加味芍药甘草汤对他莫昔芬诱导的子宫腺肌病小鼠血管生成相关因子的影响  

Effect of Jiawei Shaoyao Gancao Decoction on Angiogenesis-Related Factors in Tamoxifen-Induced Mice with Adenomyosis

作　　者：温小慧 刘学红 黄诗雅 关永格 李坤寅 WEN Xiaohui;LIU Xuehong;HUANG Shiya;GUAN Yongge;LI Kunyin(The Third Clinical Medical School of Guangzhou University of Chinese Medicine,Guangzhou 510006 Guangdong,China;Lingnan Medical Research Center,Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;The Third Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)

机构地区：[1]广州中医药大学第三临床医学院,广东广州510006 [2]广州中医药大学岭南医学研究中心,广东广州510405 [3]广州中医药大学第三附属医院,广东广州510100

出　　处：《中药新药与临床药理》2025年第4期515-521,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广东省普通高校重点领域专项项目(2021ZDZX2015,2023ZDZX2011);广州中医药大学“双一流”与高水平大学学科协同创新团队项目;广州中医药大学中医学学科中医基础理论研究“揭榜挂帅”项目。

摘　　要：目的探讨加味芍药甘草汤(JSGD)对他莫昔芬诱导的子宫腺肌病(AM)小鼠血管内皮生长因子(VEGF)、缺氧诱导因子1α(HIF-1α)及微血管密度(MVD)的影响。方法将36只雌性ICR新生小鼠作为研究对象,随机选取6只作为正常组,其余30只作为造模组。在出生后第2~5天,造模组小鼠给予2.7μmol·kg^(-1)他莫西芬+5μL·g^(-1)花生油、卵磷脂、炼乳混合液(体积比=2∶0.2∶3)滴喂以建立AM小鼠模型。第16周模型复制成功后,将24只小鼠随机分为模型组、JSGD高剂量组(5.972 g·kg^(-1)·d^(-1))、JSGD低剂量组(2.986 g·kg^(-1)·d^(-1))及地诺孕素组(0.362 mg·kg^(-1)·d^(-1)),每组6只,灌胃给药,每天1次,连续3周。采用HE染色法观察小鼠子宫组织病理变化,并按照AM标准分级进行评分;比色法检测血清ALT、AST水平,ELISA法检测血清VEGF水平;RT-PCR法检测子宫组织中VEGF、HIF-1αmRNA表达水平;免疫荧光法检测子宫组织中VEGF、CD31的表达水平。结果各组小鼠的血清ALT、AST水平无明显差异(P>0.05),表明他莫昔芬造模方法及JSGD对AM小鼠的ALT、AST水平无影响。与正常组比较,模型组小鼠子宫内膜侵入程度评分显著升高(P<0.01),提示AM模型小鼠复制成功;血清VEGF水平显著升高(P<0.01);子宫组织VEGF、HIF-1αmRNA表达均显著上调(P<0.05,P<0.01),VEGF蛋白及MVD表达水平均显著升高(P<0.01)。与模型组比较,JSGD高、低剂量组及地诺孕素组小鼠子宫内膜侵入程度评分均明显降低(P<0.05);JSGD低剂量组小鼠的血清VEGF水平明显降低(P<0.05);JSGD高、低剂量组及地诺孕素组小鼠子宫组织VEGF mRNA表达均显著下调(P<0.01),VEGF蛋白及MVD表达水平均显著降低(P<0.01)。结论JSGD可以抑制他莫昔芬诱导的AM小鼠子宫组织的VEGF、HIF-1α及CD31表达,降低MVD水平,通过调控血管生成发挥治疗AM的作用。Objective To investigate the effects of Jiawei Shaoyao Gancao Decoction(JSGD)on vascular endothelial growth factor(VEGF),hypoxia-inducible factor-1α(HIF-1α)and microvessel density(MVD)in Tamoxifeninduced adenomyosis(AM)mice.Methods A total of 36 female ICR neonatal mice were selected as the research objects.There were 6 mice randomly selected as the normal group,and the remaining 30 mice were used as the model group.After 2 days and 5 days of birth,the mice in the model group were given 2.7μmol·kg^(-1)Tamoxifen+5μL·g^(-1)peanut oil,lecithin and condensed milk mixture(volume ratio=2∶0.2∶3)to establish AM mice model.After the successful replication of the model in the week 16,24 mice were randomly divided into model group,JSGD high-dose group(5.972 g·kg^(-1)·d^(-1)),JSGD low-dose group(2.986 g·kg^(-1)·d^(-1))and Dienogest group(0.362 mg·kg^(-1)·d^(-1)),with 6 mice in each group.They were given intragastric administration once a day for 3 consecutive weeks.The pathological changes of uterine tissue in mice were observed by HE staining and scored according to AM standard grading.Serum ALT and AST levels were detected by colorimetry,and serum VEGF levels were detected by ELISA.The mRNA expression levels of VEGF and HIF-1αin uterine tissue were detected by RT-PCR.The expression levels of VEGF and CD31 in uterine tissues were detected by immunofluorescence.Results There was no significant difference in serum ALT and AST levels in each group(P>0.05),indicating that Tamoxifen modeling method and JSGD had no effect on ALT and AST levels in AM mice.Compared with the normal group,the score of endometrial invasion in the model group was significantly increased(P<0.01),suggesting that the AM model mice were successfully replicated.Serum VEGF level was significantly increased(P<0.01).The mRNA expressions of VEGF and HIF-1αin uterine tissue were significantly up-regulated(P<0.05,P<0.01).The expression levels of VEGF and MVD in uterine tissue were significantly increased(P<0.01).Compared with the model group,the

关 键 词：加味芍药甘草汤 子宫腺肌病 血管生成 微血管密度 血管内皮生长因子 缺氧诱导因子1Α 小鼠 

分 类 号：R285.5[医药卫生—中药学]