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Objective:To investigate the antibacterial efficacy of allicin effective component,dially trisulfide(DATS),on the biofilms of Enterococcus faecalis(E.faecalis)in vitro.Methods:60 healthy single root canal permanent teeth were selected.After cleaning and autoclaving,15 teeth without infection were used in the negative control group,45 teeth were contaminated by E.faecalis and incubated for 28 days as the infection models and devided into 3 groups(n=15)as below:DATS group,Calcium hydroxide group and positive(without antibacteria treatment)control group.The minial inhibition concentration(MIC)and minial bactericidal concentrition(MBC)of DATS on E.feacalis were studied with doubling diluting method.At the day 1,2 and 7 of the exprement,the 4 mg of root canal in-ner dentin powder,5 specimens of each group were sespectively transfered into sterile bottle containing 2 mL BHI broth and cultured at 37 ℃ 5%CO2 for 72 h,the turbitity of the upper layer of liquid of the culture was measured.Results:MIC and MBC(μg/mL)of DATS to E.feacalis was 2 560 and 5 120 repsectively.At the first day,turbidity in the DATS group was the lowest(P<0.05).At the third day,turbidity showed no difference between DATS group and negative control group(P=0.454).At the 7th day,between calcium hydroxide and the negative control group,P<0.05.Conclusion:DATS may suppress E.faecalis in the biofilm in vitro.
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Cardiovascular diseases(CVD) with high morbidity and mortality pose severe threats to human life. Allicin, a main active ingredient of garlic, possesses multiple pharmaceutical activities. It not only exerts cardioprotective effects but also prevents the risk factors for CVD. Allicin exerts cardioprotective effects via a variety of mechanisms, including inhibiting oxidative stress, apoptosis, autophagy, and inflammatory responses, regulating lipid metabolism and gut microbiota, inducing hydrogen sulfide production, and dilating vessels. Despite the valuable cardioprotective effects, the instability of allicin has hindered the basic research and clinical application. This paper reviews the progress in the cardioprotective effects and mechanisms of allicin in the last decade and summarizes the methods to improve the stability of allicin. In addition, this review provides a reference for further research and development of allicin in cardiovascular protection.
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Abstract The objective of this study was to evaluate the properties of garlic as a growth promoter in larvae of angelfish Pterophyllum scalare and its benefits during transport of juveniles of this species. The experiment was conducted in recirculation aquaculture system, consisting of 20 tanks of 40 L equipped with independent water input and output. We used 1,400 larvae distributed among five treatments, with four repetitions, which totaled 20 experimental units. The treatments were 0, 50, 100, 200, and 400 mg of garlic extract per kilo of feed. The results showed that the inclusion of levels of garlic extract in the feed did not significantly effect the fish's development or their transport. Neither did the inclusion of levels of garlic extract affect the survival of the larvae during the trial period. New research with extracts of higher dosages should be performed to elucidate the effect of garlic extract as a growth promoter.
Resumo O estudo teve como objetivo avaliar as propriedades do alho como promotor de crescimento em larvas de acará bandeira Pterophyllum scalare e seus benefícios no transporte de juvenis da mesma espécie. O experimento foi conduzido em sistema de recirculação composto por 20 aquários de 40 L de volume útil, dotado de entrada e saída de água independentes e teve a duração de 40 dias. O delineamento experimental adotado foi o inteiramente casualizado (DIC), com cinco tratamentos e quatro repetições, totalizando 20 unidades experimentais. Foram utilizadas 1.400 larvas de acará bandeira (Pterophyllum scalare) divididas entre os tratamentos. Os tratamentos consistiram em: 0, 50, 100, 200 e 400 mg de extrato de alho por quilo de ração. Os resultados mostraram que não houve efeito significativo dos níveis de inclusão do extrato do alho sobre os índices zootécnicos avaliados e posteriormente no transporte dos juvenis. Também não foi observado influência na sobrevivência das larvas durante o período experimental. Novas pesquisas com dosagens maiores de extratos devem ser realizados para melhor elucidação do efeito do extrato de alho como promotor de crescimento.
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Animais , Ciclídeos , Alho , Extratos Vegetais/farmacologia , Aquicultura , LarvaRESUMO
The objective of this study was to evaluate the properties of garlic as a growth promoter in larvae of angelfish Pterophyllum scalare and its benefits during transport of juveniles of this species. The experiment was conducted in recirculation aquaculture system, consisting of 20 tanks of 40 L equipped with independent water input and output. We used 1,400 larvae distributed among five treatments, with four repetitions, which totaled 20 experimental units. The treatments were 0, 50, 100, 200, and 400 mg of garlic extract per kilo of feed. The results showed that the inclusion of levels of garlic extract in the feed did not significantly effect the fishs development or their transport. Neither did the inclusion of levels of garlic extract affect the survival of the larvae during the trial period. New research with extracts of higher dosages should be performed to elucidate the effect of garlic extract as a growth promoter.
O estudo teve como objetivo avaliar as propriedades do alho como promotor de crescimento em larvas de acará bandeira Pterophyllum scalare e seus benefícios no transporte de juvenis da mesma espécie. O experimento foi conduzido em sistema de recirculação composto por 20 aquários de 40 L de volume útil, dotado de entrada e saída de água independentes e teve a duração de 40 dias. O delineamento experimental adotado foi o inteiramente casualizado (DIC), com cinco tratamentos e quatro repetições, totalizando 20 unidades experimentais. Foram utilizadas 1.400 larvas de acará bandeira (Pterophyllum scalare) divididas entre os tratamentos. Os tratamentos consistiram em: 0, 50, 100, 200 e 400 mg de extrato de alho por quilo de ração. Os resultados mostraram que não houve efeito significativo dos níveis de inclusão do extrato do alho sobre os índices zootécnicos avaliados e posteriormente no transporte dos juvenis. Também não foi observado influência na sobrevivência das larvas durante o período experimental. Novas pesquisas com dosagens maiores de extratos devem ser realizados para melhor elucidação do efeito do extrato de alho como promotor de crescimento.
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Animais , Alho , Dieta , Perciformes/crescimento & desenvolvimentoRESUMO
Abstract The objective of this study was to evaluate the properties of garlic as a growth promoter in larvae of angelfish Pterophyllum scalare and its benefits during transport of juveniles of this species. The experiment was conducted in recirculation aquaculture system, consisting of 20 tanks of 40 L equipped with independent water input and output. We used 1,400 larvae distributed among five treatments, with four repetitions, which totaled 20 experimental units. The treatments were 0, 50, 100, 200, and 400 mg of garlic extract per kilo of feed. The results showed that the inclusion of levels of garlic extract in the feed did not significantly effect the fishs development or their transport. Neither did the inclusion of levels of garlic extract affect the survival of the larvae during the trial period. New research with extracts of higher dosages should be performed to elucidate the effect of garlic extract as a growth promoter.
Resumo O estudo teve como objetivo avaliar as propriedades do alho como promotor de crescimento em larvas de acará bandeira Pterophyllum scalare e seus benefícios no transporte de juvenis da mesma espécie. O experimento foi conduzido em sistema de recirculação composto por 20 aquários de 40 L de volume útil, dotado de entrada e saída de água independentes e teve a duração de 40 dias. O delineamento experimental adotado foi o inteiramente casualizado (DIC), com cinco tratamentos e quatro repetições, totalizando 20 unidades experimentais. Foram utilizadas 1.400 larvas de acará bandeira (Pterophyllum scalare) divididas entre os tratamentos. Os tratamentos consistiram em: 0, 50, 100, 200 e 400 mg de extrato de alho por quilo de ração. Os resultados mostraram que não houve efeito significativo dos níveis de inclusão do extrato do alho sobre os índices zootécnicos avaliados e posteriormente no transporte dos juvenis. Também não foi observado influência na sobrevivência das larvas durante o período experimental. Novas pesquisas com dosagens maiores de extratos devem ser realizados para melhor elucidação do efeito do extrato de alho como promotor de crescimento.
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Resumen Las enfermedades cardiovasculares (ECV) comprenden un grupo de enfermedades cuyo denominador común es la afectación de vasos sanguíneos, corazón y ritmo cardiaco. El tratamiento de las ECV representa costos muy altos para los sistemas de salud y está enfocado en el control de los factores de riesgo. A pesar de existir una gran variedad de fármacos para el tratamiento de las ECV, estas continúan siendo las principales causas de mortalidad, posiblemente debido a que su origen es multifactorial y por ello se requiere de más de un fármaco. En este contexto, la alicina, un compuesto derivado del ajo, ha mostrado regular la expresión de vías de señalización y factores de riesgo asociados a la progresión de las ECV. Por ello el objetivo del presente trabajo es revisar los mecanismos celulares y moleculares por medio de los cuales la alicina ejerce sus efectos terapéuticos y describir las evidencias científicas del porqué la alicina podría representar un potencial candidato para coadyuvar en el tratamiento de las ECV.
Abstract Cardiovascular diseases (CVD) include a group of diseases whose common denominator is the affection of the blood vessels, heart, and heart rate. The treatment of CVD represents high costs to the health systems and is focused on the control of risk factors. Despite the existence of a great variety of treatments of the CVD, these continue as the main cause of mortality mainly due to the multifactorial origin, and therefore more than one drug is required. In this context, allicin, a compound derived from garlic, has shown regulate the expression of signaling pathways and risk factors associated with the progression of CVD. Therefore, the objective of this work is to review the cellular and molecular mechanisms through which allicin exert its therapeutic effects and to describe the scientific evidences why allicin represents a potential candidate to assist in the treatment of CVD.
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@#A novel allicin pro-drug tablet containing antacid pellets was developed to realize pH-regulated allicin release and to guarantee allicin yield in stomach environment.Firstly, allicin precursor pellets containing antacid pellet were prepared and artificial gastric juice was used as the medium to determine the yield of the allicin.Then, the total lipid cholesterol (TC), triglyceride cholesterol (TG), high density lipoprotein (HDL-C) and low-density lipoprotein (LDL-C) were used as indicators to study the hypolipidemic effect of allicin precursor pellets in rats.The dissolution test showed that in artificial gastric juice, the yield of allicin-containing antacid pellets exceeded 90%.In pharmacodynamic studies, it was found that antacid pellets showed the expected hypolipidemic effect on hyperlipidemia rats compared without antacid pellets.There was a very significant difference in blood lipid levels between the two test groups (P < 0.05).The allicin pro-drug tablets containing antacid pellets can effectively lower blood lipids.
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Garlic or Lashun is the member Liliaceae family, is use as spice in food cooking as well as medicine to treat various ailments. Garlic is also acting as a flavoring agent for the cooking, and however it has also been used as a drug from very ancient and modern times in all over the world, it is used to inhibit and cure the vast range of ailments and disorders. Allicin found in the garlic is the chemically active substance of fresh garlic extract, possess the capacity of readily permeable through phospholipid membranes which contributes to its possible pharmacological activity and also contain sulfur compounds, which are believed to bring some of the health benefits. Currently, garlic is broadly used for different diseases related with the systemic circulation and heart, which includes atherosclerosis, HDL, LDL & heart attack, coronary heart disease, and hypertension. Garlic is also reported to treat the lung cancer, and various other cancers such as colon cancer& skin diseases too, it also has hypolipidemic, immunomodulator, aphrodisiac, & Antifungal actions. This article reviews the importance of garlic (Allium sativum), and, their active constituents to show whether or not can be further used as potential natural sources for the development of any novel drug formulations.
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Objective:To investigate the effect of allicin (ALL) on learning and memory ability of rats with vascular dementia (VD) and the possible mechanism. Method:The VD rats induced by modified bilateral common carotid artery occlusion (BCCAO) were randomly divided into the VD group, low- and high-dose ALL (ALL-L and ALL-H) groups, and the sham operation (S) group, with 15 rats in each group. In the ALL-L and ALL-H groups, ALL was injected into the femoral vein at 5 mg·kg<sup>-1</sup> and 20 mg·kg<sup>-1</sup>, respectively, while the same volume of normal saline was injected in the S and VD groups, once a day, for two successive weeks. Morris water maze (MWM) was used to test the learning and memory ability of rats. Hematoxylin and eosin (HE) staining was conducted to observe the pathological changes in hippocampal tissue, followed by the detection of inflammatory factors tumor necrosis factor-<italic>α</italic> (TNF-<italic>α</italic>), interleukin-6 (IL-6), and IL-1<italic>β</italic> as well as oxidative stress indexes malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) in rat hippocampus. The apoptosis of hippocampal cells was detected by TdT-mediated dUTP Nick end Labeling(TUNEL) assay. The expression levels of apoptosis and autophagy-related proteins cysteinyl aspartate-specific protease-3 (Caspase-3), B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), microtubule-associated protein light chain 3Ⅱ (LC3Ⅱ), LC3Ⅰ, and the mammalian homolog of yeast ATG6 (Beclin 1) in hippocampus were determined by Western blot. Result:The comparison with the VD group revealed that the learning and memory abilities of rats in the ALL-H and ALL-L groups were significantly improved (<italic>P</italic><0.05). The TNF-<italic>α</italic>, IL-6, IL-1<italic>β</italic>, and MDA levels in hippocampus were lowered (<italic>P</italic><0.05), whereas the SOD and GSH-Px activities were enhanced (<italic>P</italic><0.05). The apoptosis rates were declined (<italic>P</italic><0.05), with an even lower rate noticed in the ALL-H group (<italic>P</italic><0.05). The expression levels of Caspase-3, Bax, LC3Ⅱ/LC3Ⅰ ratio, and Beclin-1 in the ALL-H and ALL-L groups were significantly down-regulated in contrast to those in the VD group (<italic>P</italic><0.05), while that of Bcl-2 was up-regulated (<italic>P</italic><0.05). The ALL-H group exhibited better performances than the ALL-L group (<italic>P</italic><0.05). Conclusion:ALL could improve the learning and memory ability of VD rats to some extent, which may be attributed to its inhibition against inflammatory reaction, oxidative stress, and neuronal apoptosis and autophagy.
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Objective @#To investigate the effect and mechanism of allicin combined with 5-FU on proliferation inhibition and apoptosis of the mucoepidermoid carcinoma MEC-1 cell line in mucoepidermoid carcinoma in order to provide the corresponding basis for subsequent clinical drug application.@*Methods @# MEC-1 cells in the logarithmic growth phase were randomly divided into control groups and experimental groups. The control groups were PBS groups containing 0.1% DMSO, while the experimental groups were the allicin group, 5-FU group and combined drug group (the allicin combined with the 5-FU group). The proliferation inhibition rates of allicin, 5-FU and allicin combined with 5-FU in MEC-1 cells were detected by the CCK8 method at different concentrations (0, 25, 50, and 75 mg/L) for 24 h, and the IC50 value of allicin and 5-FU after 24 hours was calculated. The apoptotic rate of MEC-1 cells treated with allicin, 5-FU and allicin combined with 5-FU at different concentrations (0, 25, 50, and 75 mg/L) for 24 hours was measured by flow cytometry. The expression of Bax and Bcl-2 protein was determined by Western blot analysis of the IC50 concentration of allicin and 5-FU alone and in combination with MEC-1 cells for 24 hours. @*Results@#The growth inhibition rate and apoptosis rate of MEC-1 cells in the combined drug group were higher than those in the allicin group and the 5-FU alone group (P < 0.01). Allicin and 5-FU alone and in combination downregulated Bcl-2 protein and upregulated Bax protein expression, and the combined drug group had the largest ratio of Bax/Bcl-2 (P < 0.05). @*Conclusion @#Allicin and 5-FU both alone and in combination can inhibit the proliferation of and induce apoptosis in MEC-1 cells, and allicin can enhance the apoptosis of 5-FU in MEC-1 cells, which may be related to the apoptosis of the mitochondrial pathway.
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Ulcerative colitis (UC) is a chronic, nonspecific intestinal inflammatory disease. Immune imbalance and intestinal mucosal barrier damage are important factors in the development and persistence of UC. In recent years, authors both domestic and abroad found that allicin could alleviate the intestinal mucosal inflammation of UC. This article reviewed the research progress on allicin in treatment of UC.
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Objective: To investigate the effects of allicin (All) in combination with lycopene (Lyc) against target organ damage and oxidative stress induced by hypertension in spontaneously hypertensive rats (SHRs). Methods: SHRs aged 10 weeks were randomly divided into four groups (n=8 in each group): SHR, SHR+All, SHR+Lyc, and SHR+All+Lyc. SHR in SHR+All, SHR+Lyc and SHR+All+Lyc groups were intraperitoneally or intragastrically administered with 15 or 7.5 mg/(kg•d) of All and Lyc at the same time daily for 6 weeks. Male WKY rats were intraperitoneally injected with the same volume of normal saline and served as normotensive controls (WKY group). Six weeks later, histopathological changes of thoracic aorta and kindeys were evaluated by HE staining and according to Paller's method. The indexes of oxidation (MDA) and antioxidants (SOD, CAT and GSH) in serum or thoracic aorta were determined by enzyme-linked immunosorbent assay (ELISA). Production of intracellular superoxide anion was stained by dihydroethidium (DHE) staining and detected using laser scanning confocal microscope. Results: SHR+Lyc+All group significantly ameliorated arterial wall thickening and inflammatory injury of the kindey in SHR compared with SHR+All group and SHR+Lyc group. Administration of All combined with Lyc significantly increased activities of antioxidative enzymes (SOD and GSH) and the content of CAT in aortas when compared with those of SHR and SHR receiving All or Lyc administration alone (P<0.05). In addition, the combined treatment of All with Lyc also significantly reduced the contents of MDA and O2- of SHR compared to SHR (P<0.05). Conclusion: This combination of All and Lyc can reduce target organ damage and level of oxidative stress induced by hypertension, which may be attributed to their regulations on ROS level, antioxidant and anti-inflammatory abilities.
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Objective The metastasis mechanism of cholangiocarcinoma is complex, which may be related to epithelial-mesenchymal transition(EMT). This study focused on investigating the inhibition effects of allicin on TGF-β1 induced epithelium mesenchymal transition of human cholangiocarcinoma cells and its related mechanism, and providing theoretical basis for the application of allicin in the treatment of cholangiocarcinoma. Methods MTT assay were used to detect the inhibition effects of different concentrations of allicin on the human cholangiocarcinoma RBE cell proliferation, and the drug concentration of allicin was determined by IC50 of 24 h. The RBE cells were cultured and divided into control group, allicin group(130.7μmol/L), TGF-β1 group(10ng/mL) and allicin+ TGF-β1 group(130.7μmol/L+10ng/mL). Wound scratch and transwell invasion assay were performed to detect the migration and invasion ability of RBE cells after 24 hours. Western blots were applied to detect expression of EMT-related proteins (E-Cadherin, N-Cadherin, Vimentin, Snail) and NF-κB signaling pathways. Results The migration rates in allicin group and allicin+ TGF-β1 group were both decreased compared with that in the control group ( 9.25% ± 0.36% vs 28.19 %±0.66%, P<0.05) and TGF⁃β1 group(13.91%±0.75% vs 49.22%±0.27%, P<0.05). The invasion rates in allicin group and allicin+ TGF-β1 group were also decreased compared with that in the control group (6.59%±0.06% vs 33.48%±0.04%, P<0.05) and TGF⁃β1 group(9.4%± 0.05% vs 40.21%±0.12%, P<0.05). Compared with the control group, E-Cadherin expression was significantly increased, and N-Cadherin, Vimentin, Snail, NF-κB and p-NF-κB expression were significantly decreased in the allicin group (P<0.05). Compared with TGF-β1 group, E-Cadherin expression was significantly up-regulated, and N-Cadherin, Vimentin, Snail, NF-κB and p-NF-κB expression were significantly down-regulated in the allicin+ TGF-β1 group (P<0.05). Conclusion These results indicate that allicin can inhibit the EMT induced by TGF-β1 on the human cholangiocarcinoma cell by blocking NF-κB signaling pathway, which may have potential value to be the drug candidate for the treatment of human cholangiocarcinoma in future.
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Objective:To prepare self-emulsifying carrier system which was suitable for three Chinese medicinal ingredients(baicalein,berberine and allicin),and investigate transdermal absorption effect in vitro of these three self-emulsifying preparations. Method:The optimum formulation and dosage were screened by the saturated solubility method,pseudo-ternary phase diagram method and orthogonal experiment.Transdermal absorption test in vitro was carried out with excised rats skin and Franz diffusion cell.The cumulative penetration amounts of baicalein,berberine and allicin in the identical self-emulsifying system were determined by HPLC and compared with baicalein powder,berberine powder and allicin powder,respectively. Result:The optimum formulation was ethyl oleate-cremophor RH40-polyethylene glycol(PEG)400.The self-emulsifying preparation had a suitable particle size with a relatively regular spherical shape.At 10 h of transdermal absorption,the transdermal rates of baicalein,berberine and allicin in identical self-emulsifying system were 6.898 6,7.600 4,190.040 μg·cm-2·h-1,the cumulative penetration amounts of them were 71.38,85.54,1 795.16 μg·cm-2,respectively. Conclusion:The self-emulsifying carrier system is prepared successfully,which can be used by different kinds of Chinese medicinal ingredients,and the transdermal absorption effect in vitro of these self-emulsifying preparations is good,which can provide experimental basis for the preparation and transdermal absorption of self-emulsifying preparation of Chinese herbal compound.
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OBJECTIVE: To study the absorption characteristics of allicin in rat intestinal tract.METHODS: An HPLC method was established to determine the content of allicin in rat entericus, and Ussing chamber system was used to investigate the absorption characteristics of garlic spicy element of different concentrations in the duodenum, jejunum, ileum and colon of rats. RESULTS: An HPLC method was established to determine the content of allicin in rat intestinal liquid.The absorption rate constants of allicin in various intestinal segments of rats in the experimental concentration range increased with the increase of allicin concentration.Allicin showed linear absorption in different intestinal segments of rats, with regression correlation coefficients of greater than 0.96, indicating zero order absorption. CONCLUSION: Allicin has different absorption characteristics in different intestinal segments of rats and can be absorbed as prototype.
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Objective To investigate the protective effect and mechanism of garlicin on oxidative stress injury of human melanocytes.Methods There were blank group,control group,hydrogen peroxide group,garlicin group,experiment 1,2 and 3 groups.No cells in the blank group were only added with complete culture medium.The control group was added with complete medium;0.4 mmol/L hydrogen peroxide complete medium was added to the hydrogen peroxide group.Garlicin group was added with freshly prepared garlicin complete culture medium with concentration of 40 μmol/L;Experiment groups 1,2,and 3 were treated with different concentrations of garlicin (80,40,and 20 μmol/L garlicin complete medium,respectively) to interfere with melanocytes treated with 0.4 mmol/ L hydrogen peroxide during logarithmic growth period.After 24 h of drug intervention,the cell morphology was observed under an inverted microscope.The protective effect of garlicin on melanocytes damaged by oxidative stress was measured by MTT colorimetric method.Results Different concentrations of garlicin had different protective effects on melanocytes induced by hydrogen peroxide.It could be concluded that the activity of melanocytes in hydrogen peroxide group decreased significantly (43.610 ± 3.872)% (P<0.05),but there was no statistical difference between the two groups (P=0.345).The activity of melanocytes in experimental group 1 was significantly decreased (58.223 ± 2.806) % but higher than that in experimental groups 2 and 3 (P<0.05).Conclusions Allicin inhibits the production of intracellular ROS in human melanocytes induced by H2 O2,regulates oxidative stress in human melanocytes,and counteracts H2O2-induced apoptosis.Therefore,allicin may be a protective factor in mediating oxidative stress in the body.
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Objective To investigate the inhibitory effect of Allicin on the apoptosis of hippocampal neurons induced by lead in rats.Methods Sixty male Sprague-Dawley rats aged 3 weeks were randomly divided into 6 groups,10 rats in each group,which were low dose group(A-L),medium-dose group(A-M) and high dose (A-H) Allicin group and lead exposure group (Pb group),dimercaptosuccinic acid (DMSA) group and blank control group.The blank control group animals were treated with ultrapure water,and the other 5 groups received 1.0 g/L lead acetate aqueous solution instead of ultrapure water after 20 days and they were treated them with compounds by oral gavage.The doses of Allicin in group A-L,A-M group and A-H group were 2.7 mg/kg and 5.4 mg/kg,and 10.8 mg/kg,respectively.The DMSA dose was 10.8 mg/kg,and the Pb group was given 9 g/L saline.After the model was established,the rats were sacrificed to collect whole blood and hippocampus.Blood lead and tissue lead concentrations were measured,and the level of apoptosis in hippocampus was observed by TUNEL staining.The levels of cysteine-containing aspartate-specific proteases (caspase)-3,caspase-9,poly adenosine diphosphate-ribose polymerase (PARP) mRNA and caspase-3,caspase-9,PARP activated protein and cytochromes C distribution in the hippocampus cells were detected by using real-time quantitative PCR (qPCR),Western blot,and immunofluorescence staining.Results (1) Lead levels in the blood lead and hippocampus of rats in A-L group,A-M group and A-H group [(190.54±11.33) μg/L,(0.28 ±0.03) μg/L;(159.55 ±16.94) μg/L,(0.22 ±0.06) μg/L;(l16.62 ±8.85) μg/L,(0.19 ±0.01) μg/L] were lower than those in Pb group [(271.34 ±21.23) μg/L,(0.31 ±0.04) μg/L],and there were significant differences (all P < 0.05).The blood lead and hippocampal lead levels in the DMSA group [(50.12 ± 7.44) μg/L,(0.15 ± 0.03) μg/L] were lower than those in the A-L group,A-M group and A-H group.(2) The results of TUNEL staining showed that the apoptosis levels of hippocampus in A-L group,A-M group and A-H group were lower than that in Pb group [(2.81 ±0.17)%,(2.08 ±0.28)%,(1.33 ±0.08)% vs.(4.23 ±0.17)%],and there were significant differences (all P < 0.05);the apoptosis level of hippocampus in the DMSA group [(2.63 ± 0.32) %] was higher than that in the A-M group and the A-H group,which was lower than that in the Pb group.(3) qPCR results showed that the levels of caspase-3,caspase-9 and PARP mRNA in A-H group were down-regulated compared with Pb group (1.07 ± 0.05,1.02 ± 0.02,1.11 ± 0.02 vs.1.34 ± 0.02,1.26 ±0.05,1.93 ± 0.07).The differences were statistically significant (P < 0.05).The expression levels of caspase-3 and PARP mRNA in A-L group and A-M group were down-regulated (1.21 ± 0.05,1.43 ± 0.12,1.16 ± 0.02,1.20 ± 0.06 vs.1.34 ± 0.02,1.93 ± 0.07),and there were significant differences (all P < 0.05),and there was no significant change in caspase-9 mRNA;the mRNA levels of caspase-3,caspase-9 and PARP in A-H group (1.07 ± 0.05,1.02 ± 0.02,1.11 ± 0.02) were lower than those in DMSA group (1.14 ± 0.02,1.15 ± 0.08,1.32 ±0.05).(4) Western blot results:compared with Pb group,the expression levels of activated caspase-3,caspase-9 and PARP protein in A-H group were down-regulated (A-H group:0.44 ± 0.15,0.58 ± 0.25 and 0.31 ±0.19,0.23 ±0.07 vs.Pb group:0.69 ±0.13,0.72 ±0.22 and 0.55 ±0.21,0.43 ±0.10),the expression of activated caspase-9 protein in A-M group was lower than that in Pb group (A-M group:0.59 ±0.18 vs.Pb group:0.72 ± 0.22),and there were significant differences (all P < 0.05);the expression of activated caspase-3 and RARP protein in A-H group was lower than that in DMSA group.(5) Fluorescence staining showed that the expression of cytochrome C in cytoplasm of A-L group,A-M group and A-H group were significantly lower than that of Pb group and DMSA group.Conclusion Allicin can inhibit the apoptosis of hippocampus cells in rats with lead poisoning through mitochondrial pathway.The effect of Allicin on apoptosis inhibition may be better than DMSA.
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Allicin is one of the main bioactive substances in garlic, with antibacterial, hypolipidemic and other pharmacological effects. In this study, apoptosis-related indicators were detected to explore the molecular mechanism of allicin on KG-1 cell proliferation inhibition. The apoptosis rate of KG-1 cells induced by allicin was detected by flow cytometry. The effect of allicin on the expressions of Bax, Bcl-2, survivin and ERK mRNA in KG-1 cells was detected by RT-qPCR. Western blot was used to detect the expressions of caspase 3, cleaved caspase 3, ERK1/2, p-ERK1/2 and survivin protein in KG-1 cells. According to the findings, compared with the control group, allicin could significantly inhibit the proliferation activity of KG-1 cells in a concentration-dependent and time-dependent manner. Flow cytometry showed that allicin could induce the apoptosis of KG-1 cells, which was mainly late apoptosis. The results of RT-qPCR showed that the expressions of Bax mRNA, Bcl-2, survivin and ERK mRNA in KG-1 cells increased after treatment with allicin. The results of Western-blot showed that after KG-1 cells were treated with allicin, the expressions of caspase 3 and its active form cleaved caspase 3 increased, the expressions of survivin, ERK1/2 and its active form p-ERK1/2 were decreased, of which p-ERK1/2 was down-regulated in a dose-dependent manner. The above results suggest that allicin inhibited the proliferation of KG-1 cells primarily by inducing late apoptosis; the execution of apoptosis involved cleaved caspase 3; the induction of apoptosis involved the protein expression, the decrease of ERK1/2andexpression of survivin and the dose-dependent decrease of p-ERK1/2; the mRNA expression involved the increase of Bax, and the down-regulation of survivin, Bcl-2 and ERK1/2.
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Objective To investigate the effect of allicin on the proliferation,apoptosis and invasion of human multiple myeloma (MM) cell line RPMI8226 in vitro,and to explore its mechanism.Methods The human MM cell line RPMI8226 cells were treated with different concentrations of allicin as 0,5,25,125 μmol/L,wherein the control group was 0 μmol/L allicin treatment group.The proliferation of cells was calculated by cell counting kit-8 (CCK-8).The apoptosis and invasion ability of tumor cells were determined with flow cytometry and Transwell method respectively.The expressions of transforming growth factor-α (TGF-α),matrix metalloproteinase (MMP)-2,MMP-9 and tissue inhibitor of metalloproteinase-2 (TIMP-2)protein were detected by Western blotting.Results The proliferation rates of RPMI8226 cells in groups treated with 0,5,25 and 125 μmol/L allicin were 1.00% ± 0.02%,0.98% ± 0.04%,0.73% ± 0.22% and 0.44% ± 0.15% respectively,with a significant difference (F =329.2,P < 0.001).The proliferation rates of RPMI8226 cells in the 25 and 125 μmol/L allicin treatment groups were significantly inhibited compared with control group (P <0.001;P <0.001);but there was no significant difference between 5 μmol/L treatment group and control group (P=0.395).The apoptosis rates of RPMI8226 cells treated with 0,5,25 and 125 μmol/L allicin were 3.05% ±0.53%,4.06% ±0.29%,12.17% ± 1.08% and 12.81% ± 1.78% respectively,with a significant difference (F =531.0,P <0.001).The apoptotic rates of RPMI8226 cells in the 25 and 125 μmol/L allicin treatment groups were significantly increased compared with control group (P =0.013;P =0.012);and there was no significant difference between 5 μmol/L treatment group and control group (P =0.211).The numbers of invasive cells in the 0,5,25 and 125 μmol/L allicin treatment groups were 112.5 ± 1.9,104.8 ±4.0,76.9 ± 2.6 and 52.5 ± 3.7 respectively,with a significant difference (F =734.9,P < 0.001).The abilities of cells invasiveness in 25 and 125 μmol/L allicin treatment groups were significantly decreased compared with control group (P < 0.001;P < 0.001),but 5 μmol/L allicin treatment group had no significant difference compared with the control group (P =0.160).Western blotting results showed that the expression levels of TGF-α,MMP-2,MMP-9 and TIMP-2 proteins were significantly different between groups treated with different concentrations of allicin (F =227.1,P<0.001;F=348.5,P<0.001;F=359.7,P<0.001;F=158.0,P <0.001).The protein expression levels of TGF-eα,MMP-2 and MMP-9 were significantly decreased in the 25 and 125 μmol/L treatment groups compared with the control group (all P < 0.001),and the expression of TIMP-2 protein was significantly increased compared with the control group (all P < 0.001),but there were no significant diffe-rences between the 5 μmol/L treatment group and the control group (P =0.349;P =0.744;P =0.613;P =0.567).Conclusion Allicin can significantly inhibit cell proliferation,invasive ability and induce apoptosis of human MM cell line RPMI8226 in vitro,and the mechanism may be related to inhibiting the expressions of TGF-α,MMP-2,MMP-9 and promoting the expression of TIMP-2 at the same time.
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Objective To observe the effect of Allicin on renal tissue fibrosis in chronic renal failure (CRF) and to study the protective mechanism of Allicin on CRF rats. Methods The CRF rat models were established by intragastric administration of adenine solution (250 mg/kg) for 21 days. After successful modeling, 80 model rats were randomly divided into the normal control group, the model group and the allicin low, medium and high dose groups using a random number table method (n=20). Allicin low-, medium-, and high-dose groups were intragastrically administered with 2.5, 5, and 10 mg/ml of allicin respectively. The model group and normal control group were given an equal volume of normal saline, and were administered at 2 ml/kg body weight of the rats, with once a day continuous administration for 28 d. Two hours after the last dose, the kidney mass was measured and the kidney index was calculated. The pathological changes of renal tissue was observed by HE staining; the serum BUN, SCr, uric acid, and 24-hour urinary protein levels were measured by a biochemical analyzer; the levels of CRP, IL-6 and TNF-α in serum and the levels of ollagen type Ⅳ(CⅣ), type Ⅲ procollagen (PCⅢ), laminin (LN), Fibronectin (FN) in plasma were by ELISA; the expression of collagen type Ⅰ (Col Ⅰ), plasminogen activator inhibitor-1 (PAI-1) and MMP-1 were observed by mmunohistochemical staining. Results Compared with model group, the bodyweight of rats in allicin medium, high dose groups were increased and kidney index decreased (P<0.05 or P<0.01), kidney histopathology scores decreased (P<0.01). Compared with model group, the level of BUN (14.51 ± 2.76 mmol/L, 11.48 ± 2.43 mmol/L vs. 24.07 ± 3.82 mmol/L), SCr (116.28 ± 27.35 μmol/L, 106.57 ± 24.18 μmol/L vs. 134.89 ± 35.02 μmol/L), Uric acid (83.34 ± 16.42 mmol/L, 77.86 ± 13.97 mmol/L vs. 114.76 ± 16.53 mmol/L), 24 h urinary protein (152.79 ± 48.43 mg, 137.03 ± 42.61 mg vs. 177.94 ± 96.47 mg), CRP (8.79 ± 1.84 mg/L, 7.51 ± 1.69 mg/L vs. 11.64 ± 1.95 mg/L), TNF-α (184.37 ± 24.15 ng/L, 126.82 ± 12.96 ng/L vs. 255.87 ± 31.93 ng/L) in the allicin medium and high dose groups were significantly decreased (P<0.05 or P<0.01); the levels of C-Ⅳ (40.26 ± 7.12 ng/ml, 23.79 ± 4.25 ng/ml vs. 67.53 ± 8.39 ng/ml), PC-Ⅲ (32.03 ± 5.89 ng/ml, 24.31 ± 5.84 ng/ml vs. 54.20 ± 7.08 ng/ml), LN (99.05 ± 38.17 ng/ml, 83.42 ± 28.83 ng/ml vs. 117.83 ± 35.76 ng/ml) in plasma in the allicin medium and high dose groups were significantly decreased and the level of FN (98.58 ± 21.43 mg/L, 125.96 ± 25.12 mg/L vs. 66.72 ± 13.09 mg/L) in plasma in the allicin medium and high dose groups were significantly increased (P<0.05 or P<0.01); the expression of Col Ⅰ (0.17 ± 0.03, 0.09 ± 0.03 vs. 0.27 ± 0.05), PAI-1 (0.20 ± 0.05, 0.16 ± 0.04 vs. 0.31 ± 0.08) were down-regulated and the expression of MMP-1 (0.10 ± 0.03, 0.22 ± 0.05 vs. 0.04 ± 0.02) were up-regulated (P<0.05 or P<0.01). Conclusion Allicin has protective effects on CRF rats by inhibiting the renal tissue fibrosis and alleviating inflammation.