Isolation and chromatographic analysis of bioactive triterpenoids from the bark extract of Cariniana domestica (Mart) Miers

The Lecythidaceae family is composed of 25 genera and 400 species that occur in the form of trees with a pantropical distribution. The genera Cariniana belongs to the family Lecythidaceae, and there are few reports considering these species. In this work, fractionation of the dichloromethane fraction obtained from the aqueous extract of the stem bark of Cariniana domestica (Mart) Miers, popularly known as Jequitibá-roxo, led to the isolation of two mixtures of triterpenoids: lupeol and β-amyrin and β-sitosterol and stigmasterol. The structures of the isolated compounds were elucidated by spectroscopic (NMR) and chromatographic (GC-MS) techniques as well as literature data comparisons. β-Sitosterol, stigmasterol, lupeol and β-amyrin were quantified in dichloromethane fraction by high-performance liquid chromatography (HPLC/DAD). The dichloromethane fraction was also investigated for antioxidant and antifungal activities. The isolated compounds and their biological activities are reported for the first time for the species C. domestica.     

Hepatoprotective constituents of Torilis radiata Moench (Apiaceae)

An investigation of the aqueous ethanolic extract (AE) of the aerial parts of Torilis radiata Moench yielded two triterpenes (lupeol acetate (1) and α-amyrin (2)), a sterol (spinasterol (3)) from its n-hexane fraction (HF), a flavone (acacetin (4)), a coumarin (scopoletin (5)), a phenolic acid (ferulic acid (6)) from the chloroform fraction (CF) and a flavone glycoside (luteolin-7-O-glucoside (7)) from the n-butanol fraction (BF). The hepatoprotection of the AE and its fractions was assessed in terms of the reduction in histological damage, accompanied by restoration of the liver enzymes (alanine amino transferase (ALT), aspartate amino transferase (AST), lactate dehydrogenase (LDH)), a reduction in the inflammatory markers (tumour necrosis-α (TNF-α), nitric oxide (NO), N-acetyl-β-D-glucosaminidase (NAG) and myloperoxidase (MPO) in serum) and restoration of the oxidant balance by decreasing the serum and hepatic malondialdehyde (MDA) levels, along with increasing the activity of hepatic catalase (CAT), glutathione peroxidase (GSHPx) and the non-enzymatic antioxidant glutathione (GSH).     

Inhibitory Effect of Lupeol on MMPs Expression using Aged Fibroblast through Repeated UVA Irradiation

In this study, the aged dermal fibroblast model was constructed by repeated irradiation with UV light and the effect of lupeol, a triterpenoid, on anti‐aging was confirmed. SA‐β‐galactosidase (SA‐β‐gal) stained aged cells increased by about 40% and expression of p‐p53, p21, p16 and MMPs (MMP‐1, ‐2, ‐3) increased in aged fibroblast. As an efficacy result, the treatment of lupeol on aged fibroblast induced by UVA repeated irradiation showed a dose‐dependent reduction of SA‐β‐gal stained aged cells, the expression of p‐p53, p21, p16 and inhibition of MMPs. Interestingly, lupeol increased dephosphorylation of p‐ERK in repeated UV irradiated conditions. Additionally, lupeol compensated MMPs expression when p‐ERK phosphorylation was inhibited by p‐ERK inhibitor PD98059. Thus, these results showed that lupeol has a possible effect on MMPs expression using inhibition of the p‐ERK pathway. Taken together, we confirmed that lupeol inhibits senescence through inhibiting MMP‐1, ‐2, ‐3 as well as p‐p53, p21 and p16 expression and SA‐β‐gal activity in repeated UVA‐irradiated senescent FB models, therefore suggesting that lupeol may be useful as an anti‐aging agent.     

Green synthesis of gold nanoparticles from Fritillaria cirrhosa and its anti-diabetic activity on Streptozotocin induced rats

To evaluate the anti-diabetic effect of Fritillaria cirrhosa gold nanoparticles on Streptozotocin (STZ) stimulated diabetic preclinical models. The albino rats of either sex were equally distributed to five different groups. Group-I represented as Control; Group-II represented as diabetic control (STZ alone); Group-III represented as 10 mg/kg body weight of Fritillaria cirrhosa gold nanoparticles + diabetes; Group-IV represented as 20 mg/kg body weight of Fritillaria cirrhosa gold nanoparticles + diabetes; Group-V represented as 0.1 mg/kg body weight of glibenclamide + diabetes. The animals were killed after the experimental period. The blood and organs samples were gathered and stored for the additional investigations. The gold nanoparticles were inspected via the UV spectrophotometer, HR-TEM, XRD and FT-IR techniques. The bodyweight, kidney and liver weight were tabulated. The food and water consumption were monitored in all the experimental rats. The serum markers, hepatic markers and renal markers were quantified in both normal and investigational rats. The lipid peroxidation and antioxidant status were quantified in the control and experimental rats. The histopathological alterations were also studied in all the experimental animals. The standard drug glibenclamide was used to compare the synthesized AuNPs. The study revealed that the AuNPs treatment restores the serum, hepatic and renal marker in the STZ-challenged diabetic rats. The AuNPs treatment modulates the antioxidants level and decreased the lipid peroxidation by its antioxidant properties. The pathology results revealed that the AuNPs treatment induces the regeneration of islets cells of pancreas in the experimental rats. The research study proved that the Fritillaria cirrhosa AuNPs exerts anti-diabetic properties.     

Effect of ethanol extract of Sphaeranthus indicus on cisplatin-induced nephrotoxicity in rats

Cisplatin is an anticancer drug extensively used against a variety of cancers. Cisplatin chemotherapy is found to manifest dose-dependent nephrotoxicity. Depletion of the renal antioxidant defence system has been suggested to be the main cause of cisplatin-induced nephrotoxicity. The purpose of this study is to investigate whether the ethanol extract of entire plant of Sphaeranthus indicus could reduce the intensity of toxicity in albino rats. Nephrotoxicity was assessed by determining the serum creatinine and urea levels and renal antioxidant status in rats after cisplatin administration (12?mg?kg(-1)?body weight, i.p.). The ethanol extract of S. indicus (150 and 300?mg?kg(-1)?body weight) was administered orally from the sixth day onwards for 10 days after cisplatin administration. The extract significantly reduced the elevated serum creatinine and urea levels. Renal antioxidant defence systems, such as superoxide dismutase, catalase, glutathione peroxidase activities and reduced glutathione level that are depleted by cisplatin therapy were restored to normal by treatment with the extract. Cisplatin-induced lipid peroxidation was also found to be markedly reduced by treatment with the extract. These results suggest that S. indicus has protective effect against cisplatin-induced nephrotoxicity, which may be attributed to its antioxidant potential.     

Antioxidants Isolated from Elaeagnus umbellata (Thunb.) Protect against Bacterial Infections and Diabetes in Streptozotocin-Induced Diabetic Rat Model

The increase in resistance of microbes against conventional drugs is currently a hot issue, whereas diabetes is another main cause of mortalities encountered throughout the world after cancer and heart attacks. New drug sources in the form of plants are investigated to get effective drugs for the mentioned diseases with minimum side effects. Elaeagnus umbellata Thunb. is a medicinal plant used for the management of stress related disorders like diabetes and other health complications. The active constituents of the chloroform extract derived from E. umbellata berries was isolated by silica gel column chromatography which were identified as morin, phloroglucinol, and 1-hexyl benzene through various spectroscopic techniques (electron ionization mass spectrometry, 1H-NMR, and 13C-NMR spectroscopy). The possible protective effects (antioxidant, antibacterial, and antidiabetic activity) of isolated compounds were evaluated using reported methods. Morin exhibited strong in vitro antiradical potential against DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2′-azinobis-3-ethylbenzothiazoline-6-sulfonic acid) radicals along with prominent antibacterial activities against selected bacterial strains (Escherichia coli, Bacillus cereus, Salmonella typhi, Klebsiella pneumonia, Pseudomonas aeruginosa and Proteus mirabilis). Among the isolated compounds the more potent one (morin) was assessed for its in vivo antidiabetic potential in streptozotocin-induced diabetic rat model. The in vivo effects observed were further confirmed in ex vivo experiments where the effect of isolated compound on antioxidant enzyme like glutathione peroxidase (GPx), total content of reduced glutathione (GSH), % DPPH inhibition, and the lipid peroxidation MDA (Malondialdehyde) level in pancreatic tissues homogenates were evaluated. In vivo morin at tested doses (2, 10, 15, 30 and 50 mg/kg body weight) significantly restored the alterations in the levels of fasting blood glucose level and body weight loss along with significant decrease in levels of cholesterol, triglycerides, low density lipoprotein, HbA1c level, and significantly increased the high-density lipoprotein in diabetic rats. Morin also effectively ameliorated the hepatic enzymes, and renal functions like serum creatinine. Morin significantly increased the antioxidant enzyme like GPx activity, GSH content, and % DPPH inhibition activity, while reduced the lipid peroxidation MDA (malondialdehyde) level in pancreatic tissues homogenates, and modification of histopathological changes in diabetic rats. Morin exhibited high antioxidant, antibacterial, and antidiabetic potentials as compared to phloroglucinol and 1-hexyl benzene, that could, therefore, be considered as a promising therapeutic agent to treat diabetes mellitus and bacterial infections.     

细胞代谢组学用于羽扇豆醇干预人乳腺癌细胞MCF-7的机理探究

应用基于气相色谱-质谱联用（GC-MS）的代谢组学方法结合细胞周期实验,研究羽扇豆醇体外抑制人乳腺癌细胞MCF-7增殖的作用机理。代谢组学的研究结果表明：通过正交偏最小方差判别分析（OPLS-DA）可以很好地区分羽扇豆醇作用的MCF-7细胞代谢谱与对照组细胞代谢谱,模型参数为：R²Ycum=0.988,Q²Ycum=0.964。VIP（variable importance in the projection）值大于1的差异代谢物进一步用t检验进行单位分析,选择t<0.05（VIP>1）的代谢物作为羽扇豆醇作用组的生物标志物,得到琥珀酸、磷酸、亮氨酸、异亮氨酸等11种代谢差异物。结合羽扇豆醇将细胞周期抑制在G1期这一现象,推测羽扇豆醇可能是主要抑制了三羧酸循环中的琥珀酰辅酶A的生成和底物磷酸化生成ATP的反应来抑制MCF-7细胞的增殖。本实验从代谢组学角度为乳腺癌抗肿瘤机制提供新的线索。     

Bioactive phenolic amides from Celtis africana

Nine compounds have been isolated for the first time from Celtis africana, namely trans-N-coumaroyltyramine (1), trans-N-feruloyltyramine (2), trans-N-caffeoyltyramine (3), lauric acid (4), oleic acid (5), palmitic acid (6), lupeol (7), β-sitosterol (8) and oleanolic acid (9), respectively. Their structures have been elucidated by different spectroscopic techniques. The isolated compounds were screened for their antioxidant, anti-inflammatory and acetylcholinestrease enzyme inhibitory activities. Compounds 1-3 showed significant antioxidant and anti-inflammatory activities and weak to moderate acetylcholinestrease enzyme inhibition activity.     

Phytochemical constituents and antioxidant activity of Echinops albicaulis

Two thiophenes; 5-(3-buten-1-ynyl)-2,2′-bithiophene (2) and α-tertthienyl (9), two alkaloids; echinopsine (10) and echinorine (11), three flavonoids; genkwanin (5), apigenin (6), and rutin (7), two triterpenoids; lupeol acetate (1) and lupeol linoleate (4), together with 2,6,10-trimethyldodeca-2,6,10-triene (4) and β-sitosterol glucoside (8) were isolated from the aerial parts of Echinops albicaulis. Antioxidant, antimicrobial and antiprotozoal activities were evaluated. E. albicaulis aqueous methanolic extract (50, 10, and 1 mg/mL) showed significant antioxidant activity comparable to the potent antioxidant, N-acetyl cysteine, moreover, the aqueous methanolic extract (1 mg/mL) significantly reduced intracellular reactive oxygen species in active cell cultures of human peripheral blood mononuclear cells under oxidative stress more than the reference antioxidant N-acetyl cysteine. None of the isolated compounds showed antimicrobial or antiprotozoal activities at concentration up to 20 μg/mL.     

A bioactive spirolactone iridoid and triterpenoids from Himatanthus sucuuba.

Himatanthus sucuuba is an Amazonian tree with abundant, yet conflicting ethnobotanical information. Investigation of the polar and non-polar constituents led to the isolation of plumericin, a bioactive spirolactone iridoid, and four known pentacylic triterpenes: lupeol acetate, lupeol cinnamate, lupeol beta-phenyl propionate, and alpha-amyrin cinnamate.     

Correlation between glycated lipoproteins and fructosamine level in serum.

The correlation between the level of fructosamine and glycated proteins, including glycated lipoproteins, in serum from diabetic and nondiabetic subjects was studied. Assay of glycated proteins in serum was performed using an agarose gel film electrophoresis with nitroblue tetrazolium coloration. Glycated albumin correlated well with the fructosamine level in the diabetics (r = 0.83-0.92, p less than 0.01) but showed no correlation with the nondiabetics (r = 0.25-0.26). Also, a high correlation between the glycated beta-lipoprotein and fructosamine levels was observed in diabetic patients with hyperglycemia and in nondiabetic subjects with a high risk of atherogenesis (atherogenic index, low-density lipoprotein-cholesterol/high-density lipoprotein-cholesterol greater than 2.8) (r = 0.51-0.66, p less than 0.01). Nondiabetics with a high level of beta-lipoprotein, which is well known to cause high atherogenesity, showed a high level of glycated beta-lipoprotein similar to that in the diabetic groups with hyperglycemia; therefore, the high level of glycated beta-lipoprotein seems to be attributable not only to the hyperglycemia-accelerated glycation of beta-lipoprotein but also to an increase in the level of beta-lipoprotein in serum. Consequently, the present results show that the fructosamine level in serum reflects not only the glycation of albumin but also that of lipoproteins which are known to increase in diabetes mellitus.     

Investigation of antiplasmodial efficacy of lupeol and ursolic acid isolated from Ficus benjamina leaves extract

Abstract

This study emphasizes on the investigation of antiplasmodial activity of triterpenoids isolated from Ficus benjamina leaves. An unsaponified fraction of petroleum ether extract of plant leaves was subjected to silica gel column chromatography which led to the isolation of two known triterpenoids; namely ursolic acid and lupeol. These compounds were evaluated for antiplasmodial activity by schizont maturation inhibition assay using 3D7 Plasmodium strains. Both, ursolic acid and lupeol were found to exhibit significant antiplasmodial effect with an IC₅₀ value of 18?µg/ml and 3.8?µg/ml, respectively. This study further confirms the traditional role of Ficus benjamina plant in the treatment of malaria which may be attributed to ursolic acid and lupeol.     

Nephroprotective and antioxidant activities of Salacia oblonga on acetaminophen-induced toxicity in rats

Salacia oblonga, a woody climbing plant belonging to the family Celastaceae, is widely distributed in India and other southeast Asian countries. The genus Salacia have been used particularly for the treatment of diabetes, obesity, gonorrhoea, rheumatism, pruritus and asthma. Acetaminophen (APAP), used as an analgesic drug, produces liver and kidney necrosis in mammals at high doses. The aim of this study was to investigate the nephroprotective and antioxidant activities of the ethanol extract of Salacia oblonga (EESO) at the two dose levels of 250 and 500 mg/kg?bw on APAP-induced toxicity in rats. The results showed that APAP significantly increases the levels of serum urea, creatinine, and reduces levels of uric acid concentration. The EESO reduces these by increasing anti-oxidative responses as assessed by biochemical and histopathological parameters. In conclusion, our results suggest that the EESO possesses nephroprotective and antioxidant effects against APAP-induced nephrotoxicity in rats.     

糖尿病患者血清中微量元素分析

对２０例糖尿病患者血清锌、铁、锰、钙、铜含量检测发现，糖尿病组较对照组血清铁、铜高，差异显著（Ｐ〈０．０５），血清锌、锰略低（Ｐ〉０．０５），血清钙略高（Ｐ〉０．０５）。表明糖尿病可导致微量元素代谢紊乱，在治疗糖尿病时要适当补充微量元素；同时微量元素含量可作为糖尿病的一个控制指标。     

Protective Effect of Polyphenols Extract of Adlay (<em>Coix lachryma-jobi </em>L. var<em>. ma-yuen Stapf</em>) on Hypercholesterolemia-Induced Oxidative Stress in Rats

The present study examines the effect of polyphenols extract of adlay (Coix lachryma-jobi L. var. ma-yuen Stapf) (APE) on high cholesterol diet fed rats (HCD). APE was orally administrated by gavage at doses of 10, 40 and 200 mg total phenolics/kg body weight of rats once a day for 28 days. At the end of four weeks, serum triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C), and markers of oxidative stress viz., malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px) in the serum and liver of HCD and normal rats were assessed and compared. The results showed that administration of APE was significantly effective in decreasing the serum levels of TC, LDL-C and MDA, increasing the serum level of HDL-C and antioxidant capacity. In addition, oral gavage of APE could also increase the antioxidant capacity, CAT and GSH-Px activities in liver. These results suggested that APE exerted a high hypocholesterolemic and antioxidant activities, which might be characterized by a protective effect on cardiovascular health in vivo.     

Protective effects of Ocimum sanctum on lipid peroxidation and antioxidant status in streptozocin-induced diabetic rats

The antioxidant effects of Ocimum sanctum in experimental streptozocin-induced diabetic rats was evaluated in this study. Streptozocin, 55?mg?kg(-1) body weight, was injected intraperitoneally once daily for 30 days to induce diabetes mellitus in rats. Streptozocin-induced diabetic rats were orally treated with an aqueous extract of O. sanctum once daily for 30 days. After the experimental period, thiobarbituric acid reacting substances (TBARS) and antioxidant enzymes such as catalase, superoxide dismutase (SOD) and glutathione peroxidase were measured. Administration of O. sanctum to streptozocin-induced diabetic rats for 30 days significantly reduced the plasma level of TBARS and improved the status of the antioxidant enzymes catalase, SOD and glutathione peroxidase in vital organs such as the liver and kidney. These results confirmed that the Indian medicinal plant O. sanctum has a protective effect and it may be useful in controlling complications resulting from diabetes.     

Determination of chemopreventive role of Foeniculum vulgare and Salvia officinalis infusion on trichloroacetic acid-induced increased serum marker enzymes lipid peroxidation and antioxidative defense systems in rats

Today's world is increasingly seeking ways to replace the synthetic drugs with the therapeutic power of natural products. This study was designed to investigate the protective effects of Foeniculum vulgare (FV) and Salvia officinalis (SO) waters infusions against carcinogen chemical trichloroacetic acid (TCA)-exposure in rats. The chemopreventive potential of the plant infusions were evaluated by measuring levels of serum marker enzymes [aspartate aminotransferase (AST), alanin aminotransferase (ALT), creatine phosphokinase (CPK), acid phosphatase (ACP), alkaline phosphatase (ALP) and lactate dehydrogenase (LDH)], antioxidant defense systems [Reduced glutathione (GSH), glutathione reductase (GR), superoxide dismutase (SOD), glutathione-S-transferase (GST) and catalase (CAT)] and lipid peroxidation level (Malondialdehyde = MDA) in various tissues of rats. Female Sprague-Dawley rats, weighing 150-200 g, were randomly allotted into four experimental groups. While the control group (A) received only natural spring water, the treatment B group (0.2% TCA) supplied with the drinking water containing 0.2% TCA, the treatment C (TCA + FV infusion) and D (TCA + SO infusion) groups drank the drinking water containing 0.2% TCA and 2.5% the plant grains and leaves ad libitum for 50 days during experiment. At the end of the 50 days experiment, TCA and the plant's infusions caused different affect on the serum marker enzymes, tissues antioxidant defense systems and lipid peroxidation against TCA-exposed in rats with comparison to those of TCA exposed and control rats. According to the results, both TCA and TCA + plants infusions caused a significant increase in serum AST, ALT and CPK activity. Non-enzymic antioxidant GSH level significantly increased in the brain whereas reduced in the erythrocytes and kidney of TCA + FV and TCA + SO as compared to TCA group and control. While MDA content slightly increased in tissues of TCA group in comparison to those of control, significantly decreased in the brain, liver and kidney of rats of TCA + FV and TCA + SO groups as compared to TCA group and control. Antioxidative enzyme activity such as CAT and SOD significantly increased in the brain, liver and kidney tissues of TCA induced group whereas reduced the same enzymes activities as compared to TCA group. The ancillary enzyme GR activity significantly depleted in the brain and kidney of TCA + FV and TCA + SO groups in comparison to those of TCA exposed and control rats. In addition, the drug metabolizing enzyme GST activity significantly declined in the brain and kidney of TCA + FV and TCA + SO groups in comparison to those of TCA exposed and control rats, whereas, also reduced in the liver of TCA + FV and TCA + SO groups in comparison to those of TCA exposed rats. It was concluded that the levels of serum marker enzymes were found not to be decreased in plants treated groups due to hepatic damage induced by TCA. Also the four antioxidant enzymes were found to be activated in different degrees following TCA treatment and declined the activation of the enzymes the plant infusions accompanied by significant reduction in MDA concentration in the tissues. The observations, along with changes, might suggest that the both FV and SO may possess antioxidant properties during the period of a 50-day protective exposure.     

Antioxidant activity of whey protein fractions isolated by gel exclusion chromatography and protease treatment

Whey proteins were isolated from whey powder by a combination of gel exclusion chromatography and protease (pepsin or trypsin) treatment. Whey solution (6g/dl) was applied to Sephadex G-200 column chromatography and three fractions were obtained. Gel electrophoresis (SDS-PAGE) was used to identify the fractions; the first one contained immunoglobulins and bovine serum albumin, the second contained beta-lactoglobulin and alpha-lactalbumin whereas the third fraction contained small peptides. We have also subjected the whey filtrate to proteases (pepsin and trypsin). Treatment with proteases showed that beta-lactoglobulin can be obtained after hydrolysis of the second fraction with pepsin. When the whey filtrate was treated with pepsin and then applied to Sephadex G-200 column chromatography three fractions were obtained; the first one was bovine serum albumin, the second was beta-lactoglobulin and the third fraction contained small peptides. After trypsin treatment only two fractions were obtained; the first one was serum albumin and the second fraction was an alpha-lactalbumin rich fraction. We have determined the antioxidant activity of the fractions using an assay based on the measurement of superoxide radical scavenging activity. Our results showed that among the three fractions, the first fraction had the highest superoxide radical scavenging activity. Also, protease treatment of the second fraction resulted in an increase in the antioxidant activity.     

Antidiabetic activity of Terminalia sericea constituents

Diabetes mellitus is an endocrine disorder that affects more than 100 million people worldwide. South African plants namely Terminalia sericea, Euclea natalensis, Warbugia salutaris, Aloe ferox, Artemisia afra, Sclerocarya birrea, Spirostachys africana and Psidium guajava were investigated for their in vitro alpha-glucosidase and alpha-amylase properties, and antioxidant activities. Terminalia sericea stem bark extract showed the best results against alpha-glucosidase and alpha-amylase enzymes. Bioassay-guided fractionation of an acetone extract of T. sericea stem bark led to the isolation of four known compounds namely beta-sitosterol (1), beta-sitosterol-3-acetate (2), lupeol (3), and stigma-4-ene-3-one (4), in addition to two inseparable sets of mixtures of isomers [epicatechin-catechin (M1), and gallocatechin-epigallocatechin (M2). 1 and 3 showed the best inhibitory activity on alpha-glucosidase (IC50:54.5 and 66.5 microM). Bio-evaluation of the inhibitory activity of the purified compounds on alpha-amylase showed that 3 and 1 exhibited IC50 values of 140.7 and 216.02 microM, respectively against alpha-amylase. Compounds 2, M1, 3 and M2 were found to be non-toxic to Vero cells. This study is the first to report alpha-glucosidase and alpha-amylase activity of M1, M2, 2 and 4 isolated from T. sericea, which validated the traditional use of the bark of T. sericea for diabetes in South Africa.