蛋白质组学分析清热解毒法作用于肝癌大鼠的血清蛋白差异表达研究  

Effects of Heat-Clearing and Detoxifying Method on Expressions of Serum Differential Proteins in Liver Cancer Rats by Proteomic Analysis

作　　者：饶希午 沈卫星[1,2] 陈婷婷 孙东东[1,2] 苗筠杰 李柳[1,2] 孙超 许宏钦 程海波[1,2] RAO Xiwu;SHEN Weixing;CHEN Tingting;SUN Dongdong;MIAO Yunjie;LI Liu;SUN Chao;XU Hongqin;CHENG Haibo(The First Clinical Medical College of Nanjing University of Chinese Medicine,Nanjing 210023,Jiangsu,China;Jiangsu Collaborative Innovation Center of Traditional Chinese Medicine Prevention and Treatment of Tumor,Nanjing 210023,Jiangsu,China;Shuguang Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200120,Jiangsu,China)

机构地区：[1]南京中医药大学第一临床医学院,江苏南京210023 [2]江苏省中医药防治肿瘤协同创新中心,江苏南京210023 [3]上海中医药大学附属曙光医院肿瘤科,上海200120

出　　处：《中华中医药学刊》2020年第4期51-56,I0012,I0013,共8页Chinese Archives of Traditional Chinese Medicine

基　　金：国家重点研发计划(2017YFC1700602);国家自然科学基金面上项目(81573910,81673559);江苏省高等自然科学研究重大项目(17KJA360002);江苏省自然科学基金面上项目(BK20141467);江苏高校优势学科建设工程项目(PAPD);江苏省研究生科研创新计划[KYCX18(1542)]。

摘　　要：目的观察清热解毒法-白花蛇舌草和半枝莲对肝癌大鼠血清蛋白差异表达的影响。方法 40只SD雄性大鼠随机分为正常组(n=10)、模型组(n=10)、清热解毒常规剂量组(n=10)、清热解毒超超高剂量组(n=10)。给予模型组与清热解毒常规剂量组、清热解毒超超高剂量组大鼠0.01%DEN(Sigma,纯度为99.9%,蒸馏水配制)溶液自由饮用8周后,停药2周,期间改为普通喂养,继续给药喂养6周。清热解毒常规剂量组与清热解毒超超高剂量组大鼠于第4周分别给予剂量为4.98、29.88 g·kg-1·d-1的白花蛇舌草、半枝莲水提液灌胃,每周6 d。模型组于第4周同时给予0.9%生理盐水灌胃,1 mL/d,每周6 d。大鼠于第16周灌胃结束后全部处死,取血清样本。将3组血清样本进行TMT肽段标记,质谱鉴定,应用KOBAS软件分析3组差异表达蛋白质的生物信息学。结果共鉴定到1151个蛋白点。模型组与正常组相比,差异表达明显的蛋白共234个,包含上调蛋白151个,下调蛋白83个。清热解毒常规剂量组与模型组相比,差异表达明显的蛋白共216个,包括上调蛋白100个,下调蛋白116个。清热解毒超超高剂量组与模型组相比,差异表达明显的蛋白共208个,包括上调蛋白84个,下调蛋白124个。清热解毒超超高剂量组与清热解毒常规剂量组相比,差异表达明显的蛋白共40个,包括上调蛋白27个,下调蛋白13个;对所有差异表达蛋白对应基因于PUBMED数据库逐一进行检索,发现这些差异表达蛋白主要参与了代谢、炎症反应、免疫调控、自噬、热休克、氧化应激、细胞周期与凋亡、细胞生长与分化、基质降解、血管生成、肿瘤转移、癌相关、肝再生修复等。信号通路分析涉及谷胱甘肽代谢、PPAR代谢等代谢相关通路,T细胞、B细胞、NK细胞激活相关通路,血管新生相关如VEGF信号通路,细胞生长与分化如TGF-β相关通路,Ras、P38 MAPK、Wnt等通路。结论研究证实清热�Objective To observe the effects of heat-clearing and detoxification methods(QRJD)-Baihuasheshecao(Hedyotis diffusa) and Banzhilian(Scutellaria barbata) on the differential expressions of serum proteins in liver cancer rats. Methods Forty male SD rats were randomly divided into normal group(n=10), model group(n=10), QRJD low dose group(n=10), and QRJD high dose group(n=10). Rats in the model group and the QRJD low and high dose groups were supplied with 0.01% DEN solution(Sigma, purity of 99.9%, distilled water) for free drinking for eight weeks, then we set up a discontinuous period for 2 weeks and restarted the 0.01% DEN solution for free drinking for 6 weeks. The QRJD low and high dose groups received water extract from Baihuasheshecao(Hedyotis diffusa) and Banzhilian(Scutellaria barbata)[4.98 g·(kg-1·d-1) and 29.88 g·(kg-1·d-1)] by gavage administration respectively from the 4 th week to the 16 th week while the model group received 0.09% normal saline(1 mL·d-1),6 days per week. All rats were executed at the end of the 16 th week, and the serum samples were taken out. The identification was performed by tandem mass tags(TMTs) and mass spectrometry and the results were processed by using KOBAS software for bioinformatics analysis. The protein expression before and after the drug action was observed and compared. Results A total of 1151 proteins were identified. Compared with the normal group, there were 234 differentially expressed proteins in the model group, including 151 up-regulated proteins and 83 down-regulated proteins. Compared with the model group, there were 216 differentially expressed proteins in the QRJD low dose group, including 100 up-regulated proteins and 116 down-regulated proteins. In the QRJD high dose group, there were 208 differentially expressed proteins, including 84 up-regulated proteins and 124 down-regulated proteins. Compared with QRJD low dose group,there were 40 differentially expressed proteins in the QRJD high dose group, including 27 up-regulated proteins and 13 down-regul

关 键 词：肝癌 清热解毒 蛋白组学 

分 类 号：R285.5[医药卫生—中药学]