目的:探讨苓桂术甘汤联合基质金属蛋白酶(matrix metalloproteinases,MMP)响应型多柔比星(doxorubicin,DOX)逐级靶向脂质体抗原发性肝癌的协同增效作用.方法:以HepG2细胞为体外细胞模型,利用在线SynergyFinder软件分析苓桂术甘汤联合MMP酶响应型DOX逐级靶向脂质体的协同指数;50只3周龄SPF级C57BL/6小鼠随机分为2组,空白对照组(n=10)和造模组(n=40).采用腹腔注射二乙基亚硝胺(diethylnitrosamine,DEN)和高脂饮食相结合建立原发性肝癌小鼠模型,模型制备成功后将小鼠分为模型组(Model)、苓桂术甘汤组(LGZGT)、脂质体组(Lip)及联合干预组(LGZGT+Lip),各治疗组小鼠分别灌胃给予苓桂术甘汤,腹腔注射给予脂质体,其余各组予等量生理盐水,连续8周.考察小鼠体重、肝脏指数;甲胎蛋白(alpha-fetoprotein,AFP)、癌胚抗原(carcinoembryonic antigen,CEA)水平,谷草转氨酶(aspartate ami-notransferase,AST)、谷丙转氨酶(alanine aminotransferase,ALT)、γ-谷氨酰转移酶(γ-glutamyl transferase,γ-GT)、碱性磷酸酶(alkaline phosphatase,AKP)活性;苏木素伊红(hematoxylin eosin,HE)染色、马松(Masson)染色观察肝脏病理变化;免疫组织化学染色、原位末端标记法(terminal-deoxynucleoitidyl transferase mediated nick end labeling,TUNEL)染色观察肿瘤细胞增殖、凋亡情况.结果:两药联合协同指数评分结果显示,两药联合应用零相互作用效力(zero interaction potency,ZIP)值为2.697,具有协同增效作用.体内药效学结果显示,苓桂术甘汤,脂质体及联合干预后,均能降低小鼠肝脏指数、AFP和CEA表达水平以及ALT,AKP,AST,γ-GT活性,改善肝脏病理损伤、肝纤维化,抑制肝癌细胞增殖及促进肝癌细胞凋亡,且LGZGT+Lip组的干预能力最强.结论:苓桂术甘汤与MMP响应型DOX逐级靶向脂质体联合应用于原发性肝癌的治疗,具有协同增效作用,为临床原发性肝癌的治疗提供了新思路.
Synergistic effect of Linggui Zhugan Decoction combined with matrix metalloproteinases enzyme-responsive doxorubicin stepwise targeting liposomes on primary liver cancer
Objective:To investigate the synergistic effect of Linggui Zhugan Decoction combined with matrix metalloproteinases(MMP)and doxorubicin(DOX)targeting liposomes on primary liver cancer.Methods:HepG2 cells were used to build cell model in vitro.Synergistic index of Linggui Zhugan Decoction combined with MMP enzyme-responsive DOX targeting liposomes was analyzed by SynergyFinder software.Fifty 3-week-old SPF C57BL/6 mice were randomly divided into 2 groups:blank control group(n=10)and modeling group(n=40).A mouse model of primary liver cancer was established by intraperitoneal injection of diethylnitrosamine(DEN)combined with a high-fat diet.Subsequently,the mice were divided into model group,Linggui Zhugan group(LGZGT),liposomes group(Lip)and combined treatment group(LGZGT+Lip).Mice in each treatment group were given LGZGT by gavage,Lip by intraperitoneal injection,and the mice in the other groups were given equal amount of normal saline for 8 weeks.The body weight and liver index of mice were recorded.The levels of alpha-fetoprotein(AFP)and carcinoembryonic antigen(CEA),the activities of aspartate aminotransferase(AST),alanine aminotransferase(ALT),γ-glutamyl transferase(γ-GT),and alkaline phosphatase(AKP),the pathological changes of liver were observed by HE staining and Masson staining.The proliferation and apoptosis of tumor cells were observed by immunohistochemical staining and TUNEL staining.Results:The results of synergistic index showed that the ZIP value of the two drugs combined intervention was 2.697,indicating synergistic effect.In vivo pharmacodynamic results showed that LGZGT,Lip and LGZGT+Lip intervention can reduce liver index,expression levels of AFP and CEA,activities of ALT,AKP,AST,and γ-GT,ameliorate liver pathological injury and liver fibrosis,inhibit the proliferation of liver cancer cells and promote apoptosis of liver cancer cells.It turns out that LGZGT+Lip has the strongest intervention ability.Conclusion:Linggui Zhugan Decoction combined with MMP-responsive DOX stepwise targeting liposomes in the treatment of primary liver cancer has a synergistic effect,providing a new idea for the clinical treatment of primary liver cancer.
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