滋肾健脾化瘀片通过抑制AGEs/RAGE/NF-κB通路改善糖尿病大鼠的视网膜病变  

Zishen Jianpi Huayu Tablets Improve Retinopathy in Diabetic Rats by Inhibiting AGEs/RAGE/NF-κB Pathway

作　　者：李坤梦 许泽鹏 田妮[2] 董燕[3] 刘红[2] 俞晓艺[2] 余杨桂[2] 钟瑞英[2] 刘求红[2] 王小川[2] LI Kunmeng;XU Zepeng;TIAN Ni;DONG Yan;LIU Hong;YU Xiaoyi;YU Yanggui;ZHONG Ruiying;LIU Qiuhong;WANG Xiaochuan(The First Clinical Medical College of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;The First Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China;The Science and Technology Innovation Center of Guangzhou University of Chinese Medicine,Guangzhou 510405 Guangdong,China)

机构地区：[1]广州中医药大学第一临床医学院,广东广州510405 [2]广州中医药大学第一附属医院,广东广州510405 [3]广州中医药大学科技创新中心,广东广州510405

出　　处：《中药新药与临床药理》2020年第10期1147-1152,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广州中医药大学第一附属医院创新科研团队项目(2017TD04);广东省中医药局科研项目(20172048,20201286);广州市科技计划项目(201604020105);关国华全国名老中医药专家传承工作室(国中医药人教函[2018]134号)。

摘　　要：目的探讨滋肾健脾化瘀片改善糖尿病大鼠早期视网膜病变的作用机制。方法将28只雄性SD大鼠随机分为空白组、模型组及滋肾健脾化瘀片高、低剂量(2.4、0.6 g·kg^-1)组。采用腹腔注射2%链脲佐菌素(STZ,60 mg·kg^-1)建立糖尿病大鼠模型。灌胃给药(9.6 mL·kg^-1),每日1次,连续给药10周。采用HE染色法观察视网膜组织病理学变化,进行视网膜神经节细胞(RGC)计数;免疫组化法检测大鼠视网膜血管内皮生长因子(VEGF)、核因子κB(NF-κB)及糖基化终产物受体(RAGE)蛋白表达水平;ELISA法检测大鼠血清中白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、糖基化终产物(AGEs)水平。结果与空白组比较,模型组大鼠的视网膜神经节细胞层排列紊乱、内外核层水肿、空泡化改变并且视网膜的厚度变薄,RGC数量明显减少(P<0.05);视网膜VEGF、RAGE、NF-κB蛋白表达和血清AGEs、IL-1β、IL-6水平明显上调(P<0.05)。与模型组比较,滋肾健脾化瘀片高剂量组大鼠的视网膜层结构、排列以及水肿程度得到了一定改善,RGC数量明显增多(P<0.05),视网膜VEGF、RAGE、NF-κB蛋白表达和血清AGEs、IL-1β、IL-6水平明显下调(P<0.05);滋肾健脾化瘀片低剂量组大鼠的视网膜RGC数量亦明显增多(P<0.05),血清AGEs表达水平明显降低(P<0.05)。结论滋肾健脾化瘀片可改善糖尿病大鼠视网膜内、外核层的水肿情况,减少神经节细胞的丢失,其机制可能与抑制AGEs/RAGE/NF-κB通路来下调炎症因子的表达有关。Objective To explore the mechanism of Zishen Jianpi Huayu tablets in improving early retinopathy in diabetic rats.Methods 28 male SD rats were randomly divided into a blank group,a model group,a high-dose and low-dose groups(2.4,0.6 g·kg^-1 respectively).The blank group was injected intraperitoneally with the same amount of citrate buffer,and the remaining 3 groups were intraperitoneally injected with 2%streptozotocin(STZ)at a dose of 60 mg·kg^-1 to establish diabetic rat models.After successful modeling,the blank group and the model group were given the same amount of distilled water daily by gavage,and the other groups were given the intervention of gavage with Zishen Jianpi Huayu tablets.After 10 weeks,paraffin sections of the rats’retina were taken for HE staining to observe pathological changes and count the retinal ganglion cells(RGC);immunohistochemical were used to detect expression levels of retinal vascular endothelial growth factor(VEGF),nuclear factor kappa-B(NF-κB),and receptor for advanced glycation endproducts(RAGE).The levels of interleukin-1β(IL-1β),interleukin-6(IL-6),and advanced glycation end products(AGEs)in rats’serum were detected by ELISA.Results Compared with the blank group,in the model group,the arrangement of retinal ganglion cell layers was disordered,edema and vacuole changed in the inner and outer nuclear layers,and the thickness of the retina became thinner,and the number of RGCs was significantly reduced(P<0.05);serum expression levels of AGEs,IL-1β,IL-6 were significantly increased(P<0.05).Compared with the model group,the structure and arrangement of retinal layers and the degree of edema in the high-dose group decreased,and the number of RGCs increased significantly(P<0.05);the expression levels of retinal VEGF,RAGE,NF-κB and serum AGEs,IL-1β,IL-6 decreased significantly(P<0.05).Compared with the model group,in the low-dose group,the number of RGCs in the retina increased significantly(P<0.05),and the expression level of serum AGEs decreased significantly(P<0.05).Co

关 键 词：滋肾健脾化瘀片 糖尿病视网膜病变 糖基化终产物 糖基化终产物受体 核因子ΚB 大鼠 

分 类 号：R285.5[医药卫生—中药学]