三七总皂苷溶液滴眼给药对大鼠非增殖性糖尿病视网膜病变的改善作用  

Ameliorative effect of Panax notoginseng saponins eye drops on non-proliferative diabetic retinopathy in rats

作　　者：孙鑫 王亚茹 程雪梅[1] 陈泓谕 陈明 赖树生 季莉莉[1] 尉小慧[1] 王长虹[1] SUN Xin;WANG Ya-ru;CHENG Xue-mei;CHEN Hong-yu;CHEN Ming;LAI Shu-sheng;JI Li-li;WEI Xiao-hui;WANG Chang-hong(The MOE Key Laboratory for Standardization of Chinese Medicines,Shanghai Key Laboratory for Compound Chinese Medicines,Institute of Chinese Materia Medica,Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Guangxi Key Laboratory of Comprehensive Utilization Technology of Pseudo-ginseng,Wuzhou 543000,China)

机构地区：[1]上海中医药大学中药研究所,中药标准化教育部重点实验室,上海市复方中药重点实验室,上海201203 [2]广西三七综合利用技术重点实验室,广西梧州543000

出　　处：《药学学报》2024年第5期1271-1279,共9页Acta Pharmaceutica Sinica

基　　金：上海市卫健委中医药传承创新发展三年行动计划建设项目[ZY(2021-2023)-0215];广西科技基地和人才专项(Guike AD20297068);上海中医药大学三七研究中心项目(SQZX202103)。

摘　　要：糖尿病视网膜病变(diabetic retinopathy,DR)是一种累及眼睛的糖尿病并发症,会导致患者视力低下甚至失明。本文旨在研究三七总皂苷(Panax notoginseng saponins,PNS)溶液滴眼给药对链脲佐菌素(streptozotocin,STZ)诱导的大鼠非增殖性糖尿病视网膜病变(non-proliferative diabetic retinopathy,NPDR)的改善作用及其机制。实验方案经上海中医药大学实验动物福利与伦理委员会审查,符合实验动物福利与伦理相关规范(伦理号:PZSHUTCM 211115004)。通过对大鼠腹腔单次注射STZ诱导糖尿病模型,注射两个月后,分别按照20、40和80 mg·kg-1的剂量双眼滴眼PNS溶液,每日2次,每次间隔6 h,连续1个月。通过对大鼠眼球进行苏木精-伊红染色和免疫组化分析,观察视网膜的形态结构变化和小胶质细胞的活化情况;采用伊文思蓝染料渗漏实验,观察血-视网膜屏障(blood-retina barrier,BRB)的破坏情况;利用胰酶消化视网膜并进行染色,观察视网膜中无细胞毛细血管的数量;通过尾静脉注射异硫氰酸荧光素偶联刀豆蛋白A凝集素,观察视网膜中黏附的中性粒细胞数量;采用酶联免疫实验检测血清中炎症因子的表达;采用Western blot实验检测视网膜组织中相关蛋白的表达以及核因子κB(nuclear factor kappa-B,NF-κB)的转录激活。结果发现,PNS溶液滴眼给药能够显著增加视网膜的厚度、减少无细胞毛细血管的数量以及上调紧密连接蛋白claudin-1和occludin的表达,从而缓解BRB的损伤。此外,PNS溶液滴眼给药也能够显著减少中性粒细胞的黏附和小胶质细胞的活化、降低血清中炎症因子的表达以及视网膜p65蛋白的核转移,有效抑制视网膜炎症的发生。以上结果表明,PNS溶液滴眼给药可以通过抑制NF-κB信号通路的激活,降低炎症的发生,从而发挥改善NPDR的作用。Diabetic retinopathy(DR) is a diabetic ocular complication that can lead to poor vision and blindness.This experiment aimed to investigate the ameliorative effect and its mechanism of Panax notoginseng saponins(PNS) eye drops on streptozotocin(STZ)-induced non-proliferative diabetic retinopathy(NPDR) in rats.All experiments were approved by the Animal Research Committee of Shanghai University of Traditional Chinese Medicine.Animal welfare and the animal experimental protocols were strictly consistent with related ethics regulations of Shanghai University of Traditional Chinese Medicine(No.PZSHUTCM 211115004).Diabetes mellitus was induced by a single intraperitoneal injection of STZ into rats.Two months later,PNS solution was dropped binocular twice per day at 6 h intervals at dose of 20,40,and 80 mg·kg-1 continuously for 1 month.The morphological structure and activation of microglia of the retina were observed by hematoxylin-eosin staining and immunohistochemical assay.The disruption of the blood-retina barrier(BRB) was conducted by the Evans blue dye leakage assay.The number of acellular capillaries in the retina was assessed by digesting and hematoxylineosin staining assay.The number of retinal leukocyte adhesion was observed by fluorescein isothiocyanate-coupled concanavalin A lectin labeling assay.The serum expression of inflammatory factors was measured using enzymelinked immunosorbent assay.Western blot experiments were used to detect the expression of relevant proteins in retinal tissue and transcriptional activation of nuclear factor kappa-B(NF-κB).The results revealed that PNS eye drops significantly increased the thickness of the retina,decreased the number of acellular capillaries,and up-regulated the expression of the tight junction proteins claudin-1 and occludin,thereby alleviating BRB damage.In addition,PNS eye drops were also able to significantly reduce leukocyte adhesion and microglia activation,the expression of inflammatory factors in serum,and the nuclear translocation of retinal p65 prote

关 键 词：三七总皂苷 滴眼给药 非增殖性糖尿病视网膜病变 血-视网膜屏障 炎症因子 核因子ΚB 

分 类 号：R966[医药卫生—药理学]