复方蜥蜴散调控PI3K/AKT/MMP通路对胃癌耐药细胞增殖、迁移及侵袭的影响  

Compound Lizard Powder Regulates the PI3K/AKT/MMP Pathway to Inhibit the Proliferation,Migration,and Invasion of Cisplatin-Resistant Gastric Cancer Cells

作　　者：李铮 程翻娥[3] 刘彩月 朱西杰 师小茜[1,2] 李卫强 LI Zheng;CHENG Fan'e;LIU Caiyue;ZHU Xijie;SHI Xiaoqian;LI Weiqiang(School of Traditional Chinese Medicine,Ningxia Medical University,Yinchuan 750004;Key Laboratory of Ningxia Ethnomedicine Modernization,Ministry of Education,Yinchuan 750004;Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100700)

机构地区：[1]宁夏医科大学中医学院,银川750004 [2]宁夏少数民族医药现代化教育部重点实验室,银川750004 [3]北京中医药大学东直门医院,北京100700

出　　处：《中药药理与临床》2025年第1期45-51,共7页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金项目(编号:82260916);宁夏医科大学科学研究资助项目(编号:XZ2023012);宁夏自然科学基金重点项目(编号:2021AAC02014)。

摘　　要：目的:观察复方蜥蜴散通过介导信号通路磷脂酰肌醇-3激酶(PI3K)/蛋白激酶B(AKT)/基质金属蛋白酶(MMP)对胃癌化疗耐药MKN45/DDP细胞增殖、迁移及侵袭和耐药相关蛋白表达的影响。方法:40只SD大鼠随机分为正常对照组、复方蜥蜴散17.2、34.4、68.8 g/kg组,每组10只,制备含药血清。CCK-8法检测复方蜥蜴散17.2、34.4、68.8 g/kg单用及各组与顺铂联合用药对MKN45/DDP细胞增殖的影响,测得68.8 g/kg复方蜥蜴散20%含药血清单用及与顺铂联合用药的抑制作用最佳,用于进行后续实验。后续细胞实验中分为空白对照组、顺铂1.2μg/mL组、复方蜥蜴散20%含药血清组、复方蜥蜴散20%含药血清联合顺铂1.2μg/mL组、复方蜥蜴散20%含药血清+激动剂740Y-P 10μmol/L组。采用细胞克隆形成实验、划痕实验及Transwell实验检测各给药组对MKN45/DDP细胞增殖、迁移及侵袭能力的影响;采用Western blot法检测各组细胞中PI3K/AKT/MMP信号通路相关蛋白以及耐药相关蛋白多药耐药蛋白6(MRP6)、P-糖蛋白(P-gp)的表达。结果:17.2、34.4、68.8 g/kg复方蜥蜴散20%含药血清组及与顺铂联合用药组均能明显抑制细胞增殖(P<0.05),并呈现剂量依赖性,其抑制作用随剂量升高而增强。与空白对照组和顺铂1.2μg/mL组相比,68.8 g/kg复方蜥蜴散20%含药血清组、68.8 g/kg复方蜥蜴散20%含药血清+顺铂1.2μg/mL组及68.8 g/kg复方蜥蜴散20%含药血清+740Y-P 10μmol/L组均能抑制MKN45/DDP细胞的增殖、迁移及侵袭(P<0.05或P<0.01),且明显下调MMP-2、MMP-9、Vimentin、P-PI3K、P-AKT蛋白表达(P<0.05或P<0.01),上调E-cadherin表达(P<0.01);与复方蜥蜴散20%含药血清组相比,复方蜥蜴散20%含药血清+顺铂1.2μg/mL组明显抑制细胞增殖、迁移及侵袭(P<0.05或P<0.01),而复方蜥蜴散20%含药血清+740Y-P 10μmol/L组抑制能力明显减弱(P<0.05或P<0.01),且MMP-2、MMP-9、Vimentin、P-PI3K、P-AKT、MRP6、P-gp蛋白表达明显上调(P<0.05Objective: To observe the effects of Compound Lizard Powder on the proliferation, migration, invasion, and expression of resistance-related proteins of cisplatin-resistant MKN45/DDP cells via the phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/matrix metalloproteinase(MMP) signaling pathway. Methods: Forty SD rats were randomized into blank control and Compound Lizard Powder(17.2, 34.4, and 68.8 g/kg) groups, with 10 rats in each group. The sera containing different doses of Compound Lizard Powder were then prepared. The CCK-8 method was used to detect the effects of Compound Lizard Powder(17.2, 34.4, and 68.8 g/kg)-containing sera in single use and in combination with cisplatin on the proliferation of MKN45/DDP cells. The results showed that the serum containing Compound Lizard Powder administrated at 68.8 g/kg in single use and in combination with cisplatin had the best inhibitory effects, and thus it was used for the subsequent experiment. The subsequent experiment was designed with the following groups: blank control, cisplatin(1.2 μg/mL), 20% serum containing Compound Lizard Powder, 20% serum containing Compound Lizard Powder + cisplatin(1.2 μg/mL), and 20% serum containing Compound Lizard Powder + agonist 740Y-P(10 μmol/L). The proliferation, migration, and invasion of MKN45/DDP cells in each group were examined by the colony formation assay, scratch assay, and Transwell assay, respectively. Western blotting was employed to determine the expression levels of proteins in the PI3K/AKT/MMP signaling pathway, as well as resistance-related proteins such as multidrug resistance protein 6(MRP6) and P-glycoprotein(P-gp) in each group of cells. Results: The 20% sera containing Compound Lizard Powder administrated at 17.2, 34.4, and 68.8 g/kg, as well as the combined therapy with cisplatin inhibited cell proliferation(P<0.05) in a dose-dependent manner. Compared with the blank control group and the cisplatin(1.2 μg/mL) group, the 20% serum containing Compound Lizard Powder, 20% serum containing Compound

关 键 词：复方蜥蜴散 胃癌顺铂耐药 增殖 迁移 侵袭 磷脂酰肌醇-3激酶 蛋白激酶 金属蛋白酶 

分 类 号：R285[医药卫生—中药学]