基于斑马鱼模型的三物黄芩汤抗炎活性及机制研究  

Anti-inflammatory effects and mechanisms of Sanwu Huangqin Decoction based upon zebrafish models

作　　者：舒余琪 杨小瑜 彭颖慧 汤庆发 曹惠慧 谢涛 邵萌 SHU Yuqi;YANG Xiaoyu;PENG Yinghui;TANG Qingfa;CAO Huihui;XIE Tao;SHAO Meng(Guangdong Provincial Key Laboratory of Chinese Medicine Pharmaceutics,School of Traditional Chinese Medicine,Southern Medical University,Guangdong Guangzhou 510515,China;School of Pharmacy,Jinan University,Guangdong Guangzhou 510632,China;Department of Radiation Oncology,Zhujiang Hospital,Southern Medical University,Guangdong Guangzhou 510280,China)

机构地区：[1]南方医科大学中医药学院,广东省中药制剂重点实验室,广东广州510515 [2]暨南大学药学院,广东广州510632 [3]南方医科大学珠江医院肿瘤放疗科,广东广州510280

出　　处：《中国医院药学杂志》2024年第21期2433-2439,共7页Chinese Journal of Hospital Pharmacy

基　　金：国家自然科学基金项目(编号:82274169);广东省自然科学基金项目(编号:2023A1515011111);广东省中医药管理局项目(编号:20221261,20241215)。

摘　　要：目的:基于斑马鱼炎症模型探讨三物黄芩汤对急性炎症的抑制作用及机制。方法:采用硫酸铜浸泡法、尾部横切法和脂多糖(LPS)卵黄囊显微注射法构建3种斑马鱼急性炎症模型,分为对照组,模型组,阳性药组(10μg·mL^(-1)地塞米松),三物黄芩汤低、中、高剂量(100、200、300μg·mL^(-1))组,荧光观察各组斑马鱼损伤部位中性粒细胞的聚集。在LPS诱导炎症模型中,苏木精-伊红染色(HE染色)观察斑马鱼体内炎症细胞的数量变化;实时荧光定量PCR(RT-qPCR)技术测定炎症相关基因的转录水平;乳酸试剂盒检测斑马鱼体内的乳酸浓度。结果:与模型组相比,三物黄芩汤各给药组斑马鱼炎症部位中性粒细胞聚集数量均显著降低。LPS诱导的斑马鱼在给予三物黄芩汤后存活率显著提升,肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)和核因子-κB(NF-κB)的mRNA水平显著下调,乳酸脱氢酶A(LDHA)和IL-10的mRNA水平以及乳酸浓度显著上调。结论:三物黄芩汤能够调控LDHA介导的NF-κB信号通路,有效抑制中性粒细胞聚集,进而缓解急性炎症进程。OBJECTIVE To explore the inhibitory effects and mechanisms of Sanwu Huangqin Decoction(SWHD) on acute inflammation based upon zebrafish inflammatory models.METHODS CuSO_(4) immersion,tail transection and lipopolysaccharide(LPS) yolk injection zebrafish models were set up for evaluating the acute anti-inflammatory effects of SWHD.Zebrafish larvae were divided into four groups of control,model,positive(10 μg·mL^(-1) dexamethasone) and SWHD treatment(100/200/300 μg·mL^(-1)).Aggregation of neutrophils in injured area of zebrafish was observed by fluorescence.In LPS-induced inflammation model,hematoxylin-eosin(HE) stain was utilized for observing the changes in the number of inflammatory cells in zebrafish.Quantitative reverse-transcription polymerase chain reaction(qRT-PCR) was employed for measuring the transcript mRNA levels of inflammation-related genes and lactate kit for detecting lactate concentration in zebrafish.RESULTS As compared with model group,SWHD dosing blunted abnormal neutrophil recruitment to inflamed sites.The survival rate of LPS-induced zebrafish was significantly elevated after dosing of SWHD.Significantly elevated lactate level,up-regulated mRNA expressions of lactate dehydrogenase A(LDHA) and interleukin-10(IL-10) and down-regulated mRNA expressions of NF-κB,IL-6 and TNF-αwere also observed.CONCLUSION SWHD may effectively suppress neutrophil aggregation to alleviate acute inflammatory process through regulating the LDHA-mediated NF-κB signaling pathway.

关 键 词：三物黄芩汤 斑马鱼 炎症 NF-ΚB 乳酸脱氢酶 

分 类 号：R285[医药卫生—中药学] R966[医药卫生—中医学]