Effect of methanolic extract of Rumex hastatus roots (MRR) and its derived fractions, n-hexane (HRR), ethyl acetate (ERR), chloroform (CRR), butanol (BRR), and aqueous extract (ARR), was studied against carbon tetrachloride (CCl4) induced hepato and testicular toxicity in rats. Intraperitoneal dose of 20 percent CCl4 (0.5 ml/kg bw) was administered twice a week for eight weeks to a group of rats. Other groups were given CCl4 and various fractions of R. hastatus roots (200 mg/kg bw). CCl4 treatment depleted glutathione contents and activities of antioxidant enzymes while increased the concentration of lipid peroxides (TBARS) along with corresponding DNA injuries and histopathological damages. Supplementation with various fractions of R. hastatus roots (200 mg/kg body weight) attenuated the toxicity of CCl4 in liver and testis tissues through improvement in the serological, enzymatic, and histological parameters towards the normal. Posttreatment of R. hastatus roots (200 mg/kg body weight) also reversed the alteration in reproductive hormonal secretions and DNA damages in CCl4 treated rats. The results clearly demonstrated that R. hastatus treatment augments the antioxidants defense mechanism and provides the evidence that it may have a therapeutic role in free radical mediated diseases.
Mallotus philippensis (Lam.) Muell. Arg. is a well known medicinal plant of Asia and Australia. Various compounds from different aerial parts of the plant have been reported possessing potent pharmacological, antiviral, antibacterial and cytotoxic activities. We were interested to determine the effects of some root extracts from M. philippensis on human promyelocytic leukemia HL-60 cell proliferation, cell cycle regulators and apoptosis in order to investigate its anti-leukemic potential.
Root extract of M. philippensis was initially extracted in organic solvents, hexane, ethyl acetate, and n-butanol. The hexane extract showed highest toxicity against p53-deficient HL-60 cells (IC50 1.5 mg dry roots equivalent/ml medium) after 72 h and interestingly, inhibition of cell proliferation was preceded by the upregulation of the proto-oncogenes Cdc25A and cyclin D1 within 24 h. The hexane extract induced 18% apoptosis after 48 h of treatment. Chemical composition of the hexane extract was analyzed by GC-MS and the 90% fragments were matched with polyphenolic compounds.
The present study confirms that the hexane fraction of M. philippensis root extract possesses anti-leukemic activity in HL-60 cells. The polyphenols were the main compounds of the hexane extract that inhibited proliferation and induced apoptosis.
Mallotus philippensis; HL-60 cells; GC-MS; Cdc25A; Cyclin D1
Carissa opaca leaves are traditionally used in the treatment of male dysfunction and hormonal disorder as well as in oxidative stress in Pakistan and Asia. The present study was designed to assess the protective effects of methanolic extract of Carissa opaca leaves (MLC) on carbon tetrachloride (CCl4)-induced reproductive stress in male rats and bioactive constituents responsible for the activity.
CCl4 was induced in 42 male rats for eight weeks and checked the protective efficacy of methanolic extract of Carissa opaca leaves at various hormonal imbalances, alteration of antioxidant enzymes, DNA fragmentation levels and lipid peroxidation caused testicular fibrosis in testis while High performance Liquid Chromatography (HPLC) was used for detection of bioactive components.
HPLC characterization revealed the presence of isoquercitin , hyperoside , vitexin , myricetin and kaempherol. CCl4 caused significant alteration in the secretion of reproductive hormones. Activity of antioxidant enzymes viz; catalase, superoxide dimutase and phase II metabolizing enzymes including glutathione peroxidase, glutathione reductase and reduced glutathione was decreased while DNA fragmentation, hydrogen per oxide contents and thiobarbituric acid reactive substances (TBARS) were increased with CCl4 treatment. Co-administration of 100 mg/kg and 200 mg/kg b.w. MLC effectively ameliorated the alterations in the biochemical markers; hormonal and molecular levels.
Protective effects of methanolic extract of Carissa opaca against CCl4−induced antioxidant and hormonal dysfunction which might be due to bioactive compound present in extract.
CCl4; Carissa opaca; TBARS; LH; DNA fragmentation
Stress caused by free radicals accumulation result into many hazardous diseases. A number of investigations are focusing to find out the plant oriented natural antioxidant moieties. The basic aim of this research was to investigate the antioxidant potential, total Phenolic and flavonoids contents and photochemical screening of the crude methanol extract and its derived various fractions Dicliptera roxburghiana of Acanthaceae family.
Crude methanol extract of aerial parts of Dicliptera roxburghiana (DRME) was partitioned in to n-hexane (DRHF), chloroform (DRCF), ethyl acetate (DREF), n-butanol (DRBF) and the remaining soluble portion as residual aqueous fraction (DRAF). We evaluated the antioxidant activities of the extract and various fractions through different analytical methods such as DPPH, superoxide anion, ABTS, H2O2, hydroxyl radical and phosphomolybdate radical inhibition. In vitro lipid peroxidation and reducing power of the plant was also analyzed. Total flavonoid and phenolic contents of the extract and all fractions were also quantified. Plant was also subjected for preliminary phytochemical screening to confirm the presence or absence of various constituents in the plant.
Phytochemical screening confirmed the presence of flavonoids, phenolics, tannins, alkaloids, saponins, terpenoids and coumarines. Quantitative analysis revealed the maximum amount of total phenolic and flavonoid contents in DRME while lowest in DRHF. Methanol extract, DREF, DRCF and DRBF exhibited promising antioxidant potential for DPPH, ABTS, H2O2, phosphomolybdate, superoxide anion and hydroxyl radical scavenging capabilities, while these were not appreciable for DRHF and DRAF. All fractions except DRHF and DRAF possess strong reducing power ability and showed appreciable lipid peroxidation inhibition.
These research investigations revealed that Dicliptera roxburghiana is a potent source of natural antioxidants. Hence the plant can be used for management of different stress and anxiety related ailments.
Dicliptera roxburghiana; Lipid peroxidation; Total flavonoids; Antioxidants
Sonchus asper is traditionally used in Pakistan for the treatment of reproductive dysfunction and oxidative stress. The present investigation was aimed to evaluate chloroform extract of Sonchus asper (SACE) against potassium bromate-induced reproductive stress in male rats.
20 mg/kg body weight (b.w.) potassium bromate (KBrO3) was induced in 36 rats for four weeks and checked the protective efficacy of SACE at various hormonal imbalances, alteration of antioxidant enzymes, and DNA fragmentation levels. High performance chromatography (HPLC) was used for determination of bioactive constituents responsible.
The level of hormonal secretion was significantly altered by potassium bromate. DNA fragmentation%, activity of antioxidant enzymes; catalase (CAT), peroxidase (POD), superoxide dismutase (SOD) and phase II metabolizing enzymes viz; glutathione reductase (GSR), glutathione peroxidase (GSHpx), glutathione-S-tansase (GST) and reduced glutathione (GSH) was decreased while hydrogen per oxide contents and thiobarbituric acid reactive substances (TBARS) were increased with KBrO3 treatment. Treatment with SACE effectively ameliorated the alterations in the biochemical markers; hormonal and molecular levels while HPLC characterization revealed the presence of catechin, kaempferol, rutin and quercetin.
Protective effects of Sonchus asper vs. KBrO3 induced lipid peroxidation might be due to bioactive compound present in SACE.
Potassium bromate; Sonchus asper; Antioxidant; FSH; DNA fragmentation; TSH
Launaea procumbens is used in the treatment of oxidative stress and mental disorders. The effects of Launaea procumbens methanolic extracts (LPMEs), i.e., 100 and 200 LPME mg/kg body weight (b.w.), on cognitive performance as well as on the activity of acetylcholinesterase, and antioxidant enzymes in rat brain tissue homogenates were evaluated.
Thirty male Sprague–Dawley rats were divided equally into three groups. Rats in group I (control) were given saline (vehicle), group II received LPME (100 mg/kg b.w., p.o.), and group III were treated with LPME (200 mg/kg b.w., p.o.) in dimethyl sulfoxide (DMSO) for 7 days. Antioxidant potential was assessed by measuring the activity of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSHpx), glutathione reductase (GSR) and glutathione-S-transferase (GST) as well as lipid peroxidation and glutathione (GSH) contents in brain tissue homogenates. Activity of acetylcholinesterase (AChE) and cognitive performance were also assessed.
LPME administration reduced the levels of lipid peroxidation products (TBARS contents), increased GSH levels and enhanced the activities of SOD, CAT, GSHpx, GSR and GST. AChE activity was reduced by LPME treatment compared with untreated controls.
These findings suggested the significant impact of LPMEs on brain function. These effects could be through the antioxidant effects of the bioactive constituents present in LPME.
Launaea procumbens; GSH; AChE; Antioxidants enzymes
Rutin, a polyphenolic flavonoid, was investigated for its protective effects against the KBrO3 induced renal injuries in rat.
Group I was control (untreated), group II was given saline 0.5 ml/kg bw (0.9% NaCl), group III was administered KBrO3 (20 mg/kg bw) intragastric twice a week for four weeks. Rutin was administered to group VI (50 mg/kg bw) and Group V (70 mg/kg bw) along with KBrO3 (20 mg/kg bw) while group VI was given rutin (70 mg/kg bw) alone twice a week for four weeks. Protective effects of rutin on KBrO3-induced nephrotoxicity in rats were determined for biochemical parameter of urine, and serum, various antioxidant enzymes, DNA and histopathological damages in kidneys.
The level of urinary red blood cells, leucocytes count, specific gravity, urea, creatinine and urobilinogen was increased (P<0.01) whereas creatinine clearance was reduced. Serum level of protein, albumin, globulin, nitrite, creatinine and blood urea nitrogen (BUN) was significantly increased (P<0.01) by KBrO3. Marked histopathological lesions, elevated DNA fragmentation and AgNORs count in renal tissues was determined. Activity of antioxidant enzymes; catalase, superoxide dismutase, glutathione peroxidase, glutathione-S-transferase, glutathione reductase, and reduced glutathione contents were decreased (P<0.01) while thiobarbituric acid reactive substances were increased (P<0.01) with KBrO3 treatment in kidneys. DNA ladder assay was intimately related with the DNA fragmentation assay. Telomerase activity was found positive in the KBrO3 treated kidneys. Treatment with rutin effectively ameliorated the alterations in the studied parameters of rat. Rutin administration alone to rats did not exhibit any significant change in any of the parameters studied.
These results suggest that rutin works as an antioxidant in vivo by scavenging reactive oxygen species and this serves to prevent oxidative renal damage in rat treated with KBrO3.
Potassium bromate; Rutin; Histopathology; Renal function test; Antioxidant; DNA fragmentation; Telomerase
Sonchus arvensis is used in the treatment of various human aliments as a traditional medicine in Pakistan. In the study its various fractions are characterized for scavenging of diverse free radicals.
Results of the present study revealed that various fractions of Sonchus arvensis significantly scavenged the free radicals (DPPH·, ABTS·+, ·OH, superoxide), however its methanolic fraction is more potent than other fractions. Significant correlation was found between DPPH·, ABTS·+, superoxide radical and total antioxidant activity with total flavonoids and phenolics contents. Phytochmical analysis revealed the presence kaempferol, quercetin, orientin, rutin, hyperoside, catechin and myricetin.
From the present data it is concluded that various fractions of Sonchus arvensis significantly scavenged the free radical, which might be due the presence of polyphenolic constituent.
Sonchus arvensis; Correlation; DPPH; Total antioxidant activity; HPLC
Sonchus asper (L.) Hill, (Asteraceae) is used in Pakistan as a traditional (“folk”) medicine for the treatment of hormonal disorders and oxidative stress. The present study was aimed to evaluate the efficacy of Sonchus asper (L.) Hill, (Asteraceae) methanolic extract (SAME) on hormonal dysfunction in thyroid tissue after carbon tetrachloride (CCl4)-induced oxidative stress.
To examine the effects of SAME against the oxidative stress of CCl4 in thyroid tissue, 30 male albino rats were used. Protective effects of SAME were observed on thyroid hormonal levels, activities of antioxidant enzymes, lipid peroxidation (TBARS) and DNA damage.
Treatment with CCl4 significantly (P<0.01) reduced the levels of T3 and T4 and increased TSH levels. CCl4 exposure in rats reduced the activities of antioxidant enzymes but increased lipid peroxidation and DNA damage. Co-administration of SAME significantly (P<0.01) improved these alterations with respect to hormonal levels, activities of antioxidant enzymes and lipid peroxidation close to those seen in control rats.
These results suggest that SAME can protect thyroid tissue against oxidative damage, possibly through the antioxidant effects of its bioactive compounds.
Sonchus asper; Oxidative stress; Antioxidant enzymes; Carbon tetrachloride; T3; T4
Traditionally various human diseases of kidneys, hormonal imbalance and sexual diseases are treated with Launaea procumbens (L). In the present study protective effects of methanolic extract of Launaea procumbens (LPME) was evaluated against CCl4-induced oxidative damages in rat testis.
To examine the protective effects of Launaea procumbens on testis against oxidative stress of carbon tetrachloride in male rat, 30 male albino rats were equally divided into 5 groups (6 rats). First group was given standard diet and drinking water. Second group received CCl4 3 ml/kg intraperitoneally (30% in olive oil). Third and forth were given orally 100; 200 mg/kg b.w., in 99.8% dimethyl sulphooxide (DMSO), Launaea procumbens methanolic extracts (LPME) after 48 h of CCl4 treatment twice a week and sixth group received only LPME in DMSO at a dose of 200 mg/kg b.w., for four weeks. Protective effects of Launaea procumbens were observed on sperm concentration, motility and morphology, serum reproductive hormonal level, activity of antioxidant enzymes, lipid peroxidation (TBARS) and DNA damages.
Results of the present study revealed that treatment of CCl4 significantly (p < 0.01) reduced sperm concentration and motility comparatively to controls. Level of testosterone, luteinizing hormone and follicle stimulating hormone, were depleted markedly (p <0.01) with treatment of CCl4. In addition, CCl4 induction in rats reduced activities of antioxidant enzymes while increased lipid peroxidation and DNA damages. Co-administration of LPME significantly (p <0.01) improved these alterations in improving of hormonal level, activities of antioxidant enzymes and lipid peroxidation near to control rats.
From the results it is suggested that Launaea procumbens methanolic extract has the ability to protect testis against oxidative damages, possibly through antioxidant effects of its bioactive compounds.
Launaea procumbens; GSH; CCl4; Lipidperoxidation; DNA damages
Launaea procumbens (L.) is traditionally used in the treatment of various human ailments including pulmonary damages. The present study was arranged to evaluate the role of Launaea procumbens methanol extract (LME) against carbon tetrachloride (CCl4) induced oxidative pulmonary damages in rat.
36 Sprague–Dawley male rats (170-180 g) were randomly divided into 06 groups. After a week of acclamization, group I was remained untreated while group II was given olive oil intraperitoneally (i.p.) and dimethyl sulfoxide (DMSO) orally, groups III, IV, V and VI were administered CCl4, 3 ml/kg body weight (30% in olive oil i.p.). Groups IV, V were treated with 100 mg/kg, 200 mg/kg of LME whereas group VI was administered with 50 mg/kg body weight of rutin (RT) after 48 h of CCl4 treatment for four weeks. Antioxidant profile in lungs were evaluated by estimating the activities of antioxidant enzymes; catalase (CAT), peroxidase (POD), superoxide dismutase (SOD), glutathione-S-transferase (GST), glutathione reductase (GSR), glutathione peroxidase (GSH-Px), quinone reductase (QR) and reduced glutathione (GSH). CCl4-induced lipid peroxidation was determined by measuring the level of thiobarbituric acid reactive substances (TBARS) with conjugation of deoxyribonucleic acid (DNA) damages, argyrophilic nucleolar organizer regions (AgNORs) counts and histopathology.
Administration of CCl4 for 6 weeks significantly (p < 0.01) reduced the activities of antioxidant enzymes and GSH concentration while increased TBARS contents and DNA damages in lung samples. Co-treatment of LME and rutin restored the activities of antioxidant enzymes and GSH contents. Changes in TBARS concentration and DNA fragmentation were significantly (p < 0.01) decreased with the treatment of LME and rutin in lung. Changes induced with CCl4 in histopathology of lungs were significantly reduced with co-treatment of LME and rutin.
Results of present study revealed that LME could protect the lung tissues against CCl4-induced oxidative stress possibly by improving the antioxidant defence system.
Launaea procumbens; Lungs; CCl4; Antioxidant enzymes; TBARS; GSH
Aerva javanica (Burm. f.) Juss. ex Schult. (Amaranthacea) is traditionally used for the treatment of wound healings, cough, diarrhoea, ulcer and hyperglycaemia. The current study was aimed to fractionate and isolate bioactive compounds and ultimately to evaluate their anti-ulcereogenic potential.
In order to achieve these aims, the fractionation, purifications and then biological potential determination of the isolated compounds was carried out. For purification purpose, initially extraction of the plant material was done with aqueous MeOH in the order of increasing polarity by using solvent-solvent extraction method. Phytochemical analysis revealed the presence of three compounds, 3-hydroxy-4 methoxybenzaldehyde (1), ursolic acid (2) and (E)-N-(4-hydroxy-3-methoxyphenethyl)-3-(4-hydroxy-3-ethoxyphenyl) acryl amide (3). Inhibition of urease activity of various fractions revealed that ethyl acetate fraction showed significant activity (P <0.05) as compared to other fractions. (E)-N-(4-hydroxy-3-methoxyphenethyl)-3-(4-hydroxy-3-ethoxyphenyl) acryl amide (3) showed marked anti ulcer activity (P <0.05).
These results suggested the mild potential of A. javanica against ulcer.
Aerva javanica; Urease activity; Ethyl acetate fraction; Solvent-solvent extraction
Launaea procumbens (LP) has been used as a food supplement in Pakistan. In this study methanolic crude extract (LPME) of the whole plant and its different fractions; n-hexane (LPHE); ethyl acetate (LPEE) and chloroform (LPCE) were studied for the determination of total flavonoid and phenolics contents along with multifaceted in vitro scavenging assays.
Considerable amount of flavonoid and phenolics contents were found in all the fractions. Methanol and chloroform fraction exhibited efficient scavenging of DPPH·, ABTS·+, ·OH, superoxide, lipid peroxide and nitric oxide free radicals. Significant correlation was found between DPPH·, ABTS·+, superoxide radical, β-carotene bleaching restraint and phosphomolybdenum assay with total flavonoids and phenolics contents. High performance chromatography (HPLC) of LPME revealed the presence of vitexin, orientin, rutin, hyperoside, catechin and myricetin.
These results reveal the presence of bioactive compounds in LPME, which might be contributed towards the various in vitro scavenging.
Launaea procumbens; Scavenging of DPPH-free radicals; Superoxide radicals; HPLC; Flavonoids
Sonchus asper (SA) is traditionally used as a folk medicine to treat mental disorders in Pakistan. The aim of this study was to investigate the effect of polyphenolic rich methanolic fraction of SA on cognitive performance, brain antioxidant activities and acetylcholinesterase activity in male rats.
30 male Sprague–Dawley rats were equally divided into three groups in this study. Animals of group I (control) received saline (vehicle), group II received SA (50 mg/kg) body weight (b.w.), and group III treated with SA (100 mg/kg b.w.,) orally in dimethyl sulphoxide (DMSO) for 7 days. The effect of SA was checked on rat cognitive performance, brain antioxidatant and acetylcholinesterase activities. Evaluation of learning and memory was assessed by a step-through a passive avoidance test on day 6 after two habituation trials and an initial acquisition trial on day 5. Antioxidant potential was determined by measuring activities of superoxide dismutase (SOD), catalase (CAT), contents of thiobarbituric acid reactive substances (TBARS) and reduced glutathione (GSH) in whole-brain homogenates. Acetylcholinesterase (AChE) activity was determined by the colorimetric method.
Results showed that 100 mg/kg b.w., SA treated rats exhibited a significant improvement in learning and memory (step-through latency time). SA administration reduced lipid peroxidation products and elevated glutathione levels in the SA100-treated group. Furthermore, salt and detergent soluble AChE activity was significantly decreased in both SA-treated groups. Short-term orally supplementation of SA showed significant cognitive enhancement as well as elevated brain antioxidant enzymes and inhibited AChE activity.
These findings stress the critical impact of Sonchus asper bioactive components on brain function.
Sonchus asper; Cognitive performance; Acetylcholinesterase activity; Antioxidant enzymes
Sonchus asper (SA) is traditionally used for the treatment of various ailments associated with liver, lungs and kidneys. This study was aimed to investigate the therapeutic potential of nonpolar (hexane, SAHE; ethyl acetate, SAEE and chloroform, SACE) and polar (methanol, SAME) crude extracts of the whole plant.
To achieve these goals, several parameters including free-radical (DPPH•, ABTS•+, H2O2 and •OH) scavenging, iron chelating activity, scavenging of superoxide radicals, total flavonoids and total phenolic content (TPC) were examined.
The SA extracts presented a remarkable capacity to scavenge all the tested reactive species with IC50 values being found at the μg ⁄ ml level. The SAME was shown to have the highest TPCs while lowest IC50 values for the DPPH•, ABTS•+ radical scavenging capacities and iron chelating scavenging efficiency, moreover, SAME had best activities in scavenging of superoxide radicals and hydrogen peroxide as well as potently scavenged the hydroxyl radicals.
These results suggest the potential of S. asper as a medicine against free-radical-associated oxidative damage.
Sonchus asper; Antioxidant activities; Solvent extraction; Phenolics