摘要
目的探索三黄四物汤(SST)不同溶剂提取物的体外抗炎作用及其相关机制,分析其主要成分与抗炎作用的关系,并探索其体内抗炎作用。方法通过制备三黄四物汤水提物(SSTW)及乙醇提取物(95%乙醇提取物SSTE、50%乙醇提取物SSTWE);采用高效液相色谱法(HPLC)分析其主要成分;通过LPS诱导RAW264.7细胞建立炎症模型,并采用苯酚胶浆法建立大鼠慢性盆腔炎(chronic pelvic inflammatory disease,CPID)模型;使用MTT法检测细胞存活率,Griess法检测NO水平,ELISA法检测PGE2、IL-6、TNF-α水平,Western blot法检测COX-2、iNOS及MAPKs通路、NF-κB通路相关蛋白表达水平,并通过病理切片观察大鼠子宫病理改变。结果SSTWE 0.78-25μg/mL组与正常组相比,细胞存活率无显著差异(P>0.05)。SSTWE 12.5μg/mL抗炎作用弱于SSTE而类似于SSTW,SSTWE 25μg/mL抗炎作用强于SSTW而类似于SSTE。SSTW、SSTE和SSTWE均明显降低LPS诱导的RAW264.7细胞iNOS和COX-2蛋白表达(P<0.05);SSTWE明显降低LPS诱导的RAW264.7细胞ERK磷酸化蛋白表达、IκB-α磷酸化蛋白表达及细胞核中p65蛋白表达,同时增加IκB-α总蛋白表达及细胞质中p65蛋白的表达水平(P<0.05)。SST明显抑制CPID大鼠血清TNF-α和IL-6含量(P<0.05),减轻盆腔局部组织粘连,改善子宫病理形态。结论三黄四物汤有显著体外抗炎作用,其机制均与炎症通路MAPKs和NF-κB有关;其不同提取物抗炎作用及细胞毒性不同,可能与当归成分有关;SST对大鼠CPID有显著抗炎作用,可作为慢性盆腔炎的候选药物。
Objective To explore the in vitro anti-inflammatory effects and related mechanisms of different solvent extracts of Sanhuang-Siwu-Tang(SST),analyze the relationship between its main components and anti-inflammatory effects,and investigate its in vivo anti-inflammatory effects.Methods SST water extract(SSTW),as well as 95%ethanol extract(SSTE)and 50%ethanol extract(SSTWE)were prepared.High-Performance Liquid Chromatography(HPLC)was used to analyze the main components.An inflammatory model was established by inducing RAW264.7 cells with LPS,and a rat model of chronic pelvic inflammatory disease(CPID)was established by phenol mucilage method.Cell viability was determined by MTT;NO level was measured by Griess;PGE2,IL-6,and TNF-αlevels were examined by ELISA;COX-2,iNOS,MAPKs,and NF-κB protein expressions were measured by Western blot;pathological changes in the rat uterus were observed through pathological sections.Results There was no significant difference in cell viability between the SSTWE groups at various concentrations from 0.78 to 25μg/mL and normal group(P>0.05).The anti-inflammatory effects of SSTWE at 12.5μg/mL were weaker than those of SSTE but similar to those of SSTW,while the anti-inflam-matory effects of SSTWE at 25μg/mL were stronger than those of SSTW but similar to those of SSTE.SSTW,SSTE,and SSTWE all significantly reduced iNOS and COX-2 protein expressions in LPS-induced RAW264.7 cells(P<0.05).SSTWE significantly reduced the protein expressions of p-ERK,p-IκB-α,and nuclear p65 in LPS-induced RAW264.7 cells,while increasing IκB-αtotal protein expression and cytosolic p65 protein expression(P<0.05).SST significantly lowered serum TNF-αand IL-6 levels in CPID rats(P<0.05),reduced local pelvic tissue adhesion,and improved pathological morphology of the uterus.Conclusion SST has significant in vitro anti-inflammatory effects,and its mechanisms are related to the inflammatory pathways of MAPKs and NF-κB.Its different extracts have varying anti-inflammatory effects and cytotoxicity,possibly r
作者
李静
余涛
齐耀群
张小容
戢佳慧
李汉超
LI Jing;YU Tao;QI Yaoqun;ZHANG Xiaorong;JI Jiahui;LI Hanchao(Affiliated Hospital of Jiujiang University,Jiujiang,Jiangxi 332000,China)
出处
《湖南中医药大学学报》
CAS
2024年第11期2024-2031,共8页
Journal of Hunan University of Chinese Medicine
基金
江西省教育厅科学技术青年项目(GJJ211841)
江西省卫生健康委员会科技计划(SKJP220219734)
江西省中医药管理局科技计划(2022B1032)。
关键词
三黄四物汤
炎症
慢性盆腔炎
信号通路
提取物
Sanhuang-Siwu-Tang
inflammation
chronic pelvic inflammatory disease
signaling pathway
extract
