Hou Xiangmei, Zhang Ziying, Ma Yuehong, Jin Rong, Yi Bing, Yang Dongdong, Ma Lijie
Inner Mongolia Key Laboratory of Molecular Biology, Inner Mongolia Medical University, Hohhot, China.
Front Pharmacol. 2022 Sep 2;13:966759. doi: 10.3389/fphar.2022.966759. eCollection 2022.
To investigate how Hydroxysafflor yellow A (HSYA) effects acute liver injury (ALI) and what transcriptional regulatory mechanisms it may employ. Rats were randomly divided into five groups ( = 10): Control, Model, HSYA-L, HSYA-M, and HSYA-H. In the control and model groups, rats were intraperitoneally injected with equivalent normal saline, while in the HSYA groups, they were also injected with different amounts of HSYA (10, 20, and 40 mg/kg/day) once daily for eight consecutive days. One hour following the last injection, the control group was injected into the abdominal cavity with 0.1 ml/100 g of peanut oil, and the other four groups got the same amount of a peanut oil solution containing 50% CCl. Liver indexes were detected in rats after dissection, and hematoxylin and eosin (HE) dyeing was utilized to determine HSYA's impact on the liver of model rats. In addition, with RNA-Sequencing (RNA-Seq) technology and quantitative real-time PCR (qRT-PCR), differentially expressed genes (DEGs) were discovered and validated. Furthermore, we detected the contents of anti-superoxide anion (anti-O ) and hydrogen peroxide (HO), and verified three inflammatory genes (Icam1, Bcl2a1, and Ptgs2) in the NF-kB pathway by qRT-PCR. Relative to the control and HSYA groups, in the model group, we found 1111 DEGs that were up-/down-regulated, six of these genes were verified by qRT-PCR, including Tymp, Fabp7, Serpina3c, Gpnmb, Il1r1, and Creld2, indicated that these genes were obviously involved in the regulation of HSYA in ALI model. Membrane rafts, membrane microdomains, inflammatory response, regulation of cytokine production, monooxygenase activity, and iron ion binding were significantly enriched in GO analysis. KEGG analysis revealed that DEGs were primarily enriched for PPAR, retinol metabolism, NF-kB signaling pathways, etc. Last but not least, compared with the control group, the anti-O content was substantially decreased, the HO content and inflammatory genes (Icam1, Bcl2a1, and Ptgs2) levels were considerably elevated in the model group. Compared with the model group, the anti-O content was substantially increased, the HO content and inflammatory genes (Icam1, Bcl2a1, and Ptgs2) levels were substantially decreased in the HSYA group ( < 0.05). HSYA could improve liver function, inhibit oxidative stress and inflammation, and improve the degree of liver tissue damage. The RNA-Seq results further verified that HSYA has the typical characteristics of numerous targets and multiple pathway. Protecting the liver from damage by regulating the expression of Tymp, Fabp7, Serpina3c, Gpnmb, Il1r1, Creld2, and the PPAR, retinol metabolism, NF-kappa B signaling pathways.
为研究羟基红花黄色素A（HSYA）对急性肝损伤（ALI）的影响及其可能采用的转录调控机制。将大鼠随机分为五组（每组n = 10）：对照组、模型组、HSYA-L组、HSYA-M组和HSYA-H组。对照组和模型组大鼠腹腔注射等量生理盐水，而HSYA组大鼠连续八天每天腹腔注射不同剂量的HSYA（10、20和40 mg/kg/天）。末次注射后1小时，对照组腹腔注射0.1 ml/100 g花生油，其他四组注射等量含50% CCl₄的花生油溶液。解剖大鼠后检测肝脏指标，采用苏木精-伊红（HE）染色法观察HSYA对模型大鼠肝脏的影响。此外，利用RNA测序（RNA-Seq）技术和定量实时聚合酶链反应（qRT-PCR）发现并验证差异表达基因（DEGs）。进一步检测抗超氧阴离子（抗-O₂⁻）和过氧化氢（H₂O₂）含量，并通过qRT-PCR验证核因子κB（NF-κB）通路中的三个炎症基因（Icam1、Bcl2a1和Ptgs2）水平。与对照组和HSYA组相比，模型组发现1111个上调/下调的DEGs，其中6个基因经qRT-PCR验证，包括Tymp、Fabp7、Serpina3c、Gpnmb、Il1r1和Creld2，表明这些基因明显参与HSYA对ALI模型的调控。基因本体（GO）分析显示，膜筏、膜微区、炎症反应、细胞因子产生的调控、单加氧酶活性和铁离子结合等功能显著富集。京都基因与基因组百科全书（KEGG）分析表明，DEGs主要富集于过氧化物酶体增殖物激活受体（PPAR）、视黄醇代谢、NF-κB信号通路等。最后，与对照组相比，模型组抗-O₂⁻含量显著降低，H₂O₂含量和炎症基因（Icam1、Bcl2a1和Ptgs2）水平显著升高。与模型组相比，HSYA组抗-O₂⁻含量显著增加，H₂O₂含量和炎症基因（Icaml、Bcl2a1和Ptgs2）水平显著降低（P < 0.05）。HSYA可改善肝功能，抑制氧化应激和炎症反应，减轻肝组织损伤程度。RNA-Seq结果进一步证实HSYA具有多靶点、多通路的典型特征，通过调节Tymp、Fabp7、Serpina3c、Gpnmb、Il1r1、Creld2的表达以及PPAR、视黄醇代谢、NF-κB信号通路发挥肝脏保护作用。
