基于能量代谢探讨黄芪颗粒对H22荷瘤小鼠化疗增效的作用机制  

Mechanism of Astragalus Granule Enhancing the Efficacy of Chemotherapy in H22 Tumor-bearing Mice Based on Energy Metabolism

作　　者：李影[1,2] 邓晓霞 李清宋 朱秋菊 刘红宁 林色奇[1,2] LI Ying;DENG Xiaoxia;LI Qingsong;ZHU Qiuju;LIU Hongning;LIN Seqi(Research Center forDifferentiation and Development of TCM Basic Theory,Jiangxi University of Traditional Chinese Medicine,Nanchang 330004 Jiangxi,China;Jiangxi Province Key Laboratory of TCM Etiopathogenisis,Nanchang 330004 Jiangxi,China;Jiangxi Health Vocational College in Jiangxi Province,Nanchang 330052 Jiangxi,China)

机构地区：[1]江西中医药大学中医基础理论分化发展中心,江西南昌330004 [2]江西省中医病因生物学重点实验室,江西南昌330004 [3]江西卫生职业学院,江西南昌330052

出　　处：《中药新药与临床药理》2019年第10期1183-1188,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目（81160493,81660738）;江西省教育厅科学技术研究项目（KJ201209263065）;江西中医药大学省级重点学科建设经费资助重点重大科研项目（2013jzzdxk066）;江西省2017年度研究生创新专项基金项目（YC-2017-S354）

摘　　要：目的基于能量代谢探讨黄芪颗粒对H22荷瘤小鼠化疗增效的作用机制。方法将18只雄性KM小鼠随机分成模型组(M组)、氟尿嘧啶组(5FU组,25 mg·kg-1·d-1)、黄芪颗粒组(H组,2.8 g·kg-1·d-1)。于小鼠右侧前肢腋窝皮下注射0.2 mL(1.5×107·mL-1)的H22细胞悬液,建立肝癌小鼠模型。5FU组和H组于造模次日开始给予5FU腹腔注射,每日1次,连续7 d;第8天开始,H组给予黄芪颗粒灌胃给药,每日1次,连续7d。采用HE染色观察肿瘤细胞的侵袭程度;免疫组化法检测肿瘤组织中CD68、CD163、白细胞介素-6(interleukin-6,IL-6)阳性细胞的表达情况;实时荧光定量PCR法检测肿瘤组织中磷酸甘油酸激酶1(phosphoglycerate kinase 1,PGK1)、缺氧诱导因子-1α(hypoxiain-duciblefactors-1alpha, HIF-1α)mRNA表达情况。结果与M组比较,5FU组小鼠的肿瘤体积显著减小(P <0.01);对周围组织的侵袭、炎性细胞浸润、新生血管及核分裂象均明显减少;但CD68、CD163、IL-6阳性细胞表达以及PGK1、HIF-1αmRNA表达差异无统计学意义(P>0.05)。与5FU组比较,H组小鼠的瘤体体积显著减小(P <0.01);肿瘤细胞侵袭、炎性细胞浸润、血管新生、细胞核分裂等病理表现明显改善;CD68、CD163、IL-6阳性细胞数均明显降低(P <0.001);PGK1 mRNA相对表达量显著降低(P <0.001);但HIF-1αmRNA的表达差异无统计学意义(P>0.05)。结论黄芪颗粒对H22荷瘤小鼠的化疗增效机制可能与其减少巨噬细胞浸润及M2型巨噬细胞极化,从而减少IL-6的分泌,抑制PGK1酶表达有关。Objective Based on energy metabolism,the mechanism of Astragalus granule on enhancing the efficacy of chemotherapy in H22 tumor-bearing mice was studied.Methods Eighteen Kunming mice were randomly divided into model group(M)and fluorouracil group(5 FU)and Astragalus granule group(H).H22 cell suspension(0.2 mL,1.5×107·mL-1)was subcutaneously injected into the right forearm axilla of mice to establish the liver cancer model.Mice in 5 FU and H groups were given 5 FU chemotherapy,once every day for 7 days.From the 8 th day,H group mice were given Astragalus granule once every day for 7 days.Positive cells expressing CD68,CD163,IL-6 in tumor tissues were visualized by immunohistochemical staining,and the numbers of positive cells were counted under light microscope for statistical analysis.Real-time quantitative PCR was used to detect the mRNA expression of PGK1 and HIF-1α.Results Compared with group M,the tumor volume of mice in the 5 FU group was significantly reduced(P<0.01).Neovascularization and mitotic images of invasive inflammatory cells infiltrating into surrounding tissues were significantly reduced.There were no statistically significant differences in the numbers of CD68,CD163,and IL-6 positive cells,and PGK1 and HIF-1αmRNA expression levels(P>0.05).Compared with the 5 FU group,the tumor volume in the H group was significantly reduced(P<0.01);tumor cells invation,inflammatory cells infiltration,angiogenesis and nuclear division,etc.were improved.The number of CD68,CD163,and IL-6 positive cells were significantly reduced(all P<0.001).The expression of PGK1 mRNA was decreased(P<0.001).However,there was no significant difference in HIF-1αmRNA expression(P>0.05).Conclusion The mechanism of synergistic effect of Astragalus granule on chemotherapy in H22 tumor-bearing mice may be related to its reduction of macrophage infiltration and the polarization of M2-type macrophages,thereby reducing the secretion of IL-6 and inhibiting the expression of the important glycolytic enzyme PGK1.

关 键 词：黄芪颗粒 肿瘤相关巨噬细胞 PGK1 IL-6 能量代谢 肝癌 化疗增效 

分 类 号：R285.5[医药卫生—中药学]