PMID- 34343651 OWN - NLM STAT- MEDLINE DCOM- 20220117 LR - 20220117 IS - 1872-7573 (Electronic) IS - 0378-8741 (Linking) VI - 280 DP - 2021 Nov 15 TI - A preparation of Ginkgo biloba L. leaves extract inhibits the apoptosis of hippocampal neurons in post-stroke mice via regulating the expression of Bax/Bcl-2 and Caspase-3. PG - 114481 LID - S0378-8741(21)00710-8 [pii] LID - 10.1016/j.jep.2021.114481 [doi] AB - ETHNOPHARMACOLOGICAL RELEVANCE: Shuxuening injection (SXNI) is a Chinese medicine of Ginkgo biloba L. leaves extract (GBE), which is widely used clinically for cardiovascular diseases such as stroke and myocardial infarction, but the pharmacological mechanism of its therapeutic effect is not fully understood. AIM OF THE STUDY: Preclinical studies suggested that inhibition of neuronal apoptosis effectively improves brain damage after ischemic stroke. The purpose of this study was to investigate the inhibitory effect of SXNI on neuronal apoptosis in post-stroke mice and its underlying mechanism. MATERIALS AND METHODS: A mouse cerebral ischemia-reperfusion injury (CIRI) model was constructed by middle cerebral artery occlusion (MCAO) and treated with 3 mL/kg SXNI. TUNEL and immunohistochemistry experiments were performed on brain slices on the 7th day after stroke. The protein was extracted from the hippocampus region of the brain for western-blot assay. To simulate the in vivo ischemia-reperfusion process, the hippocampal neuron cell line HT-22 was subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) in vitro, and 200 mug/mL SXNI was administered. The HT-22 cells were then studied by RT-PCR and immunocytochemistry. RESULTS: In vivo, SXNI treatment significantly reduced hippocampal neuronal apoptosis. Immunohistochemistry showed that SXNI inhibited the activation of Caspase-3 protein in the hippocampus after ischemic stroke. Western blot analysis further confirmed that SXNI regulated the expression of the antagonizing protein pair Bax and Bcl-2 to exert anti-apoptotic effect in addition to reducing the expression of Cleaved-Caspase-3 in the hippocampus. In vitro, 200 mug/mL SXNI treatment significantly improved HT-22 apoptosis caused by OGD/R. Further RT-PCR and immunocytochemistry study showed that 200 mug/mL SXNI inhibited apoptosis of hippocampal neurons by regulating the mRNA and protein expressions of apoptotic molecules Bax, Bcl-2 and Caspase-3. CONCLUSIONS: CIRI can induce hippocampal neuronal apoptosis, which is inhibited by SXNI via regulating Bax/Bcl-2 and blocking Caspase-3 activation. Therefore, SXNI may be a promising treatment strategy to improve the prognosis of ischemic stroke. CI - Copyright (c) 2021 Elsevier B.V. All rights reserved. FAU - Li, Zhixiong AU - Li Z AD - State Key Laboratory of Component-based Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology and Medicine, Tianjin, 300457, China. FAU - Xiao, Guangxu AU - Xiao G AD - State Key Laboratory of Component-based Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology and Medicine, Tianjin, 300457, China. FAU - Wang, Huanyi AU - Wang H AD - State Key Laboratory of Component-based Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology and Medicine, Tianjin, 300457, China. FAU - He, Shuang AU - He S AD - State Key Laboratory of Component-based Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology and Medicine, Tianjin, 300457, China. FAU - Zhu, Yan AU - Zhu Y AD - State Key Laboratory of Component-based Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology and Medicine, Tianjin, 300457, China. Electronic address: yanzhu.harvard@icloud.com. LA - eng PT - Journal Article DEP - 20210731 PL - Ireland TA - J Ethnopharmacol JT - Journal of ethnopharmacology JID - 7903310 RN - 0 (Bax protein, mouse) RN - 0 (Drugs, Chinese Herbal) RN - 0 (Proto-Oncogene Proteins c-bcl-2) RN - 0 (bcl-2-Associated X Protein) RN - 0 (shuxuening) RN - 114100-40-2 (Bcl2 protein, mouse) RN - EC 3.4.22.- (Casp3 protein, mouse) RN - EC 3.4.22.- (Caspase 3) SB - IM MH - Animals MH - Apoptosis/*drug effects MH - Brain Ischemia/*drug therapy MH - Caspase 3/metabolism MH - Cell Line MH - Disease Models, Animal MH - Drugs, Chinese Herbal/*pharmacology MH - Hippocampus/cytology/drug effects MH - Male MH - Mice MH - Mice, Inbred C57BL MH - Neurons/drug effects/pathology MH - Proto-Oncogene Proteins c-bcl-2/metabolism MH - Reperfusion Injury/drug therapy MH - Stroke/*drug therapy MH - bcl-2-Associated X Protein/metabolism OTO - NOTNLM OT - Ginkgo biloba L. leaves extract OT - Hippocampal neuron OT - Ischemic stroke OT - Neuronal apoptosis OT - Shuxuening injection EDAT- 2021/08/04 06:00 MHDA- 2022/01/18 06:00 CRDT- 2021/08/03 20:13 PHST- 2021/06/11 00:00 [received] PHST- 2021/07/18 00:00 [revised] PHST- 2021/07/30 00:00 [accepted] PHST- 2021/08/04 06:00 [pubmed] PHST- 2022/01/18 06:00 [medline] PHST- 2021/08/03 20:13 [entrez] AID - S0378-8741(21)00710-8 [pii] AID - 10.1016/j.jep.2021.114481 [doi] PST - ppublish SO - J Ethnopharmacol. 2021 Nov 15;280:114481. doi: 10.1016/j.jep.2021.114481. Epub 2021 Jul 31.