Phytochemicals of Periploca aphylla Dcne. ameliorated streptozotocin-induced diabetes in rat.

BACKGROUND: Periploca aphylla is used by local population and indigenous medicine practitioners as stomachic, tonic, antitumor, antiulcer, and for treatment of inflammatory disorders. The aim of this study was to evaluate antidiabetic effect of the extract of P. aphylla and to investigate antioxidant and hypolipidemic activity in streptozotocin (STZ)-induced diabetic rats. METHODS: The present research was conducted to evaluate the antihyperglycemic potential of methanol extract of P. aphylla (PAM) and subfractions n-hexane (PAH), chloroform (PAC), ethyl acetate (PAE), n-butanol (PAB), and aqueous (PAA) in glucose-overloaded hyperglycemic Sprague-Dawley rats. Based on the efficacy, PAB (200 mg/kg and 400 mg/kg) was tested for its antidiabetic activity in STZ-induced diabetic rats. Diabetes was induced via intraperitoneal injection of STZ (55 mg/kg) in rat. Blood glucose values were taken weekly. HPLC-DAD analysis of PAB was carried out for the presence of various polyphenols. RESULTS: HPLC-DAD analysis of PAB recorded the presence of rutin, catechin, caffeic acid, and myricetin. Oral administration of PAB at doses of 200 and 400 mg/kg for 21 days significantly restored (P < 0.01) body weight (%) and relative liver and relative kidney weight of diabetic rats. Diabetic control rats showed significant elevation (P < 0.01) of AST, ALT, ALP, LDH, total cholesterol, triglycerides, LDL, creatinine, total bilirubin, and BUN while reduced (P < 0.01) level of glucose, total protein, albumin, insulin, and HDL in serum. Count of blood cells and hematological parameters were altered in diabetic rats. Further, glutathione peroxidase, catalase, superoxide dismutase, glutathione reductase, and total soluble protein concentration decreased while concentration of thiobarbituric acid reactive substances and percent DNA damages increased (P < 0.01) in liver and renal tissues of diabetic rats. Histopathological damage scores increased in liver and kidney tissues of diabetic rats. Intake of PAB (400 mg/kg) resulted in significant improvement (P < 0.01) of above parameters, and results were comparable to that of standard drug glibenclamide. CONCLUSION: The result suggests the antihyperglycemic, antioxidant, and anti-inflammatory activities of PAB treatment in STZ-compelled diabetic rat. PAB might be used as new therapeutic agent in diabetic patients to manage diabetes and decrease the complications.

Hepatoprotective potential of Fagonia olivieri DC. against acetaminophen induced toxicity in rat.

BACKGROUND: Fagonia olivieri (DC) being used for the treatment of diabetes, cancer, fever and claimed to be effective in many other stress related disorders. In this study we have evaluated the F. olivieri whole methanol extract and its derived fractions for various in vitro and in vivo antioxidant studies. METHODS: The crude methanol extract of the whole plant of F. olivieri (FOM) and its derived fractions; n-hexane (FOH), chloroform (FOC), ethyl acetate (FOE), n-butanol (FOB) and aqueous (FOA) were evaluated for the total phenolic and flavonoid content and in vitro antioxidant abilities. The antioxidant effect of FOM was determined by acetaminophen-induced hepatotoxicity in Sprague-Dawley (Rattus novergicus) male rats. The methanol/fractions were also analysed by HPLC analysis for the presence of polyphenolics. RESULTS: The total phenolic content of the samples ranged from 19.3 ± 0.529 to 106.2 ± 0.892 mg GAE/g extract while total flavonoid content 16.2 ± 0.881 to 50.1 ± 1.764 mg RTE/g extract, respectively. FOA showed highest radical scavenging activity for DPPH (IC50 = 55.2 ± 1.212 µg/ml), ABTS (IC50 = 90.2 ± 1.232 µg/ml) superoxide (IC50 = 37.1 ± 0.643 µg/ml) and for H2O2 (IC50 = 64 ± 1.463 µg/ml). FOE exhibited the highest antioxidant activities for phosphomolybdenum (IC50 = 78.2 ± 0.883 µg/ml) and for hydroxyl radical scavenging (IC50 = 82 ± 2.603 µg/ml). HPLC analysis of FOM and its derived fractions showed the presence of rutin, catechin and gallic acid. Elevated levels of AST, ALT, ALP, LDH and lipid profile in serum and lipid peroxidation and DNA damages in liver; while decreased activity level of CAT, SOD, GSH-Px, GR and reduced glutathione (GSH) concentration induced with acetaminophen in rat were reverted towards the control group with co-administration of FOM. CONCLUSION: Our results showed that F. olivieri is a potential source of natural antioxidants, which justifies its use in folklore medicine.

Assessment of phytochemicals, antimicrobial and cytotoxic activities of extract and fractions from Fagonia olivieri (Zygophyllaceae).

BACKGROUND: In Pakistan Fagonia olivieri (Zygophyllaceae) is commonly used in the indigenous system of medicine for treatment of conditions like diabetes, cancer, fever, asthma, toothache, stomach troubles and kidney disorders. This study evaluated the crude methanol extract of F. olivieri (FOM) and its derived fractions for their antimicrobial and cytotoxic activities as well as the classes of phytochemical. METHODS: Dried powder of whole plant of F. olivieri was extracted with methanol (FOM) and the resultant was fractionated to give n-hexane fraction (FOH), chloroform fraction (FOC), ethyl acetate fraction (FOE), n-butanol fraction (FOB) and residual aqueous fraction (FOA). Methanol extract and its derived fractions were subjected to phytochemical screening using standard procedures. Also the extract and fractions were assayed for antibacterial, antifungal and cytotoxic activities using agar well diffusion technique, agar tube dilution method and brine shrimps lethality test, respectively. RESULTS: The results obtained for phytochemical analysis indicate the presence of saponins and alkaloids in all the tested extract and fractions while anthraquinones were not detected. The results showed that all the bacterial strains tested in this study were susceptible to at least one of the fractions tested. However, FOE and FOB were the best antibacterial fractions and showed antibacterial activity against maximum number of bacterial strains. The results showed that Escherichia coli was the most sensitive bacterium while Bordetella bronchiseptica and Enterobacter aerogenes were less susceptible against various fractions. Maximum percent inhibition for growth was recorded for the fungus Aspergillus flavus with FOE whereas growth of Aspergillus fumigatus and Fusarium solani was inhibited by FOM and its all derived fractions. Minimum LC50 (24.07 mg/L) for brine shrimp assay was recorded for FOE followed by LC50 of FOC (26.1 mg/L) and FOB (30.05 mg/L) whereas maximum LC50 was exhibited by FOH (1533 mg/L). CONCLUSION: These results indicated the use of F. olivieri to treat infections with emphasis to isolate and characterize the active principle responsible for antibacterial, antifungal and cytotoxic activities and its exploitation as therapeutic agent.

Investigation on flavonoid composition and anti free radical potential of Sida cordata.

BACKGROUND: Sida cordata, a member of Family Malvaceae is used in folk medicine for various ailments including liver diseases. In this study we investigated, its flavonoid constituents, in vitro antioxidant potential against different free radicals and hepatoprotection against carbon tetrachloride (CCl4)-induced liver damage in rat. METHODS: Dried powder of S. cordata whole plant was extracted with methanol and the resultant (SCME) obtained was fractionated with escalating polarity to obtain n-hexane fraction (SCHE), ethyl acetate fraction (SCEE), n-butanol fraction (SCBE) and the remaining soluble portion as aqueous fraction (SCAE). Diverse in vitro antioxidants assays such as DPPH, H2O2, •OH, ABTS, ß-carotene bleaching assay, superoxide radical, lipid peroxidation, reducing power, and total antioxidant capacity were studied to assess scavenging potential of methanol extract and its derived fractions. On account of marked scavenging activity SCEE was selected to investigate the hepatoprotective potential against CCl4 induced toxicity in Sprague-Dawley male rats by assessing the level of serum markers (alkaline phosphatase, alanine transaminase, aspartate transaminase, lactate dehydrogenase, bilirubin, and γ-glutamyltransferase) and of liver antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), glutathione-S-transfers (GST), glutathione reductase (GSR), glutathione peroxidase (GSH-Px), and reduced glutathione (GSH) and lipid peroxidation (TBARS). Histology of the liver was performed to study alteration in histoarchitecture. Existence of active flavonoids was established by thin layer chromatographic studies. RESULTS: Considerable amount of flavonoid and phenolic contents were recorded in the methanol extract and its derived fractions. Although the extract and all its derived fractions exhibited good antioxidant activities however, the most distinguished scavenging potential was observed for SCEE. Treatment of SCEE decreased the elevated level of serum marker enzymes induced with CCl4 administration whereas increased the activity of hepatic antioxidant enzymes (CAT, SOD, POD, GST, GSR and GSH-Px). Hepatic concentration of GSH was increased while lipid peroxidation was decreased with SCEE administration in CCl4 intoxicated rats. Presence of apigenin with some unknown compounds was observed in SCEE by using thin layer chromatography. CONCLUSIONS: These results revealed the presence of some bioactive compound in the ethyl acetate fraction, confirming the utility of S. cordata against liver diseases in folk medicine.

Hepatoprotection with a chloroform extract of Launaea procumbens against CCl4-induced injuries in rats.

BACKGROUND: Launaea procumbens (Asteraceae) is used as a folk medicine to treat hepatic disorders in Pakistan. The effect of a chloroform extract of Launaea procumbens (LPCE) was evaluated against carbon-tetrachloride (CCl4)-induced liver damage in rats. METHODS: To evaluate the hepatoprotective effects of LPCE, 36 male Sprague-Dawley rats were equally divided into six groups. Animals of group 1 (control) had free access to food and water. Group II received 3 ml/kg of CCl4 (30% in olive oil v/v) via the intraperitoneal route twice a week for 4 weeks. Group III received 1 ml of silymarin via gavage (100 mg/kg b.w.) after 48 h of CCl4 treatment whereas groups IV and V were given 1 ml of LPCE (100 and 200 mg/kg b.w., respectively) after 48 h of CCl4 treatment. Group VI received 1 ml of LPCE (200 mg/kg b.w.) twice a week for 4 weeks. The activities of the antioxidant enzymes catalase, peroxidase (POD), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutathione S-transferase (GST), glutathione reductase (GSR), glutathione (GSH) and lipid peroxidation (thiobarbituric acid reactive substances (TBARS)) were measured in liver homogenates. DNA damage, argyrophilic nucleolar organizer regions (AgNORs) counts and histopathology were studied in liver samples. Serum was analyzed for various biochemical parameters. Phytochemical composition in LPCE was determined through high-performance liquid chromatography (HPLC). RESULTS: LPCE inhibited lipid peroxidation, and reduced the activities of aspartate transaminase, alanine transaminase, alkaline phosphatase, and lactate dehydrogenase in serum induced by CCl4. GSH contents were increased as were the activities of antioxidant enzymes (catalase, SOD, GST, GSR, GSH-Px) when altered due to CCl4 hepatotoxicity. Similarly, absolute liver weight, relative liver weight and the number of hepatic lesions were reduced with co-administration of LPCE. Phyochemical analyses of LPCE indicated that it contained catechin, kaempferol, rutin, hyperoside and myricetin. CONCLUSION: These results indicated that Launaea procumbens efficiently protected against the hepatotoxicity induced by CCl4 in rats, possibly through the antioxidant effects of flavonoids present in LPCE.

Antioxidant, anti-inflammatory and hypoglycemic effects of Fagonia olivieri DC on STZ-nicotinamide induced diabetic rats - In vivo and in vitro study.

ETHNOPHARMACOLOGICAL RELEVANCE: Fagonia olivieri (Zygophyllaceae) is used in Pakistan for management of diabetes and for treatment of stress related disorders of liver and kidneys. The objective of present study was to evaluate antidiabetic activity of F. olivieri using streptozotocin-nicotinamide induced diabetic rat model. METHODS: Powder of the whole plant of F. olivieri was extracted with methanol (FOME) and the residue was fractionated with n-hexane (FOHE), chloroform (FOCE), ethyl acetate (FOCE), n-butanol (FOBE) and the remaining soluble aqueous fraction (FOAE). The extract/fractions were evaluated for in vitro inhibition of α-amylase and α-glucosidase and for the antidiabetic studies in Sprague-Dawley rats. FOAE was evaluated in 21 day chronic multiple dose study. Glibenclamide (10 mg/kg) and FOAE were used at a concentration of 200 mg/kg (164.8 µg of catechin) and 400 mg/kg (329.6 µg of catechin) in streptozotocin (STZ)-induced diabetic rats. Various serum parameters, blood cell parameters and oxidative stress induced parameters of the liver and kidneys were investigated. RESULTS: Results showed that aqueous fraction of F. olivieri (FOAE) potentially inhibited activity of pancreatic α-amylase, intestinal maltase and sucrase. Administration of FOAE (200 mg/kg; 400 mg/kg) to diabetic rats for 21 days increased body weight, hemoglobin and its indices, RBCs, platelets, lymphocytes and neutrophils against diabetic group. In serum level of glucose, transaminases, ALP, LDH, total bilirubin, creatinine and lipids decreased while insulin, total protein and albumin increased in contrast to diabetic group. Further, level of cytokines; TGF-ß1, TNF-α and IL-6 in serum and altered level of antioxidant enzymes, glutathione and lipid peroxides of liver and kidney tissues restored towards control animals. CONCLUSION: This study demonstrated that FOAE inhibited activity of digestive enzymes and exhibited antioxidant, anti-inflammatory and antihyperglycemic activity in STZ-nicotinamide prompted diabetic rats and suggested that FOAE might be used as a therapeutic agent for the management of type-2 diabetes.

Fagonia olivieri prevented hepatorenal injuries induced with gentamicin in rat.

Gentamicin is used clinically against Gram negative bacteria because of its efficacy. However, it causes renal injuries and failure at clinical dosages on account of induced oxidative stress. Fagonia olivieri is used in kidneys, liver, alimentary canal and cardiovascular disorders in local system of medicine in Pakistan. This study evaluates the protective abilities of methanol extract of F. olivieri (FOM) against the liver and renal injuries induced with gentamicin in rat. Sprague-Dawley male rats were treated with gentamicin (80mg/kg) alone or with silymarin (50mg/kg) as standard antioxidant drug. The other groups of rats were treated with gentamicin along with FOM (200mg/kg, 400mg/kg) or FOM (400mg/kg) alone. Effect of FOM was investigated on body weight, urine and serum biochemical markers, enzymatic antioxidants of renal and liver along with histopathological alterations. FOM was investigated by GC-MS for the presence of bioactive constituents. Treatment of FOM to rats ameliorated the gentamicin induced toxicity and increased the percent increase in body weight while decreased the absolute and relative weight of hepatic and kidneys. Gentamicin increased the level of specific gravity, count of RBCs and WBCs, and urobilinogen in urine; and AST, ALT, ALP, LDH, total bilirubin, total cholesterol, triglycerides and LDL in serum. Level of lipid peroxides in terms of thiobarbituric acid reactive substances (TBARS) and DNA damages increased while the GSH contents and activity level of CAT, SOD, GSH-Px and GR decreased with gentamicin in liver and kidney samples. Co-treatment of FOM, dose dependently, alleviated the injuries induced with gentamicin. Histopathological studies of liver and kidneys supported the protective abilities of FOM. GC-MS analysis indicated the presence of various compounds including hexadecanoic acid, 2-methoxy-4-vinylphenol, benzoic acid and thalicpureine in the FOM. Results of the present investigation suggested the protective potential of FOM against the oxidative injuries of gentamicin that could be attributed by phytochemicals having antioxidant and free radical scavenging abilities.

Assessment of antioxidant potential, total phenolics and flavonoids of different solvent fractions of monotheca buxifolia fruit.

OBJECTIVES: This study was conducted to investigate the antioxidant potential of methanol extract and its derived fractions (hexane, ethyl acetate, butanol, and aqueous) of fruits of Monotheca buxifolia (Falc.) Dc., a locally used fruit in Pakistan. METHODS: Dried powder of the fruit of M. buxifolia was extracted with methanol and the resultant was fractionated with solvents having escalating polarity; n-hexane, chloroform, ethyl acetate, n-butanol and the residual soluble aqueous fraction. Total phenolic and total flavonoid contents were estimated for the methanol and various fractions. These fractions were also subjected to various in vitro assays to estimate the scavenging activity for 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), superoxide, hydroxyl, hydrogen peroxide and reductive ability for ferric ions and phosphomolybdate assay. RESULTS: The n-butanol, aqueous and methanol fractions possessed high amount of phenolics and flavonoids compared with other fractions, and subsequently showed a pronounced scavenging activity on DPPH, ABTS, superoxide, hydroxyl and hydrogen peroxide radicals and had a potent reductive ability on ferric ion and phosphomolybdate assay. There was a found significant correlation between total phenolic and flavonoid contents and EC50 of DPPH, superoxide, hydrogen peroxide radical and phosphomolybdate assays, whereas a nonsignificant correlation was found with the hydroxyl radical and ABTS radical assay. CONCLUSION: M. buxifolia fruit can be used as natural antioxidant source to prevent damage associated with free radicals.

Evaluation of diverse antioxidant activities of Galium aparine.

Methanol extract and its n-hexane, ethyl acetate, butanol and aqueous fraction of Galium aparine L. (Rubiacea) were evaluated in vitro for their antioxidant capacity (DPPH, superoxide radical, phosphomolybdate assay); reducing power (ABTS, hydroxyl, hydrogen peroxide, to reduce Fe(3+) to Fe(2+) ions) and to estimate total flavonoid and phenolic contents. All the free radical generating assay models depicted differential positive scavenging activity but considerable magnitude for all the fractions. The results showed that aqueous fraction strongly scavenge the DPPH, ABTS, hydroxyl, hydrogen peroxide and superoxide radicals. A significantly high correlation coefficient existed between IC(50) values of DPPH and superoxide radical with total phenolic content and phosphomolybdate assay with total flavonoid contents, respectively. These results suggested that aqueous fraction can be a good source of antioxidant therapeutic in oxidative stress damages.