or ablation, which was more obvious than that in DOX group. Compared with PFD group(single administration of DOX),the cumulative contents of DOX and its metabolite DOXol in the serum and various tissues in DOX group(multiple administration of DOX) were significantly increased(P< 0. 01). When compared to DOX group, the concentrations of DOX and DOXol were increased(P< 0. 05), but the concentrations of DOXol were decreased in the hearts in PFD + DOX group(P< 0. 05).Conclusion PFD may exert a synergistic anti-tumor effect with DOX by increasing the concentrations of DOX and DOXol in tumors.

关键词(KeyWords)： 吡非尼酮;阿霉素;抗肿瘤作用;协同作用;组织分布
pirfenidone(PFD);doxorubicin;antitumor effect;synergistic effect;biodistribution

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金资助项目（81860320）;; 贵州省高层次留学人才创新创业重点项目（留学人才择优资助合同[2019]01）;; 贵州省自然科学基金项目（黔科合基础[2018]1129）;; 贵州医科大学附属肿瘤医院院内项目（YJ2019018）;贵州医科大学附属医院博士启动基金

作者(Author): 张帅,朱晓青,王艳丽,程枝梅,李勇军,陆苑
ZHANG Shuai,ZHU Xiaoqin,WANG Yanli,CHENG Zhimei,LI Yongjun,LU Yuan

DOI: 10.19367/j.cnki.2096-8388.2022.03.002

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