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Bilobalide (Bilo) alleviated oxygen-glucose-deprivation (OGD)-induced cell damage. (A) Cell activity was tested by Cell Counting Kit-8. (B-C) Cell cyclerelated proteins cyclin-dependent kinase (CDK) 4/6 and CyclinD1 were examined through western blot. (D) Cell apoptosis was measured via flow cytometry. (E-F) Expression of apoptosis relative factors was tested through western blot. Data were revealed as mean ± standard deviation (SD). Ã p < .05, ÃÃ p < .01 and ÃÃÃ p < .001 were notable consequences.
Source publication
We, the Editors and Publisher of the journal Artificial Cells, Nanomedicine, and Biotechnology, have retracted the following article:
Ailin Cao and Xiangting Li. (2019). Bilobalide protects H9c2 cell from oxygen-glucose-deprivation-caused damage through upregulation of miR-27a. Artificial Cells, Nanomedicine, and Biotechnology. 47:1, 2980–2988, DOI...
Contexts in source publication
Context 1
... in Figure 2(A) showed that, under OGD treatment for 6 h, cell activity was notably raised when bilobalide was 5 and 10 lM (both p < .05), while no significant difference was found when bilobalide was 0.1 and 1 lM (both p > .05). ...
Context 2
... bilobalide administration markedly upregulated the expression of CDK4, CDK6 and Cyclin D1 win OGD-stimulated H9c2 cells (p < .05 or p < .01, Figure 2(B,C)). On the other hand, the group treatment with OGD and bilobalide notably reduced apoptosis contrasted with OGD set (p < .05, Figure 2(D)). ...
Context 3
... the other hand, the group treatment with OGD and bilobalide notably reduced apoptosis contrasted with OGD set (p < .05, Figure 2(D)). Meanwhile, bilobalide administration also reversed the expression trend of apoptosis-related proteins presented by downregulation of p53 and Bax while Bcl-2 was upregulated (all p < .05, Figure 2(E,F)). ...
Context 4
... bilobalide administration also reversed the expression trend of apoptosis-related proteins presented by downregulation of p53 and Bax while Bcl-2 was upregulated (all p < .05, Figure 2(E,F)). Also, the rate of cleaved-Capse-3/ pro-Caspase-3 was decreased by the treatment of bilobalide (p < .01, Figure 2(E,F)). ...
Citations
... Hua et al (51) revealed that BB could protect neural cells against aggregated α-synuclein-induced apoptosis. Cao and Li (52) observed that BB ameliorated OGD-induced injury in H9c2 cells. A study by Mao et al (53) demonstrated that BB alleviated IL-17-induced inflammatory injury in ATDC5 cells. ...
Inflammation is involved in the pathological process underlying a number of liver diseases. Bilobalide (BB) is a natural compound from Ginkgo biloba leaves that was recently demonstrated to exert hepatoprotective effects by inhibiting oxidative stress in the liver cancer cell line HepG2. The anti-inflammatory activity of BB has been reported in recent studies. The major objective of the present study was to investigate whether BB could attenuate inflammation-associated cell damage. HepG2 cells were cultured with lipopolysaccharide (LPS) and BB, and cell damage was evaluated by measuring cell viability using MTT assay. The activity of the NF-κB signaling pathway was assessed by measuring the levels of IκBα, NF-κB p65, phosphorylated (p)-IκBα, p-p65, p65 DNA-binding activity and inflammatory cytokines IL-1β, IL-6 and TNF-α. A toll-like receptor (TLR)4 inhibitor (CLI-095) was used to detect the involvement of TLR4 in cell injury caused by LPS. In addition, the PI3K/Akt inhibitor LY294002 was applied to explore the involvement of the PI3K/Akt axis in mediating the effects of BB. The results demonstrated that LPS induced HepG2 cell injury. LPS also elevated the levels of p-IκBα, p-p65, p65 DNA-binding activity and inflammatory cytokines. However, CLI-095 significantly attenuated the LPS-induced cell damage and inhibited the activation of NF-κB signaling. BB also dose-dependently attenuated the LPS-induced cell damage, activation of NF-κB signaling and TLR4 overexpression. Furthermore, it was observed that LY294002 diminished the cytoprotective effects of BB on cell injury, TLR4 expression and NF-κB activation. These findings indicated that BB could attenuate LPS-induced inflammatory injury to HepG2 cells by regulating TLR4-NF-κB signaling.
... Besides, Cao et al. revealed the mitigating effects of Bilo on myocardial ischemia. Specifically, it was shown to improve oxygen-glucose deprivation (OGD)induced cell injury by enhancing miR-27a expression and activating PI3K/AKT and Wnt/β-catenin pathways [26]. We thus wondered whether Bilo alleviated OGD-induced cell injury through different mechanisms or exerted similar cardioprotective effects in vivo. ...
Myocardial infarction (MI) is an extremely severe cardiovascular disease, which ranks as the leading cause of sudden death worldwide. Studies have proved that cardiac injury following MI can cause cardiomyocyte apoptosis and myocardial fibrosis. Bilobalide (Bilo) from Ginkgo biloba leaves have been widely reported to possess excellent cardioprotective effects. However, concrete roles of Bilo in MI have not been investigated yet. We here designed both in vitro and in vivo experiments to explore the effects of Bilo on MI-induced cardiac injury and the underlying mechanisms of its action. We conducted in vitro experiments using oxygen–glucose deprivation (OGD)-treated H9c2 cells. Cell apoptosis in H9c2 cells was assessed by conducting flow cytometry assay and evaluating apoptosis-related proteins with western blotting. MI mouse model was established by performing left anterior descending artery (LAD) ligation. Cardiac function of MI mice was determined by assessing ejection fraction (EF), fractional shortening (FS), left ventricular end-systolic diameter (LVESD), and left ventricular end-diastolic diameter (LVEDD). Histological changes were analyzed, infarct size and myocardial fibrosis were measured by hematoxylin and eosin (H&E) and Masson staining in cardiac tissues from the mice. The apoptosis of cardiomyocytes in MI mice was assessed by TUNEL staining. Western blotting was applied to detect the effect of Bilo on c-Jun N-terminal kinase (JNK)/p38 mitogen-activated protein kinases (p38 MAPK) signaling both in vitro and in vivo. Bilo inhibited OGD-induced cell apoptosis and lactate dehydrogenase (LDH) release in H9c2 cells. The protein levels of p-JNK and p-p38 were significantly downregulated by Bilo treatment. SB20358 (inhibitor of p38) and SP600125 (inhibitor of JNK) suppressed OGD-induced cell apoptosis as Bilo did. In MI mouse model, Bilo improved the cardiac function and significantly reduced the infarct size and myocardial fibrosis. Bilo inhibited MI-induced cardiomyocytes apoptosis in mice. Bilo suppressed the protein levels of p-JNK and p-p38 in cardiac tissues from MI mice. Bilo alleviated OGD-induced cell apoptosis in H9c2 cells and suppressed MI-induced cardiomyocyte apoptosis and myocardial fibrosis in mice via the inactivation of JNK/p38 MAPK signaling pathways. Thus, Bilo may be an effective anti-MI agent.
... Myocardial ischemia (MI) causes fatality worldwide, and ischemic reperfusion (I/R) is the current standard strategy for its treatment. However, approximately 30 % of patients experience myocardial reperfusion (I/R)-induced injury [1]. In most cases, IR damage is due to immunity-induced accumulation of inflammatory cells in the infarction area under reperfusion. ...
Purpose: To determine the effect of γ-mangostin on myocardial ischemia (MI) -induced injury of myocardial cells, and the possible involvement of SIRT3 in myocardial cell apoptosis in Sprague-Dawley rats after ischemia-reperfusion (I/R). Methods: Ischemic reperfusion (I/R) model of rat was established, followed by TTC staining. The serum levels of CK-MB and LDH were also assessed. In addition, inflammatory response and oxidative stress were evaluated by quantitative PCR and enzyme linked immunosorbent assay (ELISA), while cell apoptosis was assessed using TdT-mediated dUTP nick end labeling (TUNEL) assay and western blot. The mechanism of action of γ-mangostin by which it mediated improvement in cardiac injury was investigated by ELISA and western blot. Results: γ-Mangostin attenuated myocardial injury and reduced myocardial inflammation in I/R rats (p < 0.05). In addition, it alleviated oxidative stress in I/R rat myocardial tissues and suppressed apoptosis. Furthermore, γ-mangostin improved myocardial injury probably by targeting SIRT3 (p < 0.05). Conclusion: γ-Mangostin has potentials for use as a therapeutic agent for the treatment of myocardial I/R injury. However, there is a need for clinical trials on the compound.
... Post-transcriptional or post-translational regulation, such as 3′UTR modification, acetylation, or methylation may be involved; miR-449a-5p [43], miR-34a-5p [44], and miR-30b-5p [45] have been found to be responsible for SIRT1-targeted inhibition. BB regulates the expression of miRNAs such as miR-101-5p and miR-27a-5p, which have multiple biological functions [46]. Synthetically, Both AMPK and SIRT1 are energy-sensing molecules that have coexisted in cells throughout evolution. ...
Background
Uncoupled extracellular matrix (ECM) causes cartilage degeneration and osteoarthritis (OA) by suppressing the synthesis and activating the degradation of ECM components. Gingko biloba is a natural Chinese herb with a variety of biological functions; however, the extent to which it can protect against OA and the mechanisms involved are unknown.
Methods
In our study, using bioinformatics tools, we were able to identify an important lactone, bilobalide (BB), from Gingko biloba . In vitro experiments were performed to evaluate the potential therapeutic effects of BB on ECM homeostasis. In vivo experiments were conducted to assess the protection of systemic administration of BB on cartilage degeneration. Molecular mechanisms underlying BB-regulated anti-arthritic role were further explored.
Results
In interleukin-1β-incubated human chondrocytes, in vitro treatment with BB increased the expression of cartilage anabolic proteins, while inhibiting the activities of ECM degrading enzymes. In a mice model, systemic administration of BB, in vivo, prevented post-traumatic cartilage erosion and attenuated the formation of abnormal osteophytes in the subchondral bone. Mechanistically, the activation of the adenosine 5′-monophosphate-activated protein kinase (AMPK)-sirtuin 1 (SIRT1) signaling pathway was involved in the anti-arthritic effects of BB. In vitro, blocking BB’s chondroprotection with the AMPK-specific inhibitor Compound C abrogated it.
Conclusions
These results demonstrated that BB extracted from Gingko biloba regulates ECM balance to prevent OA by activating the AMPK-SIRT1 signaling pathway. This study proposed the monomer BB, a traditional Chinese medicine, as a de novo therapeutic insight for OA.
Graphical Abstract
Schematic representation of the experimental design. Based on the bioinformatic analysis, bilobalide (BB), a natural herb Gingko biloba -derived ingredient, was identified as a candidate for treating osteoarthritis. In vitro, BB treatment not only facilitates cartilage extracellular matrix synthesis but also inhibits proteolytic enzyme activities. In vivo intraperitoneal injection of BB improves cartilage degeneration and subchondral bone sclerosis. BB, in particular, had anti-arthritic effects by activating the AMPK-SIRT1 signaling pathway.
... Thereby, we conducted a miRNA microarray analysis and confirmed that miR-27a was the most differentially expressed miRNAs in cells treated with ML264. Intriguingly, miR-27a has been suggested to be responsible for bilobalide's protection against oxygen-glucose-deprivation-induced damage in H9C2 cells [9]. This aroused our interests in its role in MI. ...
Myocardial infarction (MI) is a major atherosclerotic cardiovascular disease which represents a leading cause of death worldwide. Kruppel-like factor 5 (KLF5) is a member of the kruppel-like transcription factor family which has been reported with pro-apoptotic functions in myocardial cells. This work focuses on the function of KLF5 in the pathogenesis of MI and the molecules involved. A mouse model with MI was established. Hypoxia/reoxygenation (H/R)-treated H9C2 cells were applied for in vitro experiments. A KLF5-specific inhibitor ML264 was administrated in cell and animal models. ML264 significantly reduced apoptosis, expression of fibrosis-related markers, reactive oxygen species in the H/R-treated H9C2 cells, and it reduced myocardial injury, infarct size, apoptosis and fibrosis in the myocardial tissues in model mice through specific downregulation of KLF5. A microRNA (miRNA) microarray analysis was performed, which suggested miR-27a as the most upregulated miRNA in the H/R-treated cells after ML264 treatment. miR-27a mimic reduced apoptosis and fibrosis in H/R-treated cells, while miR-27a inhibition blocked the protective roles of ML264. The integrated bioinformatic analyses and luciferase assays confirmed glutamine fructose-6-phosphate transaminase 2 (GFPT2) mRNA as a target of miR-27a. Overexpression of GFPT2 counteracted the protective functions of miR-27a against MI through the activation of the TGF-β/Smad2/3 signaling pathway. To conclude, this study evidenced that KLF5 possibly induces cell and tissue damage in MI through downregulation of miR-27a and the subsequent activation of GFPT2/TGF-β/Smad2/3 axis. This study may offer novel thoughts into MI treatment.
... Rat embryonic ventricular H9c2 cardiomyocytes were maintained in DMEM medium, supplemented with a 10% (v/v) fetal bovine serum (FBS) and 1% penicillin/streptomycin, in a CO 2 incubator at 37 °C [19]. Briefly, H9c2 cells were seeded on 96-well plates at a density of 10 4 cells in each well and cultured for 24 h. ...
Noscapine is an antitumor alkaloid derived from Papaver somniferum plants. Our previous study has demonstrated that exposure of noscapine on primary murine fetal cortical neurons exposed to oxygen–glucose deprivation/reperfusion (OGD/R) has neuroprotective effects. In current study, the effects of noscapine on cardiomyocytes (H9c2 cells) damage caused by 120 minutes (min) of OGD/R were evaluated and we determined whether the addition of BD1047, sigma-one receptor antagonist, prevents the protective effects of noscapine in H9c2 cells through the production of nitric oxide (NO) and apoptosis. To initiate OGD, H9c2 cells was transferred to glucose-free DMEM, and placed in a humidified incubation chamber. Cell viability was assessed with noscapine (1–5 μM) in the presence or absence of BD1047, 24 hours (h) after OGD/R. Cell viability, NO production and apoptosis ratio were evaluated by the MTT assay, the Griess method and the quantitative real-time PCR. Noscapine considerably improved the survival of H9c2 cells compared to OGD/R. Also, noscapine was extremely capable of reducing the concentrations of NO and Bax/Bcl-2 ratio expression. While the BD1047 administration alone diminished cell viability and increased the Bax/Bcl-2 ratio and NO levels. The addition of noscapine in the presence of BD1047 did not increase the cell viability relative to noscapine alone. Noscapine exerted cardioprotective effects exposed to OGD/R-induced injury in H9c2 cells, at least partly via attenuation of NO production and Bax/Bcl-2 ratio, which indicates that the sigma-one receptor activation is involved in the protection by noscapine of H9c2 cells injured by OGD/R.
Asiaticoside is a natural triterpene compound derived from Centella asiatica, possessing confirmed cardioprotective property. However, the roles of asiaticoside in regulating oxygen-glucose deprivation/reoxygenation (OGD/R)-caused cardiomyocyte dysfunction remain largely obscure. Human cardiomyocyte AC16 cells were stimulated with OGD/R to mimic myocardial ischemia/reperfusion injury, and treated with asiaticoside. Cytotoxicity was investigated by CCK-8 assay and LDH release analysis. Autophagy- and Wnt/β-catenin signaling-related protein levels were measured via western blotting. Asiaticoside (0-20 μM) did not induce cardiomyocyte cytotoxicity. Asiaticoside (20 μM) mitigated OGD/R-induced autophagy, cytotoxicity, oxidative stress, and myocardial injury. Rapamycin, an autophagy inductor, reversed the influences of asiaticoside on autophagy, cytotoxicity, oxidative stress, and myocardial injury, while 3-methyadanine, an autophagy inhibitor, played an opposite effect. Asiaticoside (20 μM) attenuated OGD/R-induced Wnt/β-catenin signaling inactivation, which was reversed after transfection with si-β-catenin. Transfection with si-β-catenin attenuated the influences of asiaticoside on autophagy, cytotoxicity, oxidative stress, and myocardial injury. In conclusion, asiaticoside protected against OGD/R-induced cardiomyocyte cytotoxicity, oxidative stress, and myocardial injury via blunting autophagy through activating the Wnt/β-catenin signaling, indicating the therapeutic potential of asiaticoside in myocardial ischemia/reperfusion injury.
Background: The etiology of Parkinson’s disease (PD), a chronic and progressive neurodegenerative disease, is multifactorial but not fully unknown. Until now, no drug has been proven to have neuroprotective or neuroregenerative effects in patients with PD.
Objectives: To observe the therapeutic potential of Bilobalide (BB), a constituent of ginkgo biloba, in MPTP-induced PD model, and explore its possible mechanisms of action.
Material and Methods: Mice were randomly divided into three groups: healthy group, MPTP group and MPTP + BB group. PD-related phenotypes were induced by intraperitoneal injection of MPTP into male C57BL/6 mice, and BB (40 mg/kg/day) was intraperitoneally given for 7 consecutive days at the end of modeling. The injection of saline was set up as the control in a similar manner.
Results: BB induced M2 polarization of microglia, accompanied by inhibition of neuroinflammation in the brain. Simultaneously, BB promoted the expression of BDNF in astrocytes and neurons, and expression of GDNF in neurons. Most interestingly, BB enhanced the formation of GFAP⁺ astrocytes expressing nestin, Brn2 and Ki67, as well as the transformation of GFAP⁺ astrocytes expressing tyrosine hydroxylase around subventricular zone, providing experimental evidence that BB could promote the conversion of astrocytes into TH⁺ dopamine neurons in vivo and in vitro.
Conclusions: These results suggest the natural product BB may utilize multiple pathways to modify degenerative process of TH⁺ neurons, revealing an exciting opportunity for novel neuroprotective therapeutics. However, its multi-target and important mechanisms need to be further explored.
Bilobalide is a natural sesquiterpene trilactone from Ginkgo biloba leaves. It has good water solubility and is widely used in food and pharmaceutical fields. In the last decade, a plethora of studies on the pharmacological activities of bilobalide has been conducted and demonstrated that bilobalide possessed an extensive range of pharmacological activities such as neuroprotective, antioxidative, antiinflammatory, anti‐ischemic, and cardiovascular protective activities. Pharmacokinetic studies indicated that bilobalide may have the characteristics of rapid absorption, good bioavailability, wide distribution, and slow elimination. This review aims to summarize the advances in pharmacological, pharmacokinetics, toxicity, and safety studies of bilobalide in the last decade with an emphasis on its neuroprotective and antiinflammatory activities, to provide researchers with the latest information and point out the limitations of relevant research at the current stage and the aspects that should be strengthened in future research.
Full-length transcriptome sequencing based on the PacBio sequencing platform could significantly optimize the annotation of gene structures. As an ancient relic gymnosperm in the monotypic order Ginkgoales, Ginkgo biloba L. contains rich terpenoids that are medicinally valuable. The seeds have abundant edible endosperm, which is delicious and of high nutritional value. However, existing molecular studies on the developmental process of ginkgo seeds are relatively weak, and the biosynthesis of terpenoids in seeds has received little attention. Therefore, single-molecule real-time (SMRT) technology and Illumina sequencing were combined to sequence six tissues related to the reproductive growth and development of ginkgo in order to generate a high-quality full-length transcription database. In total, 20.98 Gb of clean reads containing 178,548 full-length non-chimeric (FLNC) sequences were obtained. From these data, 4019 novel genes and 22,845 novel isoforms were predicted, 52.32 % of the novel genes were annotated, and three novel isoforms were annotated in terpene synthesis related pathways. The enrichment analysis of differentially expressed genes (DEGs) showed that, 95 genes were enriched into 21 categories related to seed development, and 47 DEGs were enriched in the skeletal pathway of terpene synthesis. Combined with the real-time quantitative reverse transcription PCR (qRT-PCR), the phosphosynthase family members synthesizing terpene precursors have diverse and complex expression trends during seed development. Our findings confirm the advantages of SMRT, which facilitated the construction a rich transcript data-set for research on the development of ginkgo seeds, enriching the annotation of the ginkgo genome, and enhancing our understanding of gene regulation of terpene biosynthesis in ginkgo seeds.
