摘要
目的:探讨香草扶正合剂(XCFZ)对Lewis肺癌小鼠的抗肿瘤效应机制。方法:采用Lewis肺癌细胞株培养移植法建立肺癌荷瘤小鼠模型,随机分为模型组,顺铂组,XCFZ低、中、高剂量组,联合组,每组10只;造模第2天开始,XCFZ低、中、高剂量组分别给予XCFZ 0.93、1.86、3.72 g/mL灌胃;顺铂组以0.64 mg/mL顺铂溶液0.1 mL/2 d腹腔注射,同时以双蒸水0.2 mL/d灌胃,联合组以XCFZ中剂量灌胃和顺铂腹腔注射联合给药,模型组按照等量0.9%氯化钠溶液给药,每天1次,连续给药14d;称取小鼠体质量、瘤重、脾脏及胸腺重量,并计算抑瘤率、脾指数、胸腺指数,HE染色观察肿瘤组织形态学变化,qRT-PCR检测肿瘤组织中HK2、LDH-A mRNA表达,免疫组化及Western Blot检测瘤组织VEGFR-2、VEGF、PI3K、Akt蛋白的表达。结果:联合组抑瘤率最高,其次为顺铂组和XCFZ中剂量组。与模型组比较,XCFZ高、中剂量组及联合组脾指数显著升高(P<0.01,P<0.05),XCFZ高、中剂量组胸腺指数显著升高(P<0.01,P<0.05),顺铂组胸腺指数及脾指数显著降低(P<0.01,P<0.05);与顺铂组比较,XCFZ各剂量组及联合组胸腺指数显著升高(P<0.05,P<0.01),XCFZ中、高剂量组和联合组脾指数显著升高(P<0.01)。XCFZ各用药组肿瘤细胞核仁固缩,细胞坏死和核碎裂现象亦较多;顺铂组肿瘤细胞较小,核分裂现象和异型细胞少见,坏死碎裂细胞较多。与模型组比较,XCFZ各剂量组、顺铂组和联合组LDH-A和HK2 mRNA表达水平显著降低(P<0.05,P<0.01);与顺铂组比较,联合组LDH-A mRNA表达水平显著降低(P<0.05)。与模型组比较,XCFZ中、高剂量组及顺铂组、联合组小鼠肿瘤组织中VEGFR-2、VEGF、PI3K、Akt蛋白表达均显著降低(P<0.01,P<0.05);与顺铂组比较,联合组VEGFR-2、VEGF、PI3K、Akt蛋白表达均显著降低(P<0.05,P<0.01)。结论:XCFZ联合顺铂对Lewis肺癌小鼠有改善免疫功能及增效减毒作用,其机制是通过PI3K/Akt通路干预VEGF/VEGFR-2抗肿瘤血管生成,以及下调糖酵解LDH-A基因表达来抑制肺癌增殖转移。
Objective:To investigate the mechanism of anti-tumor effect of Xiangcao Fuzheng Mixture(XCFZ)on Lewis lung cancer mice.Methods:Lewis lung cancer cell line culture and transplantation method was used to establish a lung cancer tumor bearing mouse model,which was randomly divided into model group(M),chemotherapy group(DDP),XCFZ low(LD),medium(MD),high dose group(HD),and combined group(MD+DDP),with 10 mice in each group.From the second day of modeling,the LD,MD,HD were gavaged XCFZ 0.93,1.86 and 3.72 g/mL,respectively.DDP was intraperitoneally injected with 0.64 mg/mL cisplatin solution 0.1 mL/2 d,and was intragastatically administrated with 0.2 mL/d double steaming water.The MD+DDP was administrated with XCFZ medium dose intragastrium combined with intraperitoneal injection of cisplatin.The model group was administrated with 0.9%sodium chloride solution once a day for 14 days.The body weight,tumor weight,spleen and thymus weight of the mice were weighed,and the tumor inhibitory rate and spleen thymus index were calculated.The mRNA expressions of HK2 and LDH-2 in tumor tissues were detected by qRT-PCR,and the expressions of VEGFR-2,VEGF,PI3K and Akt in tumor tissues were detected by immunohistochemistry and Western Blot.Results:The tumor inhibition rate of MD+DDP group was the highest,followed by DDP group and MD group.Compared with M group,spleen index in XCFZ highdose and medium-dose groups and MD+DDP groups was significantly increased(P<0.01,P<0.05),thymus index in XCFZ highdose and medium-dose groups was significantly increased(P<0.01,P<0.05),spleen index and thymus index in DDP group was significantly decreased(P<0.01,P<0.05).Compared with the DDP group,the thymus index of all XCFZ dose groups and the MD+DDP group was significantly increased(P<0.05,P<0.01),and the spleen index of XCFZ middle and high dose groups and the MD+DDP group was significantly increased(P<0.01).XCFZ treatment groups had more nucleolar shrinkage,cell death and nuclear fragmentation.In the cisplatin group,the tumor cells were smaller,the nuclear division phenomenon and heterotypic cells were rare,and the necroclastic cells were more.Compared with the M group,the expression levels of LDH-A and HK2 mRNA in each dose of XCFZ group,cisplatin group and combination group were significantly decreased(P<0.05,P<0.01).Compared with the DDP group,the expression level of LDH-A mRNA in the combination group was significantly decreased(P<0.05).Compared with the M group,the expressions of VEGFR-2,VEGF,PI3K and Akt protein in tumor tissues of XCFZ medium and high dose groups,DDP group and MD+DDP group were significantly decreased(P<0.01,P<0.05).Compared with the DDP group,the expressions of VEGFR-2,VEGF,PI3K and Akt protein in the combination group were significantly decreased(P<0.05,P<0.01).Conclusion:XCFZ combined with Cisplatin can improve immune function.enhance efficacy and reduce toxicity in Lewis lung cancer mice.The mechanism is to interfere with VEGF/VEGFR-2 anti-tumor angiogenesis through PI3K/Akt pathway,and down-regulate the expression of glycolytic LDH-A gene 10 inhibit the proliferation and metastasis of lung cancer.
作者
程权
王硕
傅华洲
CHENG Quan;WANG Shuo;FU Huazhou(Affiliated Hangzhou First People's Hospital,School of Medicine,Westlake University,Hangzhou 310006,China;Zhejiang Xiaoshan Hospital,Hangzhou 311202,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2024年第7期3717-3723,共7页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
国家中医药管理局第七批全国老中医药专家学术经验继承工作室建设项目(No.国中医药人教函[2022]76号)
浙江省中医药防治重大疾病攻关计划项目(No.2018ZY009)
浙江省名老中医专家传承工作室建设项目(No.GZS2020029)。
作者简介
通信作者:程权,浙江省杭州市上城区浣纱路261号西湖大学医学院附属杭州市第一人民医院,邮编:310006,E-mail:hzallanqcheng@163.com。
