膈下逐瘀汤对EMT气滞血瘀型大鼠NF-κB信号通路及其下游因子的影响  

Effect of Gexia Zhuyutang on NF-κB Signaling Pathway and Downstream Factors in EMT Rats with Qi Stagnation and Blood Stasis Syndrome

作　　者：王志平 崔轶凡 甘涛 WANG Zhiping;CUI Yifan;GAN Tao(Shanxi University of Chinese Medicine,Jinzhong 030619,China)

机构地区：[1]山西中医药大学,山西晋中030619

出　　处：《中国实验方剂学杂志》2022年第22期23-32,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：山西省科技厅应用基础研究计划面上自然基金项目(201901D111341)。

摘　　要：目的:探讨膈下逐瘀汤对子宫内膜异位症(EMT)气滞血瘀型大鼠的治疗作用及其机制研究。方法:从60只SPF级雌性未孕健康SD大鼠中随机抽取6只作为空白组,其余采用复合因素法构建气滞血瘀型EMT大鼠模型。从造模成功的36只大鼠中随机抽取30只分为模型组、甲羟孕酮组(0.416 mg·kg^(-1))及膈下逐瘀汤低、中、高剂量组(4.063、8.125、16.25 g·kg^(-1)),每组6只,每天灌胃1次,连续4周。通过大鼠病理学变化、异位灶体积反映大鼠造模情况及膈下逐瘀汤对EMT大鼠的影响;取各组内膜组织进行苏木素-伊红(HE)染色,观察组织病理变化;酶联免疫吸附测定法(ELISA)检测各组血清上清液中白细胞介素-10(IL-10)、白细胞介素-6(IL-6)含量;免疫组化(IHC)检测各组大鼠组织中核转录因子-κB(NF-κB)、基质金属蛋白酶-9(MMP-9)、血管内皮生长因子(VEGF)、B细胞淋巴瘤-2(Bcl-2)相关X蛋白(Bax)蛋白表达情况;实时荧光定量聚合酶链式反应(Real-time PCR)检测NF-κB、MMP-9、VEGF、Bax mRNA表达。结果:治疗后,膈下逐瘀汤低、中、高剂量组和甲羟孕酮组异位灶体积明显缩小;镜下病理观察可见假手术组与空白组大鼠子宫内膜结构完整,腺体及间质细胞生长良好,腺上皮细胞排列紧密整齐,胞浆丰富呈高柱状;间质细胞分布均匀,血管丰富,呈梭形。与空白组、假手术组比较,模型组以柱状上皮和立方上皮为主,异位内膜上皮细胞呈短柱状,部分为假层状,细胞形态不完整,间质细胞和腺体的数量减少,部分腺体呈圆形,蜕膜发育不良。与空白组比较,模型组血清中IL-10含量明显降低(P<0.05),IL-6含量明显升高(P<0.05),模型组在位、异位内膜组织中NF-κB、VEGF、MMP-9蛋白表达明显升高(P<0.05),Bax蛋白表达明显降低;与模型组比较,膈下逐瘀汤中、高剂量组血清中IL-10含量明显升高(P<0.05),IL-6含量明显降低(P<0.05),膈下逐瘀汤各剂量组大鼠在位、异�Objective:To investigate the therapeutic effect of Gexia Zhuyutang on rats with endometriosis(EMT)with Qi stagnation and blood stasis syndrome and its mechanism.Method:Six female unpregnant healthy SD rats of SPF grade were randomly selected as a blank group.The rest were treated with the compound factor method to induce EMT with Qi stagnation and blood stasis syndrome in rats.After successful modeling,30 rats from 36 rats were randomly divided into a model group,a medroxyprogesterone(0.416 mg·kg^(-1))group,and low,medium,and high-dose Gexia Zhuyutang groups(4.063,8.125,16.25 g·kg^(-1)),with 6 rats in each group.Each group was given corresponding drugs,once a day for 4 consecutive weeks.The effect of Gexia Zhuyutang on rats with EMT was observed by the pathological changes and the volume of ectopic foci.The endometrial tissues of each group were taken for hematoxylin-eosin(HE)staining to observe histopathological changes.Enzyme-linked immunosorbent assay(ELISA)was used to determine the content of interleukin(IL)-10 and IL-6 in the serum supernatant of each group.Immunohistochemistry(IHC)was used to determine the protein expressions of nuclear factor kappa B(NF-κB),matrix metalloproteinase-9(MMP-9),vascular endothelial growth factor(VEGF),and Bcl-2 related X protein(Bax)in the tissues of rats in each group.Real-time fluorescence quantitative reverse transcription polymerase chain reaction(Real-time PCR)was used to determine the mRNA expressions of NF-κB,MMP-9,VEGF,and Bax.Result:After treatment,the volume of ectopic foci in the low,medium,and high-dose Gexia Zhuyutang groups and the medroxyprogesterone group was significantly reduced.The pathological observation under the microscope showed that the endometrial structure of rats in the sham-operated group and the blank group was intact,glands and mesenchymal cells grew well,glandular epithelial cells were arranged tightly and neatly,the cytoplasm was abundant and highly columnar,the interstitial cells were evenly distributed,and the blood vessels were abundant

关 键 词：膈下逐瘀汤 子宫内膜异位症 核转录因子-κB(NF-κB)信号通路 炎症反应 免疫抑制 微血管新生 

分 类 号：R2-0[医药卫生—中医学] R33R289R271.1