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Title: Evaluation of pharmacological potentials and biological activities of Periploca aphylla Decne. and Fagonia olivieri DC.
Authors: Rashid, Umbreen
Keywords: Natural Sciences
Issue Date: 2015
Abstract: Medicinal plants provide rich resources of ingredients which can be used in drug development and synthesis. In Pakistan Periploca aphylla Decne. (Asclepiadoideae) is used to treat gastrointestinal, dermatological and topical diseases whereas Fagonia olivieri (Zygophyllaceae) is generally used for the cure of cancer, diabetes, asthma, fever, toothache, urinary discharges, stomach troubles and renal disorders. This study was designed to evaluate the pharmacological activities of P. aphylla and F. olivieri using in vitro and in vivo models. The fresh samples (whole plant) of P. aphylla and F. olivieri were collected and made in to a fine powder after shade drying. Methanol extract of powder was resolved with escalating polarity to get fractions; n-hexane, chloroform, ethyl acetate, n-butanol and residual aqueous fraction. The extract/fractions were subjected to phytochemical analysis which showed the existence of tannins, saponins, terpenoids, cardiac glycosides, alkaloids, phlobatannins and flavonoids. All the extracts/fractions showed potent activity against Gram-negative, Gram-positive bacteria and fungal species. However, the most active fractions of F. olivieri were ethyl acetate, n-butanol and aqueous while methanol, n-butanol and aqueous fractions of P. aphylla exhibited activity against a maximum number of strains as compared to other fractions. These fractions also exhibited good cytotoxic activity as the maximum (96.66 %) was shown by ethyl acetate fraction of the F. olivieri and methanol extract of P. aphylla at 1000 mg/ml. Total phenolic and flavonoid content of P. aphylla was found to be higher in the ethyl acetate (186±1.18 mg GAE/g of extract) and n-butanol fractions (85.1667±0.13 mg RE/g extract) respectively. The ethyl acetate fraction exhibited the lowest IC50 values for DPPH and ABTS radicals. The n-butanol fraction exhibited the radical scavenging activity for superoxide and OH radical and H2O2 scavenging. The total phenolic and flavonoid contents of the F. olivieri samples ranged from 16 ± 0.881 to 50 ± 1.764 mg GAE/g extract and 19 ± 0.529 to 106 ± 0.892 mg RTE/g extract, respectively. The antioxidant activity was measured by a number of in vitro systems. The aqueous extract showed the highest radical scavenging activity with IC50 = 55 ± 1.212 μg/ml for DPPH, followed for ABTS, H2O2, and superoxide radical. However, ethyl acetate fractions exhibited the lowest IC50 values for phosphomolybdenum and OH radical scavenging assays. xxiii Quantification of phenolic compounds was carried out using high performance liquid chromatography (HPLC) which showed the presence of rutin, catechin, myricetin and caffeic acid in the crude extract and fractions of P. aphylla and F. olivieri. Thirty compounds were identified in P. aphylla methanol extract during GC-MS analysis which revealed that hexadecanoic acid, methyl ester (13.65%) was a major component followed by 1,1,6-trimethyl-3-methylene-2-(3,6,9,13 tetramethyl-6- ethenye-10,14-dimethylene-pentadec-4-enyl)cyclohexane (9.41%), 9-Octadecenal, (Z)- (CAS) (8.00%), Pyrrolidine, 1-(6-phenyl-1-cyclohexen-1-yl)- (CAS) (7.50%) and 5 (Dimethylamino)-3-ethoxy-5-isopropyl-1-(4'-trifluoromethylphenyl)-2-[2"-(4- "'trifluoromethylphenyl) ethynyl]-1,3-cyclopentadiene (5.28%). The prevailing compounds in F. olivieri were methyl 1-(but-2'-en-1'-yl)-6,7- (dimethoxy)isoquinoline-3-carboxylAte (71.89%), ethyl hydrogen P,5'-anhydro-2',3'- O-isopropylideneadenosine-8-phosphonate (7.64%), 2-Bromo-5-(3,5-di-tert-butyl-4 (trimethylsiloxy)phenyl)furan (3.33%) and [1]Benzothieno[2',3'-3,4]thieno[3'',2''- 7,8]cycloocta[1,2-b:5,6-b']diquinoxaline (3.25%). Piscidic acid (C11H12O7) was isolated and identified by 1H-NMR, 13C-NMR and mass spectrum studies from aqueous fraction of F. olivieri, as an antibacterial compound isolated for the first time using activity guided fractionation. Anti-inflammatory and anti-depressant activities were measured by means of carrageenan induced paw edema and forced swim test (FST) in Sprague-Dawley rats respectively. The maximum activity was shown by n-butanol and methanol fractions of P. aphylla while ethyl acetate and aqueous fractions of F. olivieri in the above assays. The anti-diabetic potential of P. aphylla and F. olivieri was measured using glucose tolerance test and streptozotocin induced diabetes. The aqueous fraction of the F. olivieri and the n-butanol fraction of P. aphylla showed anti-diabetic potential. The effect of methanol extract of P. aphylla and F. olivieri was also investigated against paracetamol, gentamicin and doxorubicin-induced toxicity in Sprague-Dawley rats. At the end of the experiments, body weight and organ (liver, kidney, heart, lungs, testes, brain) weights were measured. Serum ALT, ALP, AST, total bilirubin, total protein, urea, creatinine and lipid profiles such as cholesterol, triglycerides, HDL and LDL were assayed. The levels of cardiac biomarker enzymes viz LDH, CK, CK-MB were observed. Lipid peroxidation (indexed by TBARS) and antioxidants like glutathione, glutathione transferase, peroxidase, superoxide dismutase and catalase xxiv were assessed. Histopathology of tissues was also performed. The genetic parameters were evaluated by means of DNA fragmentation and DNA ladder assay. The lesions induced by these drugs in all the above mentioned parameters were determined to be lower particularly in the P. aphylla and F. olivieri treated groups. Moreover, extracts administered at 400 mg/kg were found to show greater protective effects than that at 200 mg/kg. Furthermore, the protective activity of these extracts was comparable to that of silymarin, an active moiety in Silybum marianum, wellknown for its hepatic regenerative activity. Thus the results show that crude methanol extracts and partitioned fractions of P. aphylla and F. olivieri possess antimicrobial, cytotoxic, anti-inflammatory, antidepresssent, anti-diabetic and antioxidant effects. The spectra of activities displayed by the extracts could be credited to the presence of these phytochemicals (flavonoids, saponins, tannins, alkaloids and terpenoids). This may validate the use of these plants in traditional medicine for the treatment of various disease conditions and signifies the potential of P. aphylla and F. olivieri as sources of therapeutic agents.
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