title: Effects of the Combination of the Main Active Components of Astragalus and Panax notoginseng on Inflammation and Apoptosis of Nerve Cell after Cerebral Ischemia-Reperfusion.
abstract: Astragalus and Panax notoginseng are commonly used to treat cardio-cerebrovascular diseases in China and are often combined together to promote curative effect. We speculate that the enhancement of the combination on anticerebral ischemia injury may come from the main active components. The purpose of this work was to probe the effects and mechanisms of Astragaloside IV (the active component of Astragalus) combined with Ginsenoside Rg1, Ginsenoside Rb1, and Notoginsenoside R1 (the active components of P. notoginseng) to antagonize ischemia/reperfusion (I/R) injury via inflammation and apoptosis. C57BL/6 mice were randomly divided into sham, model, Astragaloside IV, Ginsenoside Rg1, Ginsenoside Rb1, Notoginsenoside R1, four active components combination, and Edaravone groups. After administration for 3 days, bilateral common carotid arteries (CCA) were occluded with artery clip for 20[Formula: see text]min followed by reperfusion for 24[Formula: see text]h. Our results showed that the survival rate of nerve cell in hippocampal CA1 decreased while the apoptotic rate increased, and the level of caspase-3 protein in brain tissues was elevated, the expressions of TNF-a, IL-1, and ICAM-1 mRNA as well as phosphorylated nuclear factor kappa B (NF-κB) inhibitor protein α (p-IκBa) in brain tissues were up-regulated, and the nuclear translocation rate of NF-κB was raised. Additionally, the protein expressions of phosphorylated tyrosine kinase 1 (p-JAK1), phosphorylated signal transducer and activator of transcription-1 (p-STAT1), glucose regulated protein 78 (GRP78), caspase-12, and phosphorylated c-Jun N-terminal kinases 1/2 (p-JNK1/2) in brain tissues were also significantly strengthened after I/R for 24 h. All drugs could increase neurocyte survival rate in hippocampal CA1, decrease the apoptotic rate, and inhibit caspase-3 protein expression, in contrast, the effects of four active components combination were better than those of active components alone. In addition, Astragaloside IV and Ginsenoside Rg1 could down-regulate the level of TNF-α, and ICAM-1 mRNA, respectively, Notoginsenoside R1 reduced both TNF-α and ICAM-1 mRNA, and the combination of the 4 effective components had inhibitory effects on the expressions of TNF-α, IL-1β, and ICAM-1 mRNA. Astragaloside IV, Ginsenoside Rg1, Notoginsenoside R1, and 4 effective components combination were able to restrain the phosphorylation of IκBα, and relieve the nuclear translocation rate of NF-κB. Moreover, the effects of the combination are greater than those of active components alone. All drugs could suppress the phosphorylation of JAK1 induced by I/R; meanwhile the expression of p-STAT1 exhibited a decrease in Ginsenoside Rg1 and four active components combination groups. The decreases of p-JAK1 and p-STAT1 in the four active components combination group were more obvious than those in active components alone groups. Astragaloside IV, Ginsenoside Rg1, and Notoginsenoside R1 further augmented GRP78 expression caused by I/R, Notoginsenoside R1 attenuated caspase-12 protein expression, Astragaloside IV and Ginsenoside Rg1 lessened the phosphorylation of JNK1/2, and the four active components combination was capable of up-regulating GRP78 protein while down-regulating the expressions of caspase-12 and p-JNK1/2. Similarly, the effects of the four active components combination were greater than those of effective components alone. These suggested that the combination of the main active components of Astragalus and Panax notoginseng could strengthen protective effects on cerebral ischemia injury via anti-apoptosis and anti-inflammation, and the mechanisms might be associated with restraining the activation of NF-κB and JAK1/STAT1 signal pathways and regulating endoplasmic reticulum stress (ERS) after cerebral ischemia.
authors.affiliation: * Molecular Pathology Laboratory, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, P.R. China.
authors.forename: Xiao-Ping
authors.initials: XP
authors.lastname: Huang
authors.affiliation: † Key Laboratory of Hunan Province for Prevention and Treatment of Integrated, Traditional Chinese and Western Medicine on Cardio-Cerebral Diseases, Changsha 410208, Hunan Province, P.R. China.
authors.forename: Huang
authors.initials: H
authors.lastname: Ding
authors.affiliation: ‡ Key Laboratory of Hunan Universities for Cell biology and Molecular Techniques, Changsha 410208, Hunan Province, P.R. China.
authors.forename: Jin-Dong
authors.initials: JD
authors.lastname: Lu
authors.affiliation: † Key Laboratory of Hunan Province for Prevention and Treatment of Integrated, Traditional Chinese and Western Medicine on Cardio-Cerebral Diseases, Changsha 410208, Hunan Province, P.R. China.
authors.forename: Ying-Hong
authors.initials: YH
authors.lastname: Tang
authors.affiliation: ‡ Key Laboratory of Hunan Universities for Cell biology and Molecular Techniques, Changsha 410208, Hunan Province, P.R. China.
authors.forename: Bing-Xiang
authors.initials: BX
authors.lastname: Deng
authors.affiliation: * Molecular Pathology Laboratory, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, P.R. China.
authors.forename: Chang-Qing
authors.initials: CQ
authors.lastname: Deng
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics; D027883:Astragalus Plant / Q000737:chemistry*; D002545:Brain Ischemia / Q000188:drug therapy* / Q000473:pathology*; D056547:CA1 Region, Hippocampal / Q000166:cytology; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D004195:Disease Models, Animal; D004338:Drug Combinations; D000091342:Endoplasmic Reticulum Chaperone BiP; D036145:Ginsenosides / Q000008:administration & dosage* / Q000302:isolation & purification / Q000494:pharmacology; D007249:Inflammation / Q000188:drug therapy* / Q000473:pathology*; D053613:Janus Kinase 1; D008297:Male; D008810:Mice, Inbred C57BL; D016328:NF-kappa B; D009474:Neurons / Q000473:pathology*; D018696:Neuroprotective Agents; D053723:Panax notoginseng / Q000737:chemistry*; D008517:Phytotherapy; D015427:Reperfusion Injury / Q000188:drug therapy* / Q000473:pathology*; D050794:STAT1 Transcription Factor; D012503:Saponins / Q000008:administration & dosage* / Q000302:isolation & purification / Q000494:pharmacology; D014315:Triterpenes / Q000008:administration & dosage* / Q000302:isolation & purification / Q000494:pharmacology
chemical_list: D004338:Drug Combinations; D000091342:Endoplasmic Reticulum Chaperone BiP; D036145:Ginsenosides; C000717248:Hspa5 protein, mouse; D016328:NF-kappa B; D018696:Neuroprotective Agents; D050794:STAT1 Transcription Factor; D012503:Saponins; C494080:Stat1 protein, mouse; D014315:Triterpenes; C052064:astragaloside A; D053613:Janus Kinase 1; C072936:notoginsenoside R1
keywords: Active Component Combination; Apoptosis; Astragalus; Cerebral I/R; Inflammation; Panax notoginseng