Combination Of Allicin Research Articles

Synergistic Interaction between Topical Application of Allicin and Quercetin Enhances Diabetic Wound Healing Via Inflammatory in Wistar Rats

Prolonged and excessive inflammatory processes at the wound site result in delayed healing of diabetic wounds. Allicin and quercetin have significantly inhibited tumor necrosis factor alpha (TNF-α) and increased levels of transforming growth factor beta (TGF-β) and vascular endothelial growth factor (VEGF) in rats. However, there is no known investigation on the combination of both compounds on TNF-α, TGF-β, and VEGF as a topical healing agent via inflammatory mechanisms. This study aims to determine the synergistic interaction between topical application of allicin and quercetin in enhancing diabetic wound healing via inflammatory in Wistar rats. A total of 45 male Wistar rats weighing 180-250 g were induced diabetes by a single intraperitoneal injection of streptozotocin (45 mg/kg bw). Diabetic rats underwent wound creation under anaesthesia and a square-shaped open excision wound was made with a full-thickness measurement of 1 cm × 1 cm on the right side of the rat's back. Rats were randomly allocated into group namely, vehicle control, allicin, quercetin, and a combination of allicin and quercetin which is applied topically once a day at a dose of 10 mg/mL for 7 days. Our finding showed that once daily topical application of allicin and quercetin, both independently and synergistically, increased levels of TGF-β and VEGF, as well as decreased levels of TNF-α of the rat diabetic wound model compared with the vehicle control group over 7th day. The synergistic effects of allicin and quercetin significantly enhanced diabetic wound healing via inflammation in rats, by suppressing inflammatory cytokines and stimulating growth factors hence offering a new window of an experimental study.

The combination of allicin with domiphen is effective against microbial biofilm formation.

Microorganisms in biofilms are particularly difficult to control because of their increased survival and antibiotic resistance. Allicin and domiphen were employed to inhibit the microbial growth and biofilm formation of Staphylococcus aureus, Escherichia coli, and Candida albicans strains. Broth microdilution method and checkerboard assay were conducted to determine the efficacy of allicin combined with domiphen against S. aureus, E. coli, and C. albicans. Microbial biofilm formation was measured using the crystal violet staining method and fluorescence microscopy. And the total viable count of the biofilm cells on material surface after the treatment with antimicrobial reagents was calculated with the plate count technique. The two drugs showed synergistic effects against the pathogens with a fractional bactericidal concentration of less than 0.38. The combination of 64 μg/mL allicin with 1 μg/mL domiphen dispersed over 50% of the biofilm mass of S. aureus, E. coli, and C. albicans. In addition, the drug combination reduced the total viable counts of E. coli and C. albicans biofilm cells on stainless steel and polyethylene surfaces by more than 102 CFU/mL. The combination of allicin and domiphen is an effective strategy for efficiently decreasing biofilms formation on various industrial materials surfaces.

Fractional inhibitory concentration index of the allicin and flavonoids combinations against Ganoderma boninense

Allicin and several flavonoids were known to have antifungal activity against various fungi. The antifungal capability of allicin and flavonoids varies depending on the target fungi; the combination of both compounds was expected to increase its antifungal activity. As a major disease of oil palm, basal stem rot (BSR) caused by Ganoderma boninense always be a top concern of oil palm farmers. This study aimed to evaluate the effect of combining allicin with hesperidin and rutin on antifungal activity against G. boninense through the fractional inhibitory concentration (FIC) index. The FIC index was determined using a checkerboard assay with media poisoning methods. The allicin concentration tested was 0.1%, 0.2%, 0.4%, and 0.8%, while the hesperidin and rutin concentrations were 1.25%, 2.5%, 5%, and 10%. All combinations were done in triplicate and incubated for twelve days. The result showed that a combination of allicin with hesperidin or rutin was the additive interaction that lowers the concentration of each compound with similar inhibition rate of G. boninense in vitro. Combining allicin with rutin was considered better than hesperidin due to the higher inhibition rate of G. boninense. The allicin-rutin combination has a lower concentration with a 100% inhibition rate of G. boninense in vitro.

Synergistic Antibacterial Effect and Mechanism of Allicin and an Enterobacter cloacae Bacteriophage.

Enterobacter cloacae is a troublesome pathogen causing refractory infections of the lower respiratory tract, urethra and abdominal cavity, endocarditis, osteomyelitis, and neonatal septicemia. It is prone to developing resistance to ordinary antibiotics and has brought a serious problem to clinical treatment. An artful synergistic combination of an antibacterial natural product allicin and a newly isolated bacteriophage, named BD523, was constructed herein. This combination significantly lowered effective dosage of allicin and effectually overcame bacterial drug-resistance. We experimentally evidenced that allicin interacts with bacterial DNA in the groove region by inserting itself into the DNA double helix and, subsequently, disrupts the bacterial DNA by cleaving phosphate diester bonds of deoxynucleotides. Further, BD523 destroys the cell wall and membrane of bacteria by synthesizing lyase proteins, including holin and endolysins. Thus, the synergistic effect of the combination benefits from complementary targeting mechanisms of allicin and BD523. They cooperatively act on bacterial DNA, cell wall, and membrane to improve antibacterial efficiency and avoid drug-resistance. IMPORTANCE Bacterial drug-resistance is a serious problem afflicting pharmacologists all over the world. Many strategies have been developed and practiced to overcome it, but almost no one is satisfactory due to the continual change of bacteria. Combinations of antibiotics and bacteriophages are promising because of the cooperation of 2 bacterial killers with distinct mechanisms. The combination of allicin and an Enterobacter cloacae bacteriophage reported herein can significantly improve the effect of allicin against E. cloacae. Its synergistic effect was even superior to the combination of bacteriophage and neomycin, of which the MIC was significantly lower than allicin. It was ascribed to the complementary antibacterial and the possible resistance-proof mechanism of bacteriophage and allicin. This study provided a pragmatic way to conquer the cunning bacterium, and may offer reference for research and development of new bacterial killers.

The structure, antioxidant and antibacterial properties of thiol-modified soy protein isolate induced by allicin

This study investigated the effect of allicin binding on the structure, antioxidant and antibacterial properties of soy protein isolate (SPI). Results showed that allicin bound to 82.6 % free thiol groups of SPI at a molar ratio of 0.5. The combination of allicin and SPI significantly affected the structure of protein. Result of circular dichroism showed that the content of α-helix decreased by 26.9 % and the content of β-sheet increased by 12.2 % over control when the molar ratio was 0.5. The result of surface hydrophobicity signified the unfolding of SPI with the action of allicin. These results implied that allicin binding might be a suitable method for the modification of SPI. Furthermore, the antibacterialand antioxidant experiments indicated that allicin-SPI conjugates not only had the capacity to inhibit the growth of Escherichia coli and Staphyloccocus aureus, but also had DPPH and ABTS radicals scavenging activities.

Impact of betaine and allicin against circulating oxidative and biochemical alterations induced by cadmium intoxication in common carp

This work aimed to evaluate the antidotal role of allicin and/or betaine versus experimental Cd toxicity in terms of restoring Cd-provoked changes in the circulating oxidative biomarkers together with biochemical indices of tissue abnormalities in common carp. 150 healthy common carp were subdivided into five equal groups. Group 1 fish (control) was reared in normal freshwater. Fish in other groups were exposed to 5 mg/L cadmium chloride in water for 15 days. Group 3 fish received diet supplemented with betaine (30 mg/kg BW); Group 4 fish received diet supplemented with allicin (15 mg/kg BW) and in group 5, fish food was supplemented with both betaine and allicin. Cd exposure caused significant decline in circulating SOD, catalase, GSH and FRAP as well as a rise in MDA values. Plasma levels of ALT, GGT, ALP, glucose, urea and creatinine also showed significant increase in fish treated with cadmium compared with controls. However, allicin and/or betaine supplementation had been somewhat fortunate in recovering Cd-provoked shifts in the circulating oxidative biomarkers and biochemical indicators of tissue abnormalities in common carp which may be relevant to antioxidant properties of these compounds as well as metal-chelating and methyl-donating abilities of allicin and betaine respectively. Moreover, combination of allicin and betaine stands outs as the most effective approach upon Cd poisoning. However, explanation of the precise molecular mechanisms of their modulating effects against Cd poisoning as well as their optimal dosage in various exposure durations and toxicant concentrations requires further study.

Allicin Reduces 5-fluorouracil-resistance in Gastric Cancer Cells through Modulating MDR1, DKK1, and WNT5A Expression.

5-fluorouracil (5-FU) is approved for the treatment of gastric carcinoma (GC), but chemo-resistance limits the application of it for GC. Thus, the combination of 5-FU with adjuvants such as allicin may overcome multidrug resistance (MDR). The anticancer effects of allicin, 5-FU, and allicin/5-FU on the 5-FU resistant MKN-45 cells were evaluated by MTT assay and DAPi staining. The expression of the P-glycoprotein (P-gp) and CD44 protein were determined using immunocytochemistry. We also quantified mRNA expression levels of WNT5A, Dickkopf-1 (DKK1), and MDR1 in the GC cells. Here, we found that the combination of allicin with 5-FU significantly increased apoptosis compared to 5-FU alone (P<0.05). We showed that WNT5A, MDR1, and DKK1 mRNA expression levels were down-regulated in the allicin- and allicin/5-FU-treated cells. Indeed, the combination of allicin and 5-FU significantly decreased the expression of the P-gp and CD44 proteins (P<0.05). Our findings indicate that the combination of allicin with 5-FU could reverse multidrug resistance in the GC cells by reducing the expression of WNT5A, DKK1, MDR1, P-gp, and CD44 levels.

Combinational Treatment of Imatinib and Allicin Synergistically Inhibits the Growth of BCR-ABL-Positive Leukemia and the Emergence of Drug-Resistant Cells

Imatinib mesylate (Gleevec, Glivec, STI571) is used as a first-line therapy for chronic myeloid leukemia (CML) to inhibit the tyrosine kinase activity of BCR-ABL. Although imatinib has greatly improved CML patient prognosis, imatinib-resistant cells often develop in about 30-40% of patients after treatment, causing major therapeutic failure. Several second and third generation tyrosine kinase inhibitors, such as dasatinib, nilotinib and ponatinib, have been developed, but none of them are able to completely block the development of drug-resistance. Therefore, new therapeutic approaches are in need to prevent or reduce the emergence of drug-resistant cells in the early stages of imatinib therapy.In this study, we show that the combinational treatment of allicin, an active component in garlic, and imatinib effectively blocks imatinib-resistance in K562 BCR-ABL-positive leukemia cells. Both imatinib and allicin had dose-dependent effects on cell viability during the first 3 days of treatment. However, the growth inhibitory effect did not last and resulted in re-growth of cells when treated with allicin alone (2 microgram/ml) or 200 nM of imatinib, which is close to the IC50 value in our K562 cells determined on day 3. In contrast, the combination of allicin with imatinib still inhibited cell growth by day 6. Wright-Giemsa stainings showed that the combinational treatment of imatinib and allicin also caused erythroid differentiation.To identify the molecular mechanisms of imatinib and allicin's actions, we analyzed RNA expression after single and combinational treatments with imatinib and allicin. The results demonstrated that both imatinib and allicin increased the expression of alpha -globin and gamma -globin. The expression of glycophorin A (GYPA), a marker of erythroid differentiation, was synergistically increased by the combinational treatment of imatinib and allicin. One of the notable changes we observed was a decrease of p21 (cyclin-dependent kinase inhibitor 1A; CDKN1A) expression when cells were treated with the IC50 dosage of imatinib. Surprisingly, when allicin was treated together with imatinib, the p21 decrease by imatinib was completely inhibited. Allicin alone did not increase p21, suggesting that allicin does not control p21 expression directly, but instead antagonizes the action of imatinib downregulating p21.More importantly, a long-term treatment of K562 cells with imatinib and allicin showed that allicin was able to completely block the emergence of imatinib-resistant cells in vitro. Viable cells started to grow again within 12 days when treated with imatinib or allicin alone. However, no resistant cells were observed after up to 40 days when cells were cultured with the combinational treatment.Taken together, the combinational treatment of imatinib and allicin has a synergistic effect on blocking leukemia cell expansion as they enhance erythroid differentiation and prevent cell cycle progression of residual resistant cells by attenuating the negative effects of imatinib on p21 expression. Our findings suggest that combining imatinib and allicin could be a potential therapeutic to prevent drug-resistance in BCR-ABL-positive leukemia. DisclosuresNo relevant conflicts of interest to declare.

The Synergistic Antitumor Effect of 5-Fluorouracil Combined with Allicin against Lung and Colorectal Carcinoma Cells.

5-fluorouracil (5-FU) is an anticancer drug used to inhibit the proliferation of many different tumor cells. Since severe events are associated with this compound, its combination with different anticancer drugs or adjuvants would allow the use of a significantly lower dose of 5-FU. In this study, we highlighted that the combination of allicin with 5-FU inhibited the cell migration and proliferation of colorectal and lung cancer cells. 5-FU inhibited cell growth with a similar inhibitory concentration for both normal and tumor cells (~200µM), while allicin showed different inhibitory concentrations. With an IC50 of 8.625 µM, lung cancer cells were the most sensitive to allicin. Compared to 5-FU and allicin single-agent treatments, the co-treatment showed a reduced viability rate, with p < 0.05. The morphological changes were visible on all three cell lines, indicating that the treatment inhibited the proliferation of both normal and tumor cells. We highlighted different cell death mechanisms—apoptosis for lung cancer and a non-apoptotic cell death for colorectal cancer. The synergistic antitumor effect of 5-FU combined with allicin was visible against lung and colorectal carcinoma cells. Better results were obtained when a lower concentration of 5-FU was combined with allicin than the single-agent treatment at IC50.

Does allicin combined with vitamin B-complex have superior potentials than alpha-tocopherol alone in ameliorating lead acetate-induced Purkinje cell alterations in rats? An immunohistochemical and ultrastructural study

The current article aims to explore the protective potentials of α-tocopherol alone and the combination of allicin and vitamin B-complex against lead-acetate neurotoxicity on the cerebellar cortex. Forty rats were divided into four groups (n = 10). Group 1 was the control group; Group 2 received 10 mg/kg body weight (BW) of lead acetate; Group 3 was exposed to 10 mg/kg BW of lead acetate plus a combination of allicin (100 mg/kg BW) and vitamin B-complex (40 mg/kg BW); Group 4 was administered lead acetate (10 mg/kg BW) and α-tocopherol (100 mg/kg BW). The animals received the treatment for 60 days by oral gavage. All the groups were studied ultrastructurally and immunohistochemically with glial fibrillary acidic protein (GFAP). The affected groups revealed shrunken and degenerated Purkinje cells with irregular nuclei. The cytoplasm comprised several lysosomes, unhealthy mitochondria, and dilated Golgi saccules. The myelinated nerve fibres demonstrated breaking of the myelin sheaths, apparent vacuoles, and broad axonal spaces. Immunohistochemically, there was a tremendous surge in GFAP-positive astrocytes in the lead acetate-treated group. These histological and ultrastructural variations were ameliorated by the administration of a-tocopherol and the combination of allicin and vitamin B complex. Moreover, an apparent decrease in the number of GFAP-positive astrocytes was obvious in the protected groups. Although both a-tocopherol and the combination of allicin and vitamin B-complex can be used as possible adjuvant therapies to ameliorate nervous system ailments attributable to lead acetate, α-tocopherol showed more protective potential.

Impact of allicin on enzyme activity, cytokine secretion, and gene expression dynamics in oxidatively‐stressed epithelial cells (1045.19)

Environmental stress can alter cell permeability by influencing tight junctions and inflammatory mediators. Allicin is a botanical derived from garlic that has anti‐inflammatory and antioxidant properties. The objective of the current experiment was to determine if allicin can mitigate oxidative stress using a Caco‐2 cell model. The experiment was arranged in a 3 x 2 factorial with three concentrations of allicin (0, 40, or 80 µM) and two concentrations of oxidative stressor, hydrogen peroxide (0 or 100 µM). Cells were incubated with allicin or hydrogen peroxide for 24 or 3 h, respectively. There was an increase in reactive oxygen species with hydrogen peroxide incubation, whereas a combination of allicin and hydrogen peroxide increased superoxide dismutase activity. Supplementation with allicin ameliorated the effects of hydrogen peroxide on catalase activity and reactive oxygen species. Gene expression of NFκB tended to increase and occludin expression decreased with hydrogen peroxide treatment, whereas treatment with allicin resulted in decreased expression of claudin‐4 and occludin, and an increase in IL‐8 secretion. Results from this study show that hydrogen peroxide induced oxidative damage through epithelial cell antioxidant enzymes, tight junction gene expression, and inflammatory mediators, but allicin partially mitigated epithelial cell oxidative damage.

Allicin, a SUR2 opener: possible mechanism for the treatment of diabetic hypertension in rats

The garlic (Allium sativum L., Amaryllidaceae) has been popularly used in the treatment of diabetes and cardiac complications. In the present work, we have studied the possible mechanisms, sulfonylurea receptor (SUR) selectivity of allicin in diabetic hypertensive rats. Diabetic hypertension was induced by intraperitoneal injection of streptozotocin (50 mg/kg) followed by daily administration of dexamethasone (10 µg/kg, s.c.). Different parameters, blood pressure and blood glucose levels were studied in the rats weekly up to eight weeks. Allicin (8 mg/kg, p.o.) shows potent antidiabetic (*p<0.001) as well as antihypertensive effect (**p<0.001, *p<0.01). It may act as a vasodilator by hyperpolarizing the membrane of normal vascular smooth muscle cells. The hyperpolarization in vascular smooth muscle occurs due to K+ channel opener activity. Antihypertensive effect of allicin is inhibited by glibenclamide, nonselective SUR blocker while combination of allicin with nateglinide, selective SUR1 blocker exerted synergistic antihypertensive effect. The results indicates that allicin is effective in the treatment of diabetic hypertension; through a mechanism that might involve selective opening of SUR2.

Evaluation of Novel S-Allyl Derivatives of 6-Mercapto-Purine against B-CLL: Combining the Effects of Different Compounds in A Single Molecule.

Abstract 3437Poster Board III-325Previously, we have shown that Allicin, the highly active compound of freshly crushed garlic, produced by the reaction of the enzyme Alliinase with its substrate Alliin, induced the apoptotic killing of B-CLL cells in vitro. In addition, we also reported that generation of Allicin in situ on the surface of B-CLL cells by targeting Alliinase to the cell surface of the CD20+ cells by Rituximab, resulted in the eradication of primary B-CLL in a human-mouse chimeric model, denoting the marked anti-CLL potential of combining these two different molecules, with different mechanism of action, into a single drug entity (Arditti et al., Mol Cancer Ther 2005;4(2)325-331). Indeed, monotherapeutic approaches, even if effective, are usually not sufficient to fully eradicate B-CLL and the most effective therapeutic protocols require the utilization of more than one agent. With this in mind, we took advantage of the high reactivity of Allicin, with SH-containing compounds, and created novel chimeric compounds by the combination of Allicin with 6-Mercapto-Purine (6MP) and 6MP-riboside (6MPR), both SH-containing purine analogs used for decades for the treatment of hematologic malignancies. The resulting novel compounds, S-Allyl-6MP (SA-6MP) and S-Allyl-6MPR (SA-6MPR), were examined against primary B-CLL cells obtained from the peripheral blood of patients at Binnet stage C. In our in vitro assays, Annexin-V staining indicated that SA-6MP acted in a dose dependent manner, inducing the apoptotic death of 37.9% and 95.2% of plated CD19+CD5+ B-CLL cells (10.9% in untreated cells) incubated for 16 h at 37 °C in the presence of 50 uM or 100 uM, respectively. As expected, the original 6MP compound had no impact on the viability of plated B-CLL cells (9.7% and 8.7%) at doses of up to 150 uM. In preliminary in vivo experiments, we compared the anti-BCLL activity of SA-6MP with that of SA-6MPR and the original 6MP compound on primary B-CLL cells from 5 different patients (Binnet stage C) in a human-SCID/Beige mouse model. Following the engraftment of the human B-CLL cells, mice were treated with i.p. injections of 2.5 mg/kg body weight of SA-6MP, SA-6MPR, or 6MP on a daily basis throughout 7 consecutive days, after which, the engraftment of primary B-CLL cells was examined by the recovery of CD45+CD19+CD5+ from injected mice. An additional group of mice injected with vehicle (1% DMSO) was also examined as a control. In close similarity to our in vitro results, engraftment of primary B-CLL cells was considerably reduced following treatment with SA-6MP (>90% reduction), as compared with treatment with the original 6MP drug. In addition, the chimeric riboside 6MP derivative, SA-6MPR, induced a potent anti-BCLL effect comparable to that of SA-6MP. In summary, our results in vitro and in vivo suggests that combining the pro-apoptotic effects of Allicin with the antiproliferative effects of 6MP or 6MPR is superior to the effect of either of the purine analogs alone. This approach may be evaluated at first instance in B-CLL patients with refractory disease. DisclosuresNo relevant conflicts of interest to declare.