Effect of garlic and garlic preparations on physiological and psychological stress in mice

The effect of several kinds of garlic preparations such as raw garlic juice (RGJ), heated garlic juice (HGJ), processed garlic powder (PGP) and aged garlic extract (AGE) on both physiological and psychological stress were investigated using four stress models in mice: forced swimming test, mechanical treadmill running, immobilization stress test, and a cooling rewarming test. RGJ was shown to be effective only at a low dose in the forced swimming test, whereas the effect was reduced at a high dosage. HGJ and PGP demonstrated no antistress effects. In contrast, AGE was shown to be effective in all of the stress tests. © 1997 by John Wiley & Sons, Ltd.     

Identification of the protein components displaying immunomodulatory activity in aged garlic extract

Ethnopharmacological relevance

Traditionally, garlic (Allium sativum L.; Alliaceae) has been known to boost the immune system. Aged garlic has more potent immunomodulatory effects than raw garlic. These effects have been attributed to the transformed organosulfur compounds; the identity of the immunomodulatory proteins in aged garlic extract (AGE) is not known.

Aim of the study

The major aims are to examine the changes occurring in the protein fraction during ageing of garlic and to identify the immunomodulatory proteins.

Materials and methods

Changes occurring in garlic during ageing have been examined by protein quantitation and gel electrophoresis. Purification and identification of the immunomodulatory proteins have been achieved by Q-Sepharose chromatography and mitogenic activity.

Results

Only two major proteins (12–14 kDa range by SDS-PAGE) are observed in AGE. The purified protein components QA-1, QA-2, and QA-3 display immunomodulatory and mannose-binding activity; QA-2 shows the highest mitogenic activity. The identity of QA-2 and QA-1 proteins with the garlic lectins ASA I and ASA II, respectively, has been confirmed by hemagglutination analysis. QA-3 exhibits mitogenic activity, but no hemagglutination activity.

Conclusions

The immunomodulatory activity of AGE is also contributed by immunomodulatory proteins. The major immunomodulatory proteins have been identified as the well-known garlic lectins.     

Hepatoprotective effects of propolis extract on paracetamol-induced liver damage in mice

The effects of propolis extract were studied in a model of acute hepatotoxicity induced by a high oral dose (600 mg/kg) of paracetamol in mice. Propolis at doses of 25, 50 and 100 mg/kg i.p. decreased significantly the activity of alanine aminotransierase in serum, which was increased by paracetamol alone and increased the concentration of reduced glutathione in mouse liver, which is depleted by paracetamol. Propolis extract also reduced liver damage induced by paracetamol in mice, which was observed by optical and electron microscopy. The hepatoprotective effects of propolis were produced when administered 30 min before paracetamol or 2 h after it. It is concluded that propolis exerts some effects which resemble those of N-acetylcysteine, the well-known antidote of paracetamol.     

黑蒜水提液对小鼠肝癌H22移植性实体瘤的生长干预

目的:探讨黑蒜水提液对人肝癌SMMC 7721细胞及小鼠肝癌Hep.A22移植性实体瘤生长的作用。方法:体外培养SMMC 7721细胞,观察黑蒜水提液干预后对细胞增殖的影响;将昆明小鼠移植H22肝癌细胞后,分阴性对照组,环磷酰胺组,黑蒜水提液低、中、高剂量组,联合加热治疗组6个组,计算黑蒜水提液对肝癌实体瘤的抑瘤率、胸腺指数和脾指数;对小鼠血清细胞因子IL-2含量测定。结果:黑蒜水提液对肝癌肿瘤均有明显的抑制生长作用,抑瘤有效率可大于40%。结论:通过确定黑蒜水提液作为纯中药抗肿瘤药物的抑瘤作用,为临床应用提供实验依据。     

Effect of aged garlic extract against methotrexate-induced damage to the small intestine in rats

Methotrexate (MTX) chemotherapy is often accompanied by side effects such as gastrointestinal ulceration and diarrhea. The aim of this study was to examine histologically whether an aged garlic extract (AGE) had a protective effect on the small intestine of rats with MTX-induced damage. Forty male Wistar albino rats were randomized into experimental and control groups and divided into four groups of ten animals. To the first group, MTX was applied as a single dose (20 mg/kg) intraperitoneally. To the second group, in addition to MTX application, AGE (250 mg/kg) was administered orally every day at the same time by intragastric intubation until the rats were killed. To the third group, AGE only was given. The fourth group was the control. All animals were killed 4 days after the intraperitoneal injection of MTX for histopathologic analysis and tissue MDA levels. Before killing, intracardiac blood was obtained from each animal to perform biochemical analysis (plasma lactate level). MTX was found to lead to damage in the jejunal tissues and to increase the MDA and lactate levels in the plasma. Administration of the AGE decreased the severity of jejunal damage, but increased MDA and lactate levels caused by MTX treatment on the other hand. These results suggest that AGE may protect the small intestine of rats from MTX-induced damage. Thus this study substantiated the thought that the protective effect of AGE is derived from the manner in which it interacts with crypt cells.     

Aged garlic extract restores nitric oxide bioavailability in cultured human endothelial cells even under conditions of homocysteine elevation

Ethnopharmacological relevance

Supplementation with aged garlic extract (AGE) has been shown to restore impaired endothelium-dependent vasodilator response in subjects with acutely elevated plasma homocysteine (Hcy) levels after an oral methionine load and in patients with chronic coronary artery disease. Moreover, AGE has been shown to inhibit the progression of coronary calcifications in patients with coronary artery disease. The molecular mechanisms, by which AGE preserves endothelial function is unknown. Our objective was to explore whether AGE preserves endothelial nitric oxide (NO) output even under conditions of elevated Hcy levels by preventing oxidative inactivation of the NO synthase cofactor tetrahydrobiopterin.

Material and methods

Endothelial (EA.hy 926) cells were incubated with hypoxanthine, aminopterin, thymidine and methionine (HAT/MET) to increase cellular Hcy levels, and with and without AGE. Agonist stimulated NO output was measured using the fluorescent probe DAF-2, and cellular thiol levels (Hcy, cysteine, reduced and oxidized glutathione) and cellular tetrahydrobiopterin levels were measured by high performance liquid chromatography.

Results

HAT/MET incubation resulted in significantly increased cellular Hcy levels, unaffected by coincubation with AGE. Elevated Hcy went along with significantly decreased NO output (to 34.4±4.4% of control) and levels of tetrahydrobiopterin (from 4.67±2.17 to 2.17±0.97 pmol/mg). Incubation with AGE (5 mg/mL) in HAT/MET-treated cells prevented the declines in NO output and tetrahydrobiopterin levels. AGE increased cellular levels of cysteine and total glutathione, and prevented glutathione and tetrahydrobiopterin oxidation induced by elevated Hcy.

Conclusion

Incubation with AGE preserved normal NO output from endothelial cells even under conditions of elevated Hcy levels by increasing cellular thiol antioxidant and prevention of tetrahydrobiopterin oxidation. This suggests that AGE might be useful in the prevention of endothelial dysfunction.     

Use of crude extract of garlic (Allium sativum L.) in reducing cytotoxic effects of arsenic in mouse bone marrow

A relatively high concentration of crude extract of garlic, (Allium sativum L.)--single clove variety, was found to reduce the cytotoxic effects of three doses of sodium arsenite, corresponding to 1/10, 1/30 and 1/50 of the LD₅₀ in laboratory bred male Swiss albino mice. The animals were given a single oral dose of the chemical, together with the extract, and effects were observed after 24 h in bone marrow cells following the colchicine–fixation–airdry–Giemsa schedule. The endpoints scored were chromosomal aberrations and damaged cells.     

Aged garlic extract inhibits homocysteine-induced scavenger receptor CD36 expression and oxidized low-density lipoprotein cholesterol uptake in human macrophages in vitro

Aim of the study

Expression of CD36 scavenger receptors on macrophages is involved in oxidized low-density lipoprotein (OxLDL) uptake and foam cell formation during atherosclerotic lesion development. It has been shown previously in vitro and in vivo that the atherosclerotic risk factor homocysteine (Hcy) stimulates macrophage CD36 expression and OxLDL uptake. We now examined the effects of aged garlic extract (AGE), a garlic preparation enriched in water-soluble organic sulfur-containing compounds, on Hcy-induced CD36 expression and foam cell formation in human monocytes/macrophages.

Results

Incubation with Hcy (200 μM for 72 h) in THP-1-derived macrophages and primary human macrophages caused a 37.8 ± 5.2% and 60.7 ± 4.2% increase in CD36 expression compared to control, respectively. Coincubation with AGE (5 mg/ml) significantly suppressed CD36 expression in THP-1 derived macrophages by 48.6 ± 9.0% compared to Hcy-incubated cells only. AGE (1-5 mg/ml) dose dependently inhibited Hcy-induced CD36 expression in primary human macrophages, and decreased binding of nuclear proteins to a PPARγ response element. Preincubation with AGE significantly inhibited DiI-labeled OxLDL uptake.

Conclusions

These data indicate that AGE inhibits CD36 expression and OxLDL uptake in human macrophages by modulating the PPARγ pathway, and suggest that the extract could be useful for the prevention of atherosclerotic lesions.     

Activation of immunoresponder cells by the protein fraction from aged garlic extract

F-4, a protein fraction isolated from aged garlic extract strongly stimulates mice peritoneal macrophages in terms of glucose utilization and exhibits cytostatic activity in vitro. F-4 also has a mitogenic activity on spleen cells. In vivo, F-4 induced the stimulation of carbon clearance activity in mice. No obvious activities were observed in the low molecular fraction of aged garlic in these tests, but weak effects were found in the fructan fraction (F-3).     

Amelioration of early cognitive deficits by aged garlic extract in Alzheimer's transgenic mice

Subtle accumulation of beta-amyloid peptide (Abeta) oligomers of Abeta42 species in particular, is known to correlate with cognitive deficits independent of Abeta plaque deposition in the brain. Majority of the research showing behavioral improvement after cerebral Abeta reduction has been reported when the animals carried fewer/abundant amyloid plaques in the brain. Very few studies have addressed whether or not behavioral deficits exist even at the pre-plaque stage or in the absence of plaques that would parallel the mild cognitive impairment (MCI) stage of Alzheimer's disease (AD). Current study was undertaken to determine whether there exists any cognitive impairment during the pre-plaque stage which may parallel the MCI stage of AD, and to confirm whether the observed behavioral deficits correlate with Abeta42 predominance. In addition, the study determined whether anti-amyloidogenic effects of dietary aged garlic extract would prevent progressive behavioral impairment. For this purpose we used Tg2576 model showing slow plaque development with a predominance of Abeta40, and the TgCRND8 model showing accelerated plaque development with a predominance of Abeta42. The results show that at 2 months of age Tg2576 mice did not exhibit behavioral impairment in any of the tasks studied. While 2-month-old TgCRND8 mice displayed only a subtle behavioral deficit that matched the behavioral deficits observed in 7-month-old Tg2576 mice which may correlate with the MCI stage of AD. TgCRND8 mice at 7 months of age exhibited advanced deterioration in all behavioral tasks studied, suggesting that accelerated Abeta accumulation and the predominance of Abeta42 species may account for the pronounced cognitive deficits observed in TgCRND8. Feeding of aged garlic extract prevented deterioration of hippocampal based memory tasks in these mice, suggesting that aged garlic extract has a potential for preventing AD progression.     

Prevention of bone loss by oil extract of garlic (Allium sativum Linn.) in an ovariectomized rat model of osteoporosis

The effects of oil extract of garlic (Allium sativum Linn.) on different primary and secondary osteoporotic marker changes were tested in an ovariectomized rat model of osteoporosis. Experiments were performed on three different rat models: sham-operated control, ovariectomized and ovariectomized supplemented with garlic oil. In ovariectomized group, there has been a significant increase in different relative organ weights compared to sham-operated control, while the uterine weight was found to be decreased. Supplementation with oil extract of garlic could effectively reverse these changes. Also low bone densities that developed in the ovariectomized group were significantly recovered in the garlic oil supplemented group. In our study, the development of high rate of bone turnover and osteoporosis in the ovariectomized animals were confirmed by significant alteration of serum alkaline phosphatase activity, serum tartrate resistant acid phosphatase activity, urinary excretion of calcium, phosphate, hydroxyproline and urinary calcium to creatinine ratio, when compared with the sham-operated control group. Garlic oil extract supplementation, apart from its unique influence in lowering blood cholesterol, could also prevent ovariectomy-induced rise in all the above-mentioned marker changes. The results of this study emphasize that oil extract of garlic possibly has a positive role in suppressing ovariectomy-induced bone resorption.     

Aged garlic extract induces proliferation and ameliorates gentamicin-induced toxicity in LLC-PK1 cells

Gentamicin (GM)-induced nephrotoxicity limits the use of this antibiotic. It has been shown that aged garlic extract (AGE) and S-allylcysteine (SAC), the most abundant organosulfur compound in AGE, ameliorate GM-induced nephrotoxicity in rats. The present communication evaluated the effect of AGE and SAC on proliferation and on GM-induced toxicity and genotoxicity of porcine kidney epithelial cell line (LLC-PK1 cells). The cells were preincubated with different concentrations of AGE or SAC for 12 h before incubation with 8 mm GM for an additional 72 h. At the end of this time, cell viability, genotoxicity and proliferation were evaluated. AGE stimulated cell proliferation and protected LLC-PK1 cells from GM-mediated toxicity and genotoxicity. SAC partially prevented only GM-induced genotoxicity. These results suggest that the stimulation of cell proliferation could possibly be one of the mechanisms involved in the in vitro protective effect of AGE in GM-induced toxicity of LLC-PK1 cells.     

Aqueous garlic extract attenuates hepatitis and oxidative stress induced by galactosamine/lipoploysaccharide in rats

Injection of D-galactosamine and lipopolysaccharide (DGaIN/LPS) is useful as an experimental model of acute hepatic damage. Juvenile rats were used for investigation. The hepatoprotective activity of aqueous garlic (Allium sativum) extract (AGE) at a dose of 300 mg/kg body weight for 14 days, intraperitoneal (i.p.) prior to the induction of DGalN/LPS, was investigated against DGalN/LPS-induced hepatitis in rats. DGalN/LPS (300 mg/kg body weight/30 microg/kg body weight, i.p.), induced hepatic damage that was manifested by a significant increase in the activities of marker enzymes [alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), lactate dehydrogenase (LDH) and gamma glutamyl transferase (gamma GT)], bilirubin, lipid peroxides (LPO), tumor necrosis factor (TNF-alpha) and myeloperoxidase (MPO) activity level in serum. Also, the lipid profile in serum and liver homogenate including total cholesterol, triglycerides, free fatty acids and phospholipids were significantly deteriorated. The antioxidant enzyme activities (superoxide dismutase, SOD; reduced glutathione, GSH; catalase, CAT and glutathione peroxidase, GPX) in liver homogenate were significantly decreased in the DGalN/LPS. Pretreatment of rats with AGE reversed these altered parameters near to normal control values. Results of this study revealed that AGE could afford a significant protection in the alleviation of DGalN/LPS-induced hepatic damage.     

Role of oil extract of garlic (Allium sativum Linn.) on intestinal transference of calcium and its possible correlation with preservation of skeletal health in an ovariectomized rat model of osteoporosis

The present study was undertaken to examine the effects of an oil extract of garlic on the in vivo intestinal transference of calcium, and also to verify its role in maintaining the bone mineral content and bone tensile strength in an ovariectomized rat model of osteoporosis. The results suggest that, in this experimental model, oil extract of garlic promotes intestinal transference of calcium by modulating the activities of both intestinal alkaline phosphatase and Ca(2+) activated ATPase. Also the observed low bone mineral content and low bone tensile strength in these rats were significantly restored by garlic oil supplementation. Further, garlic oil supplementation was able to revive partially the bilateral ovariectomy-induced decrease in the serum estrogen titer. The serum parathyroid hormone level, however, was found unaltered in these rats. The garlic oil supplemented partial recovery in serum estrogen titer in bilaterally ovariectomized rat was found to be persistently associated with enhanced calcium transference and better preservation of bone mineral content. The results of this study propose that the phytoestrogenic efficacy of an oil extract of garlic prevents ovarian hormone deficiency induced bone mineral loss possibly by promoting intestinal transference of calcium through the partial revival of the serum estrogen titer.     

重楼对小鼠急性肝损伤保护作用的研究

目的:探讨重楼醇提物和水提物对四氯化碳(CCl4)所致小鼠急性肝损伤的保护作用。方法:小鼠随机分为9组,分别每天给予不同剂量的药物:重楼醇提物高、中、低剂量组(3.9、2.6、1.3 g/kg)、水提组的高、中、低剂量组(7.2、4.8、2.4g/kg)和联苯双酯(0.15 g/kg),正常组和模型组给予生理盐水。7d后,除正常对照组外各组小鼠腹腔注射0.1%CCl4制备急性肝损伤模型,16h后取血检测血清ALT、AST活性;测定肝组织匀浆中MDA含量和SOD、GSH的活性及观察肝脏病理学变化。结果:醇提组高剂量(3.9g/kg)和水提物高、中、低剂量(7.2、4.8、2.4g/kg)组均能降低CCl4所致急性肝损伤小鼠血清ALT、AST活性(P<0.01,P<0.05),降低肝脏MDA的含量、增强SOD和GSH活力(P<0.01,P<0.05),并能明显改善肝组织的病理学损伤。结论:重楼水提物对CCl4所致小鼠急性肝损伤具有较好保护作用。     

Inhibitory actions of Ganoderma lucidum spore extract on delayed-type hypersensitivity in mice

The effect of Ganoderma lucidum spore extract (GLSE) on murine delayed-type hypersensitivity (DTH) induced by sheep red blood cells (SRBC), 2,4-dinitrochlorobenzene, and allotypic splenocytes was studied. It was found to inhibit the DTH reaction significantly. It is concluded that GLSE may inhibit cell-mediated immunity in mice.     

Glutathione S-transferases and malondialdehyde in the liver of NOD mice on short-term treatment with plant mixture extract P-9801091

Changes in the concentration of glutathione S-transferases (GSTs) and malondialdehyde (MDA) were assessed in the liver of normal and diabetic NOD mice with and without treatment with the plant extract P-9801091. The plant extract P-9801091 is an antihyperglycaemic preparation containing Myrtilli folium (Vaccinium myrtillus L.), Taraxaci radix (Taraxacum of fi cinale Web.), Cichorii radix (Cichorium intybus L.), Juniperi fructus (Juniperus communis L.), Centaurii herba (Centaurium umbellatum Gilib.), Phaseoli pericarpium (Phaseolus vulgaris L.), Millefoliiherba (Achillea millefolium L.), Mori folium (Morus nigra L.), Valerianae radix (Valeriana of ficinalis L.) and Urticae herba et radix (Urtica dioica L). Hyperglycaemia in diabetes mellitus is responsible for the development of oxidative stress (via glucose auto-oxidation and protein glycation), which is characterized by increased lipid peroxide production (MDA is a lipid peroxidation end product) and/or decreased antioxidative defence (GST in the liver is predominantly an alpha enzyme, which has antioxidative activity). The catalytic concentration of GSTs in the liver was significantly reduced in diabetic NOD mice compared with normal NOD mice (p < 0.01), while the concentration of MDA showed a rising tendency (not significant). The results showed that statistically significant changes in antioxidative defence occurred in the experimental model of short-term diabetes mellitus. A 7-day treatment with P-9801091 plant extract at a dose of 20 mg/kg body mass led to a significant increase in the catalytic concentration of GSTs in the liver of diabetic NOD mice (p < 0.01) and a decrease in MDA concentration (not significant), which could be explained by its antihyperglycaemic effect.     

Aged garlic extract, garlic powder extract, S-allylcysteine, diallyl sulfide and diallyl disulfide do not interfere with the antibiotic activity of gentamicin

It was shown that aged garlic extract (AGE), garlic powder and the following garlic-derived compounds: S-allylcysteine (SAC), diallyl sulfide (DAS) and diallyl disulfide (DADS), ameliorate gentamicin (GM)-induced nephrotoxicity in rats. However, it was not established if the above mentioned extracts and compounds of garlic could interfere with the antibiotic action of GM. To address this point, AGE, garlic powder extract (GPE), SAC, DAS and DADS were assessed for their ability to interfere with the in vitro antibiotic activity of GM in Escherichia coli cultures. It was found that the above mentioned extracts and compounds of garlic were unable to decrease the antibiotic capacity of GM and even SAC, DAS and DADS alone inhibited the growth of Escherichia coli and enhanced the antibiotic effect of GM. Our data show that SAC, DAS and DADS are antibacterial compounds against E. coli and suggest that AGE, GPE, SAC, DAS and[sol ]or DADS may be administered along with GM-treatment to ameliorate GM-induced nephrotoxicity without interfering with its antibiotic activity.     

大蒜蒜酶的提取纯化及其酶学性质

大蒜为百合科多年生草本植物,可药食两用,一直受到青睐。大蒜主要具有抗感染、防治心脑血管疾病、抗肿瘤等功效。目前公认大蒜的功效成分是蒜酶(alliinase,EC4.4.1.4)催化裂解蒜氨酸(alliin)产生的大蒜辣素(al-licin)、阿藿烯(ajoene)等系列含硫有机化合物。主要从大蒜的功效成分、蒜酶的提取纯化、蒜酶的酶学性质及展望等方面综述了蒜酶的研究进展。表明大蒜制剂以其独特的显著功效,必将为人类的健康做出巨大贡献。     

清宫寿桃丸抗衰老机制分析

清宫寿桃丸由鹿肾、狗肾、人参、麦冬、枸杞子、当归、益智、酸枣仁、分心木、蚕砂等组成,原名增寿蟠桃丸,系清朝乾隆皇帝常用秘方。20世纪80年代清宫医案研究室陈可冀院士等挖掘出此方,天津达仁堂制药厂将其开发上市。该方有补肾生精、益元强壮功效,用于肾虚衰老所致头晕疲倦,