湖南中医药大学学报2025，Vol.45Issue(4)：598-607,10.DOI:10.3969/j.issn.1674-070X.2025.04.003
基于JAK-STAT/NF-κB通路探究黄精"生熟异治"治疗缺血性脑卒中大鼠的保护作用及分子机制研究
Protective effects and molecular mechanisms of"differential treatments of the raw and processed"Huangjing(Polygonati Rhizoma)on ischemic stroke rats based on JAK-STAT/NF-κB pathway
摘要
Abstract
Objective To investigate the protective effects and molecular mechanisms of raw and processed Huangjing(Polygonati Rhizoma)and their active components in treating ischemic stroke(IS)rats through network pharmacology and experimental studies.Methods A network pharmacology approach was used to construct a"key component-core target-pathway"network diagram for raw and processed Huangjing(Polygonati Rhizoma)in IS treatment.Fifty-six rats were randomly divided into sham-operated,model,nimodipine(20 mg/kg),low-dose raw Huangjing polysaccharide(0.1 g/kg),high-dose raw Huangjing polysaccharide(0.4 g/kg),low-dose processed Huangjing polysaccharide(0.1 g/kg),and high-dose processed Huangjing polysaccharide(0.4 g/kg)groups.Except for the sham-operated group,IS models were established via the suture-occluded method.After successful modeling,drugs were administered by gavage once daily for seven days.Longa neurological function was scored using the Zea Longa scale before administration and at 0.5,1,3,5,and 7 days after administration.After seven days,brain tissue was collected to calculate brain water content.Serum levels of interleukin(IL)-1β,IL-6,tumor necrosis factor-α(TNF-α),malondialdehyde(MDA),catalase(CAT),superoxide dismutase(SOD),ubiquitin C-terminal hydrolase L1(UCH-L1),neuron-specific enolase(NSE),and glial fibrillary acidic protein(GFAP)were measured by ELISA.Western blot was used to determine protein expression levels of phosphorylated Janus kinase 2/Janus kinase 2(p-JAK2/JAK2),phosphorylated signal transducer and activator of transcription 3/signal transducer and activator of transcription 3(p-STAT3/STAT3),phosphorylated nuclear factor-κB/nuclear factor-κB(p-NF-κB/NF-κB),B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),and caspase-3 in brain tissue.RT-qPCR was used to measure the relative mRNA expression levels of Bcl-2,Bax,and caspase-3.Results Network pharmacology identified protein kinase B α,cytochrome P450 family 2 subfamily D member 6,and STAT3 as common potential targets of raw and processed Huangjing(Polygonati Rhizoma)for IS treatment.Estrogen receptor α,dipeptidyl peptidase-4,and androgen receptor were unique targets for raw Huangjing(Polygonati Rhizoma),while caspase-3,metabotropic glutamate receptor 1,and solute carrier family 1 member 2 were potential targets for processed Huangjing(Polygonati Rhizoma).Oxidative stress response and inflammatory signaling transduction were common pathways for both.Compared with the sham-operated group,the model group showed increased brain water content and neurological function scores(P＜0.01);elevated serum IL-1β,IL-6,TNF-α,MDA,UCH-L1,NSE,and GFAP levels(P＜0.05);and significantly decreased CAT and SOD levels(P＜0.01).The protein expression levels of p-JAK2/JAK2,p-STAT3/STAT3,and p-NF-κB/NF-κB in brain tissue increased(P＜0.05),and the mRNA and protein expression levels of Bax and Caspase-3 elevated(P＜0.05),while the mRNA and protein expression level of Bcl-2 decreased(P＜0.05).Compared with the model group,both low-and high-dose raw and processed Huangjing polysaccharide groups showed reduced neurological function scores after five days of drug treatment(P＜0.01).Raw Huangjing polysaccharide groups had lower serum levels of IL-1β,IL-6,and TNF-α(P＜0.05,P＜0.01),while processed Huangjing polysaccharide groups showed reduced serum levels of MDA and UCH-L1(P＜0.05,P＜0.01).High-dose raw Huangjing polysaccharide group and processed Huangjing polysaccharide groups showed increases in serum SOD level(P＜0.05).High-dose processed Huangjing polysaccharide group showed decreases in p-JAK2/JAK2,p-STAT3/STAT3,and p-NF-κB/NF-κB protein expression levels(P＜0.05,P＜0.01),as well as mRNA expression levels of Bax and caspase-3 in brain tissue(P＜0.05,P＜0.01).High-dose raw and processed Huangjing polysaccharide groups exhibited elevated Bcl-2 mRNA expression level(P＜0.05).Conclusion Both raw and processed Huangjing polysaccharides can alleviate neural injury in IS rats,and the mechanism may involve reducing inflammatory cytokines,improving oxidative stress,lowering neural damage biomarkers expression,and modulating the JAK-STAT/NF-κB signaling pathway,thereby attenuating cerebral inflammatory reaction.关键词
生黄精/熟黄精/缺血性脑卒中/JAK-STAT/NF-κB通路/生熟异治Key words
raw Huangjing(Polygonati Rhizoma)/processed Huangjing(Polygonati Rhizoma)/ischemic stroke/JAK-STAT/NF-κB pathway/differential treatments of the raw and processed分类
中医学引用本文复制引用
刘红梅,周平兰,王竹鑫,彭敏,欧巧玲,邓蓝冰,周宜..基于JAK-STAT/NF-κB通路探究黄精"生熟异治"治疗缺血性脑卒中大鼠的保护作用及分子机制研究[J].湖南中医药大学学报,2025,45(4):598-607,10.基金项目
王竹鑫全国老药工传承工作室(国中医药人教函[2024]255号) （国中医药人教函[2024]255号）
第七批全国老中医药专家学术经验继承项目 （）
湖南省中医药管理局重点课题(A2023033) （A2023033）
湖南省自然科学基金科药联合项目(2021JJ80063) （2021JJ80063）
湖南省中医药管理局重点课题(C2022028). （C2022028）
