Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China; Chinese Institute for Brain Research, Beijing, 102206, China.
Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China.
Phytomedicine. 2021 Nov;92:153613. doi: 10.1016/j.phymed.2021.153613. Epub 2021 May 28.
Transcription factors (TFs) play a critical role in the cerebral ischemia/reperfusion injury (IRI). Panax notoginseng saponins (PNS) are extensively used in the treatment of acute cerebral ischemia in China, but the mechanism of their effects, especially at the TF level, remains unclear. In this study, a combination of transcriptomics, proteomics and network pharmacology analysis was used to identify the key TFs involved in the protection of PNS against middle cerebral artery occlusion (MCAO)-induced IRI.
Sprague-Dawley rats which were subjected to 1.5 hours of MCAO-induced occlusionand then followed by reperfusion, were treated with PNS at a concentration of 36 mg/kg or 72 mg/kg daily for 7 days. PNS significantly decreased neurological deficient scores and infarction rate; prevented cerebral tissue damage; and reduced CASP3 activity, levels of TNF, IL1B and CCL2 after IRI. Through a combination of transcriptomics and proteomics, 9 critical TFs were identified, including Excision repair cross-complementing group 2 (ERCC2), Nuclear receptor subfamily 4 group A member 3 (NR4A3) and 7 other TFs. The targets of ERCC2 and NR4A3, such as Ubxn11, Ush2a, Numr2, Oxt, Ubxn11, Scrt2, Ttc34 and Lrrc23, were verified by using real-time PCR analysis. RNA-seq analyses indicated that PNS regulated nerve system development and inflammation, and the majority of the identified TFs were also involved in these processes. By using network pharmacology analysis, 73 chemical components in PNS were predicted to affect ERCC2, NR4A3 and 3 other identified TFs.
ERCC2, NR4A3 and 7 other TFs were of importance in the protection of PNS against IRI. This study promoted the understanding of protective mechanism of PNS against cerebral IRI and facilitated the identification of possible targets of PNS.
转录因子（TFs）在脑缺血/再灌注损伤（IRI）中发挥关键作用。三七总皂苷（PNS）在中国广泛用于治疗急性脑缺血，但它们的作用机制，特别是在 TF 水平上，仍不清楚。在这项研究中，我们采用转录组学、蛋白质组学和网络药理学分析相结合的方法，鉴定了 PNS 对大脑中动脉闭塞（MCAO）诱导的 IRI 保护作用所涉及的关键 TF。
将 Sprague-Dawley 大鼠进行 1.5 小时 MCAO 诱导的闭塞，然后再进行再灌注，然后用 36 或 72mg/kg 的 PNS 浓度进行治疗，每天 1 次，共 7 天。PNS 可显著降低神经功能缺损评分和梗死率；防止脑组织损伤；降低 IRI 后 CASP3 活性、TNF、IL1B 和 CCL2 的水平。通过转录组学和蛋白质组学的结合，鉴定出 9 个关键 TF，包括切除修复交叉互补组 2（ERCC2）、核受体亚家族 4 组 A 成员 3（NR4A3）和其他 7 个 TF。通过实时 PCR 分析验证了 ERCC2 和 NR4A3 的靶点，如 Ubxn11、Ush2a、Numr2、Oxt、Ubxn11、Scrt2、Ttc34 和 Lrrc23。RNA-seq 分析表明，PNS 调节神经系统发育和炎症，而大多数鉴定出的 TF 也参与了这些过程。通过网络药理学分析，预测 PNS 中的 73 种化学成分可能影响 ERCC2、NR4A3 和其他 3 个鉴定出的 TF。
ERCC2、NR4A3 和其他 7 个 TF 对 PNS 对抗 IRI 具有重要意义。本研究促进了对 PNS 对抗脑 IRI 保护机制的理解，并有助于鉴定 PNS 的可能靶点。
