Allicin Treatment Research Articles

Allicin Overcomes Doxorubicin Resistance of Breast Cancer Cells by Targeting the Nrf2 Pathway.

Breast cancer (BC) is a lethal disorder that threatens the life safety of the majority of females globally, with rising morbidity and mortality year by year. Doxorubicin is a cytotoxic anthracycline antibiotic that is widely used as one of the first-line chemotherapy agents for patients with BC. However, the efficacy of doxorubicin in the clinic is largely limited by its serious side effects and acquired drug resistance. Allicin (diallyl thiosulfinate), as the major component and key active compound present in freshly crushed garlic, has shown potential effects in suppressing chemotherapy resistance in various cancers. Our research aimed to explore the relationship between allicin and doxorubicin resistance in BC. To generate doxorubicin-resistant BC cell lines (MCF-7/DOX and MDA-MB-231/DOX), doxorubicin-sensitive parental cell lines MCF-7 and MDA-MB-231 were continuously exposed to stepwise increased concentrations of doxorubicin over a period of 6 months. CCK-8, colony formation, flow cytometry, RT-qPCR, and western blotting assays were performed to investigate the effects of allicin and/or doxorubicin treatment on the viability, proliferation and apoptosis and the expression of Nrf2, HO-1, phosphate AKT and AKT in doxorubicin-resistant BC cells. Our results showed that combined treatment of allicin with doxorubicin exhibited better effects on inhibiting the proliferation and enhancing the apoptosis of doxorubicin-resistant BC cells than treatment with allicin or doxorubicin alone. Mechanistically, allicin suppressed the levels of Nrf2, HO-1, and phosphate AKT in doxorubicin-resistant BC cells. Collectively, allicin improves the doxorubicin sensitivity of BC cells by inactivating the Nrf2/HO-1 signaling pathway.

Allicin regulates Sestrin2 ubiquitination to affect macrophage autophagy and senescence, thus inhibiting the growth of hepatoma cells

BackgroundAllicin regulates macrophage autophagy and senescence, and inhibits hepatoma cell growth. This study investigated the mechanism by which allicin inhibits the growth of hepatoma cells. MethodsHepa1–6 mouse hepatoma cells were subcutaneously injected into C57BL/6 J mice to construct a tumor transplantation model. Macrophages were cultured with the supernatant of hepatoma cells to construct a cell model. The levels of mRNA and proteins and the level of Sestrin2 ubiquitination were measured by RTqPCR, immunofluorescence and Western blotting. The levels of autophagy-related factors and the activity of senescence-associated β-galactosidase were determined by kits, and protein stability was detected by cycloheximide (CHX) tracking. ResultsData analysis of clinical samples revealed that RBX1 was highly expressed in tumor tissues, while Sestrin2 was expressed at low levels in tumor tissues. Allicin can promote the expression of the autophagy-related proteins LC3 and Beclin-1 in tumor macrophages and inhibit the expression of the aging-related proteins p16 and p21, thus promoting autophagy in macrophages and inhibiting cell senescence. Moreover, allicin can inhibit the expression of RBX1, thereby reducing the ubiquitination of Sestrin2, enhancing the stability of Sestrin2, activating autophagy in tumor macrophages and inhibiting senescence. In addition, allicin treatment inhibited the proliferation and migration of hepatoma carcinoma cells cocultured with macrophages and significantly improved the development of liver cancer in mice. ConclusionAllicin can affect the autophagy of macrophages and restrain the growth of hepatoma cells by regulating the ubiquitination of Sestrin2.

Potential Hepatoprotective Effects of Allicin on Carbon Tetrachloride-Induced Acute Liver Injury in Mice by Inhibiting Oxidative Stress, Inflammation, and Apoptosis.

The global burden of liver disease is enormous, which highlights the need for effective hepatoprotective agents. It was reported that allicin exhibits protective effects against a range of diseases. In this study, we further evaluated allicin's effect and mechanism in acute hepatic injury. Liver injury in mice was induced by intraperitoneal injection with 1% CCl4 (10 mL/kg/day). When the first dose was given, CCl4 was given immediately after administration of different doses of allicin (40, 20, and 10 mg/kg/day) as well as compound glycyrrhizin (CGI, 80 mg/kg/day), and then different doses of allicin (40, 20, and 10 mg/kg/day) as well as compound glycyrrhizin (CGI, 80 mg/kg/day) were administrated every 12 h. The animals were dissected 24 h after the first administration. The findings demonstrated a significant inhibition of CCl4-induced acute liver injury following allicin treatment. This inhibition was evidenced by notable reductions in serum levels of transaminases, specifically aspartate transaminase, along with mitigated histological damage to the liver. In this protective process, allicin plays the role of reducing the amounts or the expression levels of proinflammatory cytokines, IL-1β, IL-6. Furthermore, allicin recovered the activities of the antioxidant enzyme catalase (CAT) and reduced the production of malondialdehyde (MDA) in a dose-dependent manner, and also reduced liver Caspase 3, Caspase 8, and BAX to inhibit liver cell apoptosis. Further analysis showed that the administration of allicin inhibited the increased protein levels of Nuclear factor-erythroid 2-related factor 2 (Nrf2) and NAD(P)H:quinone oxidoreductase 1 (NQO1), which is related to inflammation and oxidative stress. The in vitro study of the LPS-induced RAW264.7 inflammatory cell model confirmed that allicin can inhibit important inflammation-related factors and alleviate inflammation. This research firstly clarified that allicin has a significant protective effect on CCl4-induced liver injury via inhibiting the inflammatory response and hepatocyte apoptosis, alleviating oxidative stress associated with the progress of liver damage, highlighting the potential of allicin as a hepatoprotective agent.

Allicin Improves Olfactory Response to Rewarding Stimuli Using Buried Reward Test

Garlic, known as Allium sativum, is used in food and medicine. Allicin is one of the main bioactive agents found in garlic. This study investigated the tendency for allicin, a bioactive agent in garlic, to improve olfactory response in male wistar rats exposed to cadmium chloride-induced toxicity. Thirty male wistar rats were sampled in six groups of five rats each. The treatments were; group 1 (control)- water and feed, group 2-CdCl2 (5mg/kg) group 3-allicin(20mg/k), group 4-allicin(40mg/kg), group 5- CdCl2 (5mg/kg) +allicin (20mg/kg), group 6- CdCl2 (5mg/kg) +allicin(40mg/kg). Olfactory response was tested using buried reward test with the result presented as latency (in seconds). Statistical significance was tested at 95% confidence interval (P≤0.05). There was a significant decrease (P≤0.05) in latency and an increase in olfactory response in response to increase in the dose of allicin administered. CdCl2 alone caused a significant increase (P≤0.05) in latency and decrease in olfactory response but when combined with allicin treatment there was a significant improvement in olfactory response. Allicin improves olfactory response dose-dependently using buried reward test.

ALLICIN PREVENTS MPTP‐INDUCED MOTOR DYSFUNCTION AND BIOCHEMICAL ABNORMALITIES IN WISTAR RATS

AbstractBackgroundParkinson’s disease (PD) is an age‐related progressive neurodegenerative disorder, characterized by loss of dopaminergic neurons in the substantia nigra pars compacta (SNpc) region of the brain. Dopaminergic degeneration involves the number of mechanisms such as oxidative stress and neuroinflammation. Allicin has been reported to exert various anti‐oxidative and anti‐inflammatory activities in in‐vitro and in‐vivo studies. Therefore, in the present study, we have investigated neuroprotective effect of allicin against MPTP induced Parkinson’s disease in rats.MethodRepeated intranigral infusion of MPTP at day 1st, 4th, and 7th produced significantly loss in body weight, locomotor activity, motor coordination, antioxidant defense enzymes (reduced glutathione) and significantly increase oxidative stress markers (lipid peroxidation and nitrite level) in the striatum. MPTP treatment also increased pro‐inflammatory cytokines (TNF‐α, IL‐6, and IL‐1β) level and decreased neurotransmitter level in the striatum. Allicin (5, 10 and 20mg/kg/day i.p.) treatment from day 8th to 21st significant reversed the behavioral, antioxidant defense enzymes, oxidative stress marker and pro‐inflammatory cytokines in MPTP infused rats.ResultAllicin (5, 10 and 20mg/kg/day i.p.) treatment from day 8th to 21st significant reversed the behavioral, antioxidant defense enzymes, oxidative stress marker and pro‐inflammatory cytokines in MPTP infused rats.ConclusionThe consequences of particular study suggest the beneficial effects of allicin in PD.

Antioxidant and anti-inflammatory effects of allicin in the kidney of an experimental model of metabolic syndrome.

Recent studies have suggested that metabolic syndrome (MS) encompasses a group of risk factors for developing chronic kidney disease (CKD). This work aimed to evaluate the antioxidant and anti-inflammatory effects of allicin in the kidney from an experimental model of MS. Male Wistar rats (220-250 g) were used, and three experimental groups (n = 6) were formed: control (C), metabolic syndrome (MS), and MS treated with allicin (16 mg/Kg/day, gastric gavage) (MS+A). MS was considered when an increase of 20% in at least three parameters (body weight, systolic blood pressure (SBP), fasting blood glucose (FBG), or dyslipidemia) was observed compared to the C group. After the MS diagnosis, allicin was administered for 30 days. Before the treatment with allicin, the MS group showed more significant body weight gain, increased SBP, and FBG, glucose intolerance, and dyslipidemia. In addition, increased markers of kidney damage in urine and blood. Moreover, the MS increased oxidative stress and inflammation in the kidney compared to group C. The allicin treatment prevented further weight gain, reduced SBP, FBG, glucose intolerance, and dyslipidemia. Also, markers of kidney damage in urine and blood were decreased. Further, the oxidative stress and inflammation were decreased in the renal cortex of the MS+A compared to the MS group. Allicin exerts its beneficial effects on the metabolic syndrome by considerably reducing systemic and renal inflammation as well as the oxidative stress. These effects were mediated through the Nrf2 pathway. The results suggest allicin may be a therapeutic alternative for treating kidney injury induced by the metabolic syndrome risk factors.

Allicin ameliorates sepsis-induced acute kidney injury through Nrf2/HO-1 signaling pathway.

Acute kidney injury (AKI) is a complication that can be induced by different factors. Allicin is a class of organic sulfur compounds with anticancer and antibacterial effects, and has not been reported in sepsis-induced AKI (S-AKI). S-AKI was induced in c57BL/6 mice by cecal ligation puncture. In response to the treatment of allicin, the survival rate of mice with S-AKI was increased. Reduced levels of serum creatinine, blood urea nitrogen, UALB, KIM-1 and NGAL indicated an improvement in renal function of S-AKI mice. Allicin inhibited the inflammation and cell apoptosis, which evidenced by decreased levels of inflammatory cytokines and apoptosis-related proteins. Oxidative stress was evaluated by the levels of oxidative stress biomarkers, and suppressed by allicin. In addition, allicin-alleviated mitochondrial dysfunction was characterized by decreased JC-1 green monomer. These effects of allicin were also evidenced in HK2 cells primed with lipopolysaccharide (LPS). Both in vivo and in vitro experiments showed that the nuclear translocation of Nrf2 and the expression of HO-1 increased after allicin treatment, which was confirmed by ML385 and CDDO-Me. In summary, this study revealed the alleviating effect of allicin on S-AKI and demonstrated the promotive effect of allicin on nuclear translocation of Nrf2 for the first time. It was inferred that allicin inhibited the progression of S-AKI through Nrf2/HO-1 signaling pathway. This study makes contributions to the understanding of the roles of allicin in S-AKI.

Evaluation of antiepileptic activity of ethanolic extract of garlic containing 3.25% allicin in experimental animals

IntroductionEpilepsy is a condition of the brain that causes seizures and affects a significant portion of the global population, and current treatments often have limited efficacy and undesirable side effects. Throughout history, people have utilized garlic for its medicinal properties to address diverse health concerns. Allicin is a major component of garlic. We conducted an experiment to evaluate the antiepileptic properties of an ethanolic extract of garlic (EEG) with 3.25% allicin content using acute epilepsy models. MethodsFor our experiment, we employed 72 Swiss albino mice, which were divided into six separate groups. Chemo-convulsant PTZ and strychnine HCL were used to induce epileptic activity in the mice, and the standard used was diazepam (5 mg/kg p.o.). Our study evaluated the effectiveness of an ethanolic garlic extract (EEG) with the same potency as allicin at doses of 25, 50, and 100 mg/kg (p.o.). on epileptic activity was assessed using a variety of parameters. The GABA level of mice brains was also estimated. ResultsThe allicin treatment led to a notable decrease in the duration and increase in the latency of seizures that were induced by PTZ and strychnine. The extract also showed a significant decrease in the mortality rate. Test allicin intervention restored brain GABA levels in a dose-dependent manner. ConclusionOur study indicates that the EEG containing 3.25% allicin has the potential as an antiepileptic agent, possibly due to its modulation of the GABAergic neurotransmitter systems.

Synergistic effect of ultrahigh pressure and allicin on gel properties, flavor characteristics, and myosin structure of the obturator muscle of the scallop (Patinopecten yessoensis).

The synergistic effects of the combination of ultrahigh pressure (UHP) with allicin on the gel properties, flavor characteristics, and myosin structure of scallops were investigated. The results indicated that chewiness reached maximum, uniform, and dense microstructures at B-300MPa, and scallops with favorable gel properties. In addition, the electronic nose and tongue could clearly distinguish the olfactory and gustatory properties of scallops, and the interaction of UHP and allicin increased the variety of volatile compounds in scallops, which mainly included 1-hydroxy-2-propanone, 1-hexenal, 2-butanone-D, and 1-octen-3-ol. The main performance was fruit aroma and a plantlike aroma and mushroomlike odor. UHP and allicin changed the microenvironment of tryptophan residues, and allicin formed larger aggregates by forming disulfides with myosin. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis results could show that myosin had low degradation in B-300MPa. Thus, comprehensively viewed, UHP and allicin play a role in gel formation of myosin from obturator muscle at 300MPa, whereas allicin and myosin form disulfides as the main factor of myosin gelation. PRACTICAL APPLICATION: To enhance the diversity of scallop preparation methods and improve the quality of the obtained product, UHP and allicin treatment result in scallops with satisfactory chewiness and flavor, which provides application prospects for scallop processing.

Antifungal Activity and Potential Action Mechanism of Allicin against Trichosporon asahii.

Trichosporon asahii is an emerging opportunistic pathogen that causes potentially fatal disseminated trichosporonosis. The global prevalence of coronavirus disease 2019 (COVID-19) poses an increasing fungal infection burden caused by T. asahii. Allicin is the main biologically active component with broad-spectrum antimicrobial activity in garlic. In this study, we performed an in-depth analysis of the antifungal characteristics of allicin against T. asahii based on physiological, cytological, and transcriptomic assessments. In vitro, allicin inhibited the growth of T. asahii planktonic cells and biofilm cells significantly. In vivo, allicin improved the mean survival time of mice with systemic trichosporonosis and reduced tissue fungal burden. Electron microscopy observations clearly demonstrated damage to T. asahii cell morphology and ultrastructure caused by allicin. Furthermore, allicin increased intracellular reactive oxygen species (ROS) accumulation, leading to oxidative stress damage in T. asahii cells. Transcriptome analysis showed that allicin treatment disturbed the biosynthesis of cell membrane and cell wall, glucose catabolism, and oxidative stress. The overexpression of multiple antioxidant enzymes and transporters may also place an additional burden on cells, causing them to collapse. Our findings shed new light on the potential of allicin as an alternative treatment strategy for trichosporonosis. IMPORTANCE Systemic infection caused by T. asahii has recently been recognized as an important cause of mortality in hospitalized COVID-19 patients. Invasive trichosporonosis remains a significant challenge for clinicians, due to the limited therapeutic options. The present work suggests that allicin holds great potential as a therapeutic candidate for T. asahii infection. Allicin demonstrated potent in vitro antifungal activity and potential in vivo protective effects. In addition, transcriptome sequencing provided valuable insights into the antifungal effects of allicin.

Allicin Inhibits Porcine Reproductive and Respiratory Syndrome Virus Infection In Vitro and Alleviates Inflammatory Responses.

Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most economically important pathogens to the swine industry worldwide over the past three decades. No approved effective antiviral drug is available to control this virus. The antiviral effects of allicin (diallyl thiosulfinate) on many human and animal viruses have been documented. However, the antiviral effect of allicin on PRRSV infection remains unknown. In this study, we found that allicin exhibited an inhibitory effect on HP-PRRSV and NADC30-like PRRSV in a dose-dependent manner by interfering with viral entry, replication, and assembly. Furthermore, allicin alleviated the expression of pro-inflammatory cytokines (IFN-β, IL-6, and TNFα) induced by PRRSV infection. The pro-inflammatory signaling pathways, TNF signaling pathway and MAPK signaling pathway, up-regulated by PRRSV infection were restored by allicin treatment. Taken together, these results demonstrate that allicin has antiviral activity against PRRSV and ameliorates inflammatory responses induced by PRRSV infection, suggesting that allicin is a promising drug candidate for anti-PRRSV therapy in vivo.

The protective roles of allicin on type 1 diabetes mellitus through AMPK/mTOR mediated autophagy pathway.

Background: Type 1 diabetes mellitus (T1DM) is one of the most common endocrine and metabolic diseases in children. Pancreatic β cells are thought to be critical cells involved in the progression of T1DM, and their injury would directly lead to impaired insulin secretion. Purpose: To investigate the protective effects of allicin on pancreatic β cell injury and elucidate the underlying mechanism. Methods: The streptozotocin (STZ)-induced mouse T1DM model in vivo and STZ-induced pancreatic β cell Min6 model in vitro were used to explore the effects of allicin on T1DM. The experiments include fasting blood glucose test, oral glucose tolerance detection, HE staining, immunohistochemistry, immunofluorescence, TUNEL staining, western blot, real-time quantitative PCR (RT-qPCR), and flow cytometry. Results: Allicin could significantly decrease blood glucose level, improve islet structure and insulin expression, and inhibit apoptosis to reduce STZ-induced pancreatic β cell injury and loss through activating AMPK/mTOR mediated autophagy pathway. Conclusion: Allicin treatment significantly reduced STZ-induced T1DM progression, suggesting that allicin may be a potential therapy option for T1DM patients.

Allicin Alleviates Diabetes Mellitus by Inhibiting the Formation of Advanced Glycation End Products.

Advanced glycation end products (AGEs) cause damage to pancreatic β-cells and trigger oxidative stress and inflammation, which promotes the development and progression of diabetes and its complications. Therefore, it is important to inhibit the formation of AGEs as part of the treatment of diabetes. Allicin is a natural antimicrobial agent with abundant pharmacological activities, and recent studies have reported its therapeutic effects in diabetes; however, the mechanism of these therapeutic effects is still unclear. Thus, the purpose of this study was to further investigate the association between allicin treatment of diabetes and AGEs. First, we established a streptozocin (STZ)-induced diabetic rat model and treated the rats with allicin for six weeks. We measured glycolipid metabolism, AGE levels, receptor of advanced glycation end products (RAGE) levels, oxidative stress, and other related indicators. The results showed that allicin improved blood glucose and body weight, reduced lipid accumulation, and inhibited AGE formation in rats. Treatment with allicin also inhibited RAGEs and thereby prevented AGE activity, which, in turn, alleviated oxidative stress and promoted insulin secretion. To further verify the effect of allicin on AGEs, we also performed in vitro nonenzymatic glycation simulation experiments. These results showed that allicin inhibited the production of AGEs by suppressing the production of AGEs intermediates. Thus, our research suggests that allicin may alleviate diabetes by inhibiting the formation of AGEs and reducing RAGE levels to relieve oxidative stress and promote insulin secretion.

RETRACTED: Allicin promotes autophagy and ferroptosis in esophageal squamous cell carcinoma by activating AMPK/mTOR signaling

The antitumor effects of allicin have been demonstrated in various cancers. However, whether allicin improves esophageal squamous cell carcinoma (ESCC) has not yet been explored. The present study aimed to explore the function and underlying mechanism of action of allicin in ESCC treatment. Our data showed that allicin significantly suppressed ESCC cell proliferation in a dose- and time-dependent manner. A green fluorescent protein-light chain 3 (LC3) transfection assay showed that autophagosomes were elevated in ESCC cells treated with allicin compared with control ESCC cells and that 3-methyladenine (an autophagy inhibitor) reversed allicin-induced LC3 puncta. Furthermore, allicin significantly elevated the ratio of LC3II/LC3I but decreased p62 expression in ESCC cells. Allicin also increased adenosine monophosphate-activated protein kinase (AMPK) phosphorylation but decreased that of the mechanistic target of rapamycin kinase (mTOR), which then induced the elevation of autophagy-related 5 and autophagy-related 7 proteins in ESCC cells. Furthermore, allicin treatment increased the expression of nuclear receptor coactivator 4 (a selective cargo receptor) but suppressed the expression of ferritin heavy chain 1 (the major intracellular iron-storage protein) in ESCC cells and elevated malondialdehyde and Fe2+ production levels. In vivo assays showed that allicin significantly decreased tumor weight and volume. In summary, allicin may induce cell death in ESCC cells by activating AMPK/mTOR-mediated autophagy and ferroptosis. Therefore, allicin may have excellent potential for use in the treatment of ESCC.

Effect of Allicin and Artesunate Combination Treatment on Experimental Mice Infected with Plasmodium berghei.

Malaria is still a significant health problem in endemic countries and increases Plasmodium resistance to the available antimalarial drugs. Hence, this study aimed to investigate the antimalarial activity of allicin and its combination with artesunate (ART) against rodent malaria Plasmodium berghei ANKA (PbANKA) infected mice. Allicin was prepared in 20% Tween-80. Balb/c mice were inoculated intraperitoneally with 1×107 PbANKA-infected erythrocytes and orally given by gavage with the chosen doses of 1, 10, 50, and 100 mg/kg of allicin and 1, 5, 10, and 20 mg/kg of ART once a day for 4 consecutive days. Effective dose 50 (ED50) of allicin and ART was subsequently investigated. Moreover, the combination (1 : 1) of allicin and ART at the doses of their respective ED50, ED50 1/2, ED50 1/4, and ED50 1/8 was also carried out. The untreated control was given 20% Tween-80. The results showed that allicin presented a dose-dependent antimalarial activity with significance (p < 0.05). The ED50 values of allicin and ART were about 14 and 5 mg/kg, respectively. For combination, allicin and ART showed a synergistic effect at the combination doses of ED50, ED50 1/2, and ED50 1/4 with significantly (p < 0.01) prevented reduction of packed cell volume, bodyweight loss, rapid dropping of rectal temperature, and markedly prolonged mean survival time, compared with the untreated control and single treatment. It can be concluded that allicin exerted potential antimalarial activity in single and its combination with ART.

In vitro neuroprotective effects of allicin on Alzheimer’s disease model of neuroblastoma cell line

Background/Aim: Alzheimer's disease is a progressive disorder that causes atrophy and neuronal death in the brain. Currently, there is not any effective therapy for Alzheimer's disease. The current research was designed to investigate the beneficial effects of allicin on Alzheimer’s disease in SHSY-5Y cells in vitro and elucidating the neuroprotective mechanism of allicin. Methods: Human neuroblastoma cell line (SH-SY5Y) was differentiated with retinoic acid to conduct the in vitro Alzheimer’s Disease model. Amyloid β1-42 protein was applied to the cells for 24 hours (2.5 µM) to induce cytotoxicity. Allicin was applied to the cell cultures in a wide spectrum dose (10 μM, 50 μM, 100 μM) to investigate neuroprotective effect against amyloid β for 24 hours. MTT and LDH analyses were performed to assess the cell viability. MDA and ROS concentrations and SOD activity were analyzed to determine the oxidative stress. Moreover, the effects of allicin on the caspase-3 expression in amyloid β induced neurotoxicity were determined by the RT-PCR analysis. Results: Amyloid β markedly decreased cell viability of SH-SY5Y in MTT analysis and elevated LDH levels. In contrast, in MTT analysis, the allicin markedly increased cell viability, indicating that allicin induces cell proliferation. Moreover, in LDH analysis, allicin treatment markedly decreased LDH release. Exposure to amyloid β markedly increased MDA and ROS levels, in comparison with the control. Moreover, amyloid β decreased activity of SOD in SH-SY5Y cells. Allicin markedly balanced out the amyloid β-induced MDA and ROS generation. In the same pathway, allicin increased activity of SOD in amyloid β -exposed SH-SY5Y cells. The caspase 3 expression was increased in amyloid β group in comparison to the control group and allicin markedly lowered the expression of caspase-3 levels. Conclusion: The beneficial effects of allicin on amyloid β -induced neurotoxicity on SH-SY5Y cells were reported for the first time in terms of cell viability, oxidative stress and apoptosis.

The Effect of Allicin on the Proteome of SARS-CoV-2 Infected Calu-3 Cells

Allicin (diallyl thiosulfinate) is the major thiol-reactive organosulfur compound produced by garlic plants (Allium sativum) upon tissue damage. Allicin exerts its strong antimicrobial activity against bacteria and fungi via S-thioallylation of protein thiols and low molecular weight thiols. Here, we investigated the effect of allicin on SARS-CoV-2 infected Vero E6 and Calu-3 cells. Toxicity tests revealed that Calu-3 cells showed greater allicin tolerance, probably due to >4-fold higher GSH levels compared to the very sensitive Vero E6 cells. Exposure of infected Vero E6 and Calu-3 cells to biocompatible allicin doses led to a ∼60–70% decrease of viral RNA and infectious viral particles. Label-free quantitative proteomics was used to investigate the changes in the Calu-3 proteome after SARS-CoV-2 infection and the effect of allicin on the host-virus proteome. SARS-CoV-2 infection of Calu-3 cells caused a strong induction of the antiviral interferon-stimulated gene (ISG) signature, including several antiviral effectors, such as cGAS, Mx1, IFIT, IFIH, IFI16, IFI44, OAS, and ISG15, pathways of vesicular transport, tight junctions (KIF5A/B/C, OSBPL2, CLTCL1, and ARHGAP17) and ubiquitin modification (UBE2L3/5), as well as reprogramming of host metabolism, transcription and translation. Allicin treatment of infected Calu-3 cells reduced the expression of IFN signaling pathways and ISG effectors and reverted several host pathways to levels of uninfected cells. Allicin further reduced the abundance of the structural viral proteins N, M, S and ORF3 in the host-virus proteome. In conclusion, our data demonstrate the antiviral and immunomodulatory activity of biocompatible doses of allicin in SARS-CoV-2-infected cell cultures. Future drug research should be directed to exploit the thiol-reactivity of allicin derivatives with increased stability and lower human cell toxicity as antiviral lead compounds.

Allicin protects against myocardial I/R by accelerating angiogenesis via the miR-19a-3p/PI3K/AKT axis.

Objectives: Allicin is an allyl 2-propenethiosulfinate or diallyl thiosulfinate acid with cardioprotective effects in myocardial ischemia/reperfusion (MI/R) injury. This study aims to examine the underlying mechanism by which Allicin protects against MI/R.Methods: C57BL6 mice were subjected to either sham or MI/R surgery, and mice in the Allicin group were injected with Allicin (5 mg/ml) before the induction of ischemia. The cardiac function and histopathology of experimental mice were evaluated by ultrasound quantification and Masson staining. We next measured the capillary angiogenesis of the peri-infarct area by Masson staining and immunohistochemical staining. The miRNA microarray was carried out to examine the expressed miRNAs in MI/R tissues and corresponding normal tissues. Real-time quantitative polymerase chain reaction (q-PCR) was performed to validate the selected miRNA-19α-3p gene expression. Besides, we evaluated the myocardial lactate dehydrogenase and COX-2 by immunofluorescence staining. The western blot analysis was used to evaluate the protein levels of p-AKT, p-PI3K, p-mTOR, COX-2, and VEGF protein in the Allicin and Model group. In vitro study, LPS stimulated Tie2 expressing macrophages were cultured in an ischemic buffer. We evaluated the accumulation of VEGF by fura-2/AM fluorescence. Besides, Western blotting was performed to examine the protein levels of p-PI3K, p-AKT, p-mTOR, VEGF, COX2, and MMP2. The PI3K inhibitor was applied to investigate whether Allicin-induced myocardial ischemia-reperfusion injury protection is mediated via the PI3K/AKT pathway. And the miR-19α-3p mimic/inhibitor were transfected to promote/inhibit the expression of miR-19a-3p for verifying the regulation of miR-19a-3p on PI3K pathway.Results: Allicin pretreatment significantly improved I/R-induced cardiac function damage. Furthermore, Allicin could repress cardiac fibrosis, as evidenced by reduced areas of cardiac fibrosis. Allicin’s effect on the MI/R was associated with increased capillary angiogenesis. Microarray analysis exposed that miR-19a-3p down-regulated PIK3CA (PI3K) expression by directly targeting the PIK3CA gene. The regulation of the angiogenesis pathway and gene miRNA-19a-3p might affect the Allicin-induced MI/R protection. Immunofluorescence staining revealed that COX-2 and myocardial lactate dehydrogenase were significantly increased after Allicin treatment. Furthermore, western blot analysis demonstrated that p-AKT, p-PI3K, p-mTOR, COX-2, and VEGF protein levels were also increased in the Allicin group. In vitro study, the protein levels of p-PI3K, p-AKT, p-mTOR, VEGF, COX2, and MMP2 were significantly increased in the Allicin-treated Tie2 expressing macrophages. These effects were partially reversed by PI3K inhibitor (Wortmannin) treatment. MiR-19α-3p plays an important role in myocardial I/R injury. It could regulate the activity of the PI3K-AKT pathway. And inhibition of miR-19a-3p promoted angiogenesis by regulating PI3K/AKT pathway.Conclusions: Allicin pretreatment protects against myocardial I/R and activating the miR-19a-3p/PI3K/AKT pathway.

Ameliorative effect of allicin on vascular calcification via inhibiting endoplasmic reticulum stress.

Vascular calcification (VC) is an independent predictor for cardiovascular events and mortality. However, there are currently no effective methods to reverse or prevent it. The present study aimed to determine the ameliorative effect of allicin on VC. VC model of rats was induced by high-dose vitamin D3, which was valued by Alizarin Red staining, calcium contents, and alkaline phosphatase in the aorta. Systolic blood pressure, pulse pressure, and pulse wave velocity were measured to determine aortic stiffness. Protein levels were detected by Western blot. Allicin treatment rescued aortic VC and stiffness. The increased protein levels of RUNX2 and BMP2, two markers of osteoblastic phenotype of vascular smooth muscle cells, in the calcified aorta were attenuated by allicin, whereas the decreased levels of calponin and SM22α induced by calcification were improved. Allicin treatment significantly attenuated the increased protein levels of GRP78, GRP94, and CHOP, which are key markers of endoplasmic reticulum stress, in the calcified aorta. The activation of PERK/eIF2α/ATF4 cascades was also prevented by allicin. Allicin could ameliorate aortic VC and stiffness. The ameliorative effect of allicin on VC might be mediated by inhibiting PERK/eIF2α/ATF4 cascades. Our results might provide a new proof for VC treatment.

Evaluation of Allicin Against Alveolar Echinococcosis In Vitro and in a Mouse Model

PurposeAt present, the chemotherapy for alveolar echinococcosis (AE) is mainly based on albendazole (ABZ). However, more than 20% of patients fail chemotherapy. Therefore, new and more effective treatments are urgently needed. Allicin has been reported to have antibacterial and antiparasitic effects. The objectives of the present study were to investigate the in vivo and in vitro efficacy of allicin against Echinococcus multilocularis (E. multilocularis).MethodsThe effects of allicin on protoscolex survival and structural changes were evaluated in vitro. The 4-week-old BALB/c male mice used for in vivo modelling underwent inoculation of E. multilocularis protoscoleces by intraperitoneal injection, followed by intragastric administration of allicin for 6 weeks. Then, the effects of allicin on lymphocyte subsets, metacestode growth and host tissue matrix metalloproteinase 2 (MMP2)/MMP9 expression around metacestodes in mice were evaluated. The toxicity of allicin was further evaluated in vivo and in vitro.ResultsAtt 40 μg/mL, allicin showed a killing effect on protoscoleces in vitro and treatment resulted in the destruction of protoscolex structure. Molecular docking showed that allicin could form hydrogen bonds with E. multilocularis cysteine enzymes. After 6 weeks of in vivo allicin treatment, the spleen index of mice was increased and the weight of metacestodes was reduced. Allicin increased the proportion of CD4+ T cells and decreased the proportion of CD8+ T cells in the peripheral blood and spleen. Pathological analysis of the metacestodes showed structural disruption of the germinal and laminated layers after allicin treatment. In addition, allicin inhibited the expression of MMP2 and MMP9 in metacestode-surrounding host tissues. At 160 μg/mL, allicin had no significant toxicity to normal hepatocytes but could inhibit hepatoma cell proliferation. At 30 mg/kg, allicin had no significant hepatorenal toxicity in vivo.ConclusionThese results suggest that allicin exerts anti-E. multilocularis effects in vitro and in vivo and can enhance immune function in mice, with the potential to be developed as a lead compound against echinococcosis.