Phytochemical screening and free radical scavenging activity of Citrullus colocynthis seeds extracts

Objective:To study the phytochemical screening of different extracts from Citrullus colocynthis(C.colocynthis)seeds extracts and to assess their antioxidant activity on the DPPH free radical scavenging.Methods:Phytochemical screening,total content of polyphenols and flavonoids of C.colocynthis seeds extracts,including a crude aqueous extract(E1),a defatted aqueous extract(E2),a hydromethanolic extract(HM),an ethyl acetate extract(EA)and a n-butanol extract(n-B)was carried out according to the standard methods and to assess their corresponding effect on the antioxidant activity of this plant.Results:None of these extracts contained detectable amount of alkaloid,quinone,antraquinone,or reducing sugar.Catechic tannins and flavonoids were abundant in E1,MM and EA,whilst terpenoids were abundantly present in E1 and n-B but only weekly in HM.Coumarins were found in E2,EA and n-B.Polyphenols,expressed as gallic acid equivalent,amounted,per 100 g plant matter,to 329,1002 and 150 mg in EA,HM an E1 respectively.Flavonoids,expressed as catechin equivalent,amounted.per 100 g plant matter to 620,241 and 94 mg in EA,HM and E1 respectively.Comparable values were found in n-B and E1,with lower values in E2.Quercetin,myricetin and gallic acid were found in the EA and HM extracts by thin layer chromatography,The antioxidative effect of these extracts yielded,when tested at a concentration of 2000μg/mL in a 1,1-diphenyl-2-picrylhydrazyl assay,a reducing percentage of 88.8%with EA,74.5%with HM and 66.2%with E1,and corresponding IC_(50)of 350,580and 500μg/mL as compared to 1.1μg/mL for ascorbic acid.Conclusions:These qualitative and quantitative analytical data document the presence in C.colocynthis extracts of such chemical compounds as flavonoids responsible for the antioxidant activity,as well as other biological activities of this plant.

Hepatoprotection: A Hallmark of <i>Citrullus colocynthis</i>L. against Paracetamol Induced Hepatotoxicity in Swiss Albino Rats

Objective: To demonstrate the in-vivo hepatoprotective effect of the ethanolic extracts of Citrullus colocynthis (Linn.) against paracetamol induced hepatotoxicity in albino rats. Animal Model: Swiss Albino rats of either sex were used, divided into six groups with six in each group. Group 1-Normal control: The animals were maintained under normal control, which were given distilled water only. Group 2-Induction of hepatotoxicity: The animals received paracetamol 500 mg/kg b.w. (p.o) every 72 h for 10 Days. Groups 3 to 5: Animals received ethanolic extract of Citrullus colocynthis L. at 50, 100 & 200 mg/kg bw/day for 7 days (p.o). Group 6: The animals were treated with Silymarin (100 mg/kg p.o) which served as standard. Groups 3 to 6 were intoxicated with paracetamol (500 mg/kg bw) 1 h before the administration of extract or Silymarin for 10 days. Histopathological findings, different hepatic biochemical parameters viz. AST, ALT, ALP, Total bilirubin, Total cholesterol, Triglycerides, & the body weight before & after treatment were evaluated to investigate the hepatoprotective activity. Results: Paracetamol induced a significant rise in AST, ALT, ALP, Total Bilirubin, Total Cholesterol, Triglycerides. Administration of 200 mg/kg bw of ethanolic extract of Citrullus colocynthis L. effectively reduced these pathological damages caused by paracetamol intoxication. In addition to serum parameters treatment of 200 mg/kg bw of ethanolic extract of Citrulus colocynthis L. also promotes the body weight in albino rats as shown in Figure 6 respectively. Histopathological changes of the liver samples were compared with the normal control as shown in Figures 2-5 respectively. Conclusion: From our results we may infer that the mode of action of 90% ethanolic extract of Citrullus colocynthis L. (200 mg/kg bw) in affording the in-vivo hepatoprotective activity against paracetamol may be due to the cell membrane stabilization, hepatic cell regeneration & normalizing the serum parameters.

Phytochemical investigation, antioxidant and wound healing activities of Citrullus colocynthis (bitter apple)

Objective: To undertake metabolite profiling of various plant parts of Citrullus colocynthis, and assess antioxidant and wound healing activities of fractions for therapeutical applications. Methods: Extracts from leaves, stem, root, fruit pulp and seeds were analyzed using gas chromatography-mass spectrometry and high performance liquid chromatography. Variation in antioxidant potential was assayed by 1,1-diphenyl-2-picrylhydrazyl(DPPH) free radical scavenging assay. The extract with highest antioxidant potential was subjected on in-vivo wound healing activity using excision wound model. Results: Metabolite profiling of Citrullus colocynthis identified 70 chemically diverse metabolites from different plant parts by using a combination of GC-MS and HPLC. Concentration of colocynthin, a principal active secondary metabolite, ranged from 3.15 mg/g dry weight to 242.00 mg/g dry weight, the lowest being in leaves and highest in fruit pulp. DPPH radical scavenging activity of free radical(IC50) ranged from 196.44 μg/mL in fruit pulp to 413.33 μg/mL in leaves tissues. Significant wound contraction and increase in hydroxyproline content of granulation tissue were observed with ointment formulated from methanolic extract of fruit pulp. Conclusions: The study indicates that the methanol extract of Citrullus colocynthis fruit pulp when applied topically may promote wound contraction in rat model attributable to the accumulation of colocynthin. The high quantity of colocynthin(242.00 mg/g dry weight) and substantial concentration of 2,4-di-tert butyl phenol(3.2%), squalene(4.2%) and δ-tocopherol(2.5%) make this plant to provide new opportunities for development of medicinal, nutraceutical and dietary supplements with optimized functionality.

Preparation and Characterization of Alkyd Resins Based on Citrullus colocynthis Seed Oil

In this study,different samples of alkyd resins based on Citrullus colocynthis seed oil(CCSO)—which has a high content of linoleic acid—were prepared as a renewable raw material.Short(I),medium(II),and long(III)alkyd resins were synthesized using oil,glycerol,and phthalic anhydride(PA)in different proportions.Prunus mahaleb seed oil(PMSO),which contains conjugated linolenic acid,was blended with CCSO in different proportions to examine the effect of PMSO on the film properties of alkyd resins(IV and V).All synthesized resins were characterized by FTIR and 1H NMR spectroscopic analysis.These resins were also cured by an epoxy resin and their film properties(pencil hardness,adhesion,gloss,and chemical resistance)were determined.Analysis of these alkyds reveals that CCSO can be used as a new potential raw material for coating applications.

Colocynth (<i>Citrullus colocynthis</i>) Flesh Extract Suppresses Adipogenesis by Down-Regulating Adipogenic Transcription Factors and Their Target Genes in 3T3-L1 Preadipocytes

Citrullus colocynthis, a member of the Cucurbitaceae family, is widely distributed in North Africa. The fruits are recognized for their wide range of medicinal uses and promising pharmaceutical potential. The present study aimed to investigate the anti-obesity effect of the ethanol extract of colocynth flesh (FCEE) in 3T3-L1 cells following treatment at different doses. The viability of 3T3-L1 preadipocytes was analyzed via MTT assay and triglycerides were stained with Oil red O to assess lipid accumulation. Additionally, adipogenesis-related gene expression was quantified via qRT-PCR. FCEE (0 - 150 μg/mL) dose-dependently suppressed intracellular triglyceride accumulation during the adipogenesis by 23% and 66% at 100 and 150 μg/mL, respectively, but did not affect cell viability. Analysis of the time-dependence of the effect of FCEE demonstrated that the greatest anti-adipogenic activity was observed during the early stages of differentiation. FCEE also decreased GPDH activity in a dose-dependent manner, with 98% decrease observed at 150 μg/mL. In addition, at same range of FCEE concentrations, the main transcription factors, including CCAAT/enhancer binding protein α (C/EBPα), peroxisome proliferator activated receptor γ (PPARγ), and sterol regulatory element-binding protein 1c (SREBP-1c), were downregulated by 90%, 89%, and 89%, respectively at 150 μg/mL. As these are the master regulators of adipogenesis. The inhibition of their downstream target genes was also observed. Colocynth may be useful in the treatment of obesity owing to its powerful effects on fat, which result in changes to adipocyte differentiation and fat mobilization.

Synergistic Hepatoprotective Effect of Feronia fimonia L,, Citrullus colocynthis L,, and Tribulus terrestris L, against Paracetamol Induced Hepatotoxicity in Swiss Albino Rats

Paracetamol toxicity induced a significant rise in AST （aspartate transaminase）, ALT （alanine transaminase）, ALP （alkhaline phisphatase）, total bilirubin, total cholesterol, Triglycerides. Administration of 150 mg/kg b.w of synergistic ethanolic extracts of Feronia limonia L., Citrullus colocynthis L., and Tribulus terrestris L. effectively reduced these pathological damages caused by paracetamol intoxication. In addition to serum parameters treatment of 150 rng/kg b.w of synergistic ethanolic extract of Feronia limonia L., Citrullus colocynthis L. and Tribulus terrestris L. also promote the body weight in albino rats. Histopathological changes of the liver samples were compared with the normal control. From our results, we may infer that the mode of action of synergistic 90% ethanolic extracts of Feronia limonia L., Citrullus colocynthis L., and Tribulus terrestris L. （150 mg/kg b.w） in affording the in vivo hepatoprotective activity against paracetamol may be due to the cell membrane stabilization, hepatic cell regeneration and activation of antioxidative enzymes such as glutathione reductase, glutathione peroxidase, superoxide dismutase and catalase.

Effects of Citrullus colocynthis Seed Aqueous Extracts upon Sodium Transport across A6 Kidney Cell Monolayers

Background:Various extracts from Citrullus colocynthis seeds were recently examined for their possible favorable effects on glucose homeostasis,other metabolic variables,and pancreatic islet size in streptozotocin-induced diabetic rats.Objective:Considering the effectiveness of insulin upon target cells as another process susceptible to be influenced by plant extracts,the present study is used to compare the effects of either crude or defatted aqueous extracts from C.colocynthis seeds upon Na+transport in A6 kidney cells.Method:The intensity of electrical currents,taken as the potential difference divided by the epithelial resistance was measured in monolayers of A6 kidney cells exposed at the basolateral side to either crude untreated aqueous(EI)or defatted aqueous(EII)extracts,as well as their fractions EI1 and EII1 containing components with high molecular weight(>5 kDa),and EI2 and EII2 containing components with low molecular weight(<5 kDa).The effect of insulin was to assess the functional integrity of the A6 monolayer.Results:All extracts caused a rapid and sustained increase in current intensity.The defatted aqueous extract(EII)was more efficient than the non-defatted aqueous extract(EI)in terms of increasing the intensity of current in the A6 monolayer.The components of the defatted aqueous extract responsible for such a greater efficiency were located in the fraction containing components with low molecular weight and acted in a time-related progressive fashion upon Na+transport.Conclusion:These data suggest that colocynth seed extracts mainly containing components with low molecular weight increased the current intensity in A6 cells,thus suggesting that colocynth may affect Na+transport in kidney cells.

A New Flavone C-glycoside from Citrullus colocynthis

Objective To study the chemical constituents of Citrullus colocynthis. Methods The chemical constituents were isolated and purified by column chromatography on silica gel and Sephadex LH-20 and recrystallization as well. NMR spectra and physicochemical property were characterized for structural identification. Results Eleven compounds were isolated and identified as β-sitosterol (1), α-spinasterol-3-O-β-D-glucopyranoside (2), α-spinasterone (3), bis (2-ethylhexyl) phthalate (4), p-hydroxybenzoic acid (5), 6-C-p-methylbenzoylvitexin (6), dihydrocucurbitacin E (7), cucurbitacin E (8), dihydro-epi-iso-cucurbitacin D (9), dihydroisocucurbitacin B-25-acaetate (10), and cucurbitacin E 2-O-β-D-glucopyranoside (11). Conclusion Compound 6 is a novel compound. Compounds 1-5, 7, 9, and 10 are isolated from C. colocynthis for the first time.

Effects of Citrullus colocynthis L. in a rat model of diabetic neuropathy

Objective: This study investigated the biochemical, histopathological and physiological effects of Citrullus colocynthis on peripheral neuropathy in rats with streptozotocin(STZ)-induced diabetes.Methods: Seventy adult male Sprague-Dawley rats were included in the present study. Diabetes was induced in 60 rats, with a single intraperitoneal injection of STZ(65 mg/kg). After 4 weeks, the diabetic rats were assessed for neuropathy. Then, the diabetic rats with neuropathy were randomly divided into 6 groups for a 4-week treatment with gabapentin, oral administration of C. colocynthis fruit pulp powder(100 and 300 mg/kg per day), topical preparations as oil-based solution and ointment, or placebo.Changes in metabolic, physiological, biochemical and histological parameters were considered as treatment outcomes.Results: Metabolic outcomes(body weight and blood glucose level) were improved in the C. colocynthistreated groups as compared to placebo. Tail-flick and hot-plate tests also had lower latency in the C. colocynthis-treated groups. Measurement of oxidative stress markers(malondialdehyde, superoxide dismutase and catalase) showed the antioxidant effect of C. colocynthis. Histological evaluation of the sciatic nerve showed that C. colocynthis decreased the number of demyelinated and degenerated nerve fibers.Among the C. colocynthis-treated groups, the one receiving 100 mg/kg power per day orally had the best treatment outcomes.Conclusion: The present study showed that C. colocynthis fruit, through its antioxidant and hypoglycemic activities, has a positive effect in the treatment of diabetic neuropathy.

Protection of Citrullus colocynthis Fruit Extracts on Carbon Tetrachloride-induced and Bacillus Calmette-Guerin plus Lipopolysaccharide-induced Hepatotoxicity in Mice

Objective To investigate the hepatoprotective activities of the extracts from Citrullus colocynthis(ECC),a native plant used as traditional Uigur Medicine on acute liver injury in mice.Methods The activities of ECC of petroleum ether(ECCPE),chloroform(ECCC),ethyl acetate(ECCEA),n-butyl alcohol(ECCBA),and water(ECCW) were evaluated in vivo using two experimental models,carbon tetrachloride(CCl4)- and bacillus calmette-guerin(BCG) plus lipopolysaccharide(LPS)-induced acute hepatotoxicity in mice.The contents of aspartate aminotransferase(AST) and alanine aminotransferase(ALT) in serum were determined and the liver histological examination was carried out,respectively.Results The pretreatment with ECC for 7 d obviously reduced the impact of CCl4toxicity on the serum markers of liver damage,ECCEA and ECCC with a significant difference of AST(P < 0.01,0.05,respectively) and ALT(P < 0.05,0.01,respectively).The protective activity was reconfirmed against BCG + LPS-induced injury and the serum enzymatic levels were obviously elevated,for ECCEA and ECCC with a significant difference of AST(P < 0.05,0.01,respectively) and ALT(P < 0.01,0.05,respectively).Conclusion That ECCEA and ECCC are the potent hepatoprotective extracts that could protect liver against the acute injury,and this ability might be attributed to their hepatoprotective potentials.