摘要
基于质谱成像法研究三七改善糖尿病视网膜病变(DR)及干预角膜、玻璃体和视网膜代谢物的作用,揭示三七改善DR作用机制。所有动物实验经北京中医药大学实验动物伦理委员会批准(批准号:BUCM-2023052204-2117)。采用链脲佐菌素(STZ)诱导糖尿病(DM)大鼠模型,检测各组大鼠的空腹血糖(FBG)和糖化血清蛋白(GSP)含量,应用免疫荧光染色法检测大鼠视网膜中闭合蛋白(occludin)、闭锁小带蛋白-1(ZO-1)表达水平;采用空气动力辅助解吸电喷雾离子化质谱成像(AFADESI-MSI)检测DM组和三七组大鼠眼球角膜、玻璃体、视网膜微区内源性代谢物,通过主成分分析(PCA)、正交偏最小二乘法判别分析(OPLS-DA)筛选DM组和三七组的差异代谢物,分析各微区中差异代谢物的原位空间信息,并通过京都基因与基因组百科全书(KEGG)数据库分析相关代谢通路。结果表明,与DM组比较,三七组糖尿病大鼠FBG和GSP均有下降趋势,视网膜中ZO-1、occludin表达增加(P<0.001);AFADESI-MSI分析结果显示,三七组角膜、玻璃体和视网膜微区共有34个差异代谢物,其中三七回调13种差异代谢物。在视网膜微区,三七显著回调溶血磷脂酰丝氨酸(18∶0)、磷脂酰乙醇胺(34∶2)和磷脂酰丝氨酸(40∶7/42∶7);代谢通路富集结果表明,三七主要调控甘油磷脂代谢、糖基磷脂酰肌醇合成、烟酸和烟酰胺代谢以及甘油酯代谢途径。综上,三七改善糖尿病大鼠血视网膜屏障(BRB),其作用机制可能与甘油磷脂代谢密切相关,本研究为三七改善DR的作用机制提供了科学依据,展现了质谱成像技术应用于药理机制研究的潜力。
Based on mass spectrometry imaging method,we investigated the effects of Panax notoginseng in improving diabetic retinopathy(DR)and interfering with corneal,vitreous and retinal metabolites,to reveal the mechanism of Panax notoginseng's action in improving DR.All animal experiments were approved by the Experimental Animal Ethics Committee of Beijing University of Chinese Medicine(Approval No.:BUCM2023052204-2117).Streptozotocin(STZ)-induced diabetes mellitus(DM)rat model was used,and fasting blood glucose(FBG)and glucosylated serum protein(GSP)levels were measured in each group of rats.Occludin and zonula occludens-1(ZO-1)were detected by immunofluorescence staining;air flow-assisted desorption electrospray ionization mass spectrometry imaging(AFADESI-MSI)was used to detect endogenous metabolites in the cornea,vitreous,and retinal microregions of the eyes of rats in the DM group and Panax notoginseng group.Endogenous metabolites were detected in the cornea,vitreous,and retinal microregions of the DM and Panax notoginseng groups,and the DM and Panax notoginseng groups were screened for different metabolites by principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA).Differential metabolites were screened in the DM and Panax notoginseng groups,the in situ spatial information of differential metabolites in each microregion was analyzed,and the related metabolic pathways were analyzed by the Kyoto encyclopedia of genes and genomes(KEGG)database.The results showed that compared with the DM group,diabetic rats in the Panax notoginseng group showed a decreasing trend in both FBG and GSP,and an increase in the expression of ZO-1 and occludin in the retina(P<0.001);AFADESI-MSI analysis showed that there were a total of 34 differential metabolites in the cornea,vitreous body,and retinal microregion in the Panax notoginseng group,of which Panax notoginseng called back 13 differential metabolites.In the retinal microregion,Panax notoginseng significantly regulated lysophosphatidylserine(18∶0),phosphatidylethanolamine(34∶2)and phosphatidylserine(40∶7/42∶7).The metabolic pathway enrichment results indicated that Panax notoginseng mainly regulated glycerophospholipid metabolism,glycosylphosphatidylinositol synthesis,niacin and nicotinamide metabolism as well as glycerol ester metabolic pathways.In conclusion,Panax notoginseng improves the blood-retinal barrier(BRB)in diabetic rats,and its mechanism of action may be closely related to glycerophospholipid metabolism.This study provides scientific evidence for the mechanism of action of Panax notoginseng in improving DR,and demonstrates the potential of mass spectrometry imaging technology applied to the study of pharmacological mechanisms.
作者
籍宇星
彭美中
宁尚秋
杨美美
刘卓容
张雨婷
郝改梅
韩静
JI Yu-xing;PENG Mei-zhong;NING Shang-qiu;YANG Mei-mei;LIU Zhuo-rong;ZHANG Yu-ting;HAO Gai-mei;HAN Jing(Beijing University of Chinese Medicine,Beijing 102488,China;Beijing Anzhen Hospital Affiliated to Capital Medical University,Beijing 100020,China;Institute of Basic Theory for Chinese Medicine,China Academy of Chinese Medical Sciences,Beijing 100700,China)
出处
《药学学报》
北大核心
2025年第5期1515-1524,共10页
Acta Pharmaceutica Sinica
基金
国家自然科学基金项目(82074238,81873165)。
关键词
三七
糖尿病视网膜病变
质谱成像
血视网膜屏障
甘油磷脂代谢
Panax notoginseng
diabetic retinopathy
mass spectrometry imaging
blood-retinal barrier
glycerophospholipid metabolism
