杞菊地黄丸调控Raf/MEK/ERK通路改善糖尿病视网膜病变小鼠炎症反应实验研究  

Experimental Study of Qiju Dihuang Pills(杞菊地黄丸)Improving Inflammatory Response in Mice with Diabetic Retinopathy by Regulating Raf/MEK/ERK Pathway

作　　者：赵春林 马广成 陶思翰 陈卓琳 王明月 方祎鸣 施炜 ZHAO Chunlin;MA Guangcheng;TAO Sihan;CHEN Zhuolin;WANG Mingyue;FANG Yiming;SHI Wei(First College of Clinical Medicine,Nanjing University of Chinese Medicine,Nanjing 210023,Jiangsu,China;Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210004,Jiangsu,China)

机构地区：[1]南京中医药大学第一临床医学院,江苏南京210023 [2]南京中医药大学附属医院,江苏南京210004

出　　处：《中华中医药学刊》2025年第1期152-156,I0029-I0031,共8页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81904256);江苏省中医药科技发展计划项目(YB201953);溧阳市科技项目(LC2019001)。

摘　　要：目的基于纤维肉瘤蛋白(rapid accelerated fibrosarcoma,Raf)/丝裂原活化的细胞外信号调节激酶(mitogen-activated extracellular signal-regulated kinase,MEK)/细胞外信号调节激酶(extracellular signal regulated kinase,ERK)信号通路探讨杞菊地黄丸改善糖尿病视网膜病变炎症反应的价值。方法将50只雄性C57BL/6J小鼠随机分为正常组,模型组,杞菊地黄丸低剂量组,杞菊地黄丸中剂量组以及杞菊地黄丸高剂量组。通过腹腔注射链脲佐菌素(streptozotocin,STZ)建立糖尿病模型。造模12周后,正常组、模型组每天予生理盐水灌胃1次,杞菊地黄丸各剂量组每天予相应杞菊地黄丸药液(11,22以及44 g·kg^(-1))灌胃1次,连续8周。每月测定血糖、饮水量、排尿量、进食量以及体质量变化;影像学检测[眼底血管荧光造影(Fundus angiofluorescence,FFA)、光学相干断层扫描血管成像(Optical coherence tomography angiography,OCTA)以及光学相干断层扫描(Optical coherence tomography,OCT)]评估视网膜病变情况;苏木精-伊红(Hematoxylin-eosin,HE)染色评估视网膜组织病理学变化;免疫荧光(immunofluorescence,IF)检测视网膜组织白介素-6(interleukin-6,IL-6),白介素-17(interleukin-17,IL-17)以及肿瘤坏死因子α(tumor necrosis factorα,TNF-α)表达;酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测外周血中IL-6、IL-17以及TNF-α水平。蛋白免疫印迹法(western blot,WB)检测视网膜组织中Raf、p-Raf、MEK、p-MEK、ERK以及p-ERK表达。结果与正常组相比,模型组小鼠随机血糖高,多饮、多尿、多食以及体质量下降症状明显;视网膜血管密度低,管径增粗,走行迂曲;视网膜各层结构疏松,总体厚度变薄明显;IL-6、IL-17、TNF-α、p-Raf、p-MEK以及p-ERK表达明显增多(P<0.01)。与模型组相比,杞菊地黄丸组小鼠随机血糖高,多饮、多尿、多食以及体质量下降症状明显改善;血管密度低,管径增粗,走行迂曲程度明显减轻;视�Objective To explore the effects of Qiju Dihuang Pills(杞菊地黄丸)on inflammatory responses in mice with diabetic retinopathy based on the rapid accelerated fibrosarcoma(Raf)/mitogen-activated extracellular signal-regulated kinase(MEKERK)signaling pathway.Methods Fifty male C57BL/6J mice were randomly divided into the normal group,the model group,the Qiju Dihuang Pills low-dose,middle-dose and high-dose groups.The model was established by intraperitoneal injection of streptozotocin(STZ).After 12 weeks of modeling,the normal group and the model group were given saline by gavage once a day,and the Qiju Dihuang Pills Groups were given the corresponding dose of Qiju Dihuang Pills medicinal solution(11 g·kg^(-1),22 g·kg^(-1)and 44 g·kg^(-1))once a day for 8 consecutive weeks.The changes in blood glucose,water intake,urine output,food intake and body mass were measured monthly.The imaging tests[fundus angiofluorescence(FFA),optical coherence tomography angiography(OCTA)and optical coherence tomography(OCT)]were performed to assess the retinopathy.HE staining was used to assess retinal histopathological changes and immunofluorescence(IF)to detect the expressions of interleukin-6(IL-6),interleukin-17(IL-17)and tumor necrosis factorα(TNF-α).ELISA was used to detect the levels of IL-6,IL-17 and TNF-αin peripheral blood.Western Blot was used to detect the expressions of Raf,p-Raf,MEK,p-MEK,ERK and p-ERK in retinal tissues.Results Compared with those of the normal group,the mice in the model group had high blood sugar at random,obvious symptoms of polydipsia,polyuria,polyphagia and weight loss;the thickness was significantly thinner.The expressions of IL-6,IL-17,TNF-α,p-Raf,p-MEK and p-ERK were significantly increased(P<0.01).Compared with those in the model group,the mice in the Qiju Dihuang Pills groups had high random blood sugar,and the symptoms of polydipsia,polyuria,polyphagia and weight loss were significantly improved.The blood vessel density was low,the diameter of the vessel was thickened,and the degree

关 键 词：糖尿病视网膜病变 杞菊地黄丸 炎症 Raf/MEK/ERK通路 

分 类 号：R289.5[医药卫生—方剂学]