Karim Raafat

Amelioration of Diabetes and Painful Diabetic Neuropathy by Punica granatum L. Extract and Its Spray Dried Biopolymeric Dispersions

Aims. To evaluate the effect of Punica granatum (Pg) rind extract and its spray dried biopolymeric dispersions with casein (F1) or chitosan (F2) against Diabetes mellitus (DM) and diabetic neuropathy (DN). Methods. We measured the acute (6 h) and subacute (8 days) effect of various doses of Pg, F1, and F2 and the active compounds on alloxan-induced DM mouse model. We evaluated DN utilizing latency tests for longer period of time (8 weeks). In addition, the in vivo antioxidant activity was assessed utilizing serum catalase level. Results. The results proved that the highest dose levels of Pg extract, F1, F2 exerted remarkable hypoglycemic activity with 48, 52, and 40% drop in the mice glucose levels after 6 hours, respectively. The tested compounds also improved peripheral nerve function as observed from the latency tests. Bioguided fractionation suggested that gallic acid (GA) was Pg main active ingredient responsible for its actions. Conclusion. Pg extract, F1, F2, and GA could be considered as a new therapeutic potential for the amelioration of diabetic neuropathic pain and the observed in vivo antioxidant potential may be involved in its antinociceptive effect. It is highly significant to pay attention to Pg and GA for amelioration and control of DM and its complications.

Prunella vulgaris L. active components and their hypoglycemic and antinociceptive effects in alloxan-induced diabetic mice

Prunella vulgaris L. (Lamiaceae) (PV) is a herbaceous plant traditionally utilized in management of diabetes and it has immunomodulatory activity. In this study, acute and subchronic antidiabetic, in-vivo antioxidant and antinociceptive effects of PV were evaluated in alloxan-induced type 1 diabetes (T1D) in a mouse model. Bio-guided fractionation, isolation, RP-HPLC, and 1H and 13C NMR identification of the active components responsible for PV effects were determined. RP-HPLC analysis showed that PV contained rosmarinic acid (RA) 4.5%, caffeic acid (CA) 9.8% and p-coumaric acid (pCA) 11.6%. Bio-guided fractionation showed that PV most active fraction was rich in caffeic acid, hence named, caffeic acid-rich fraction (CARF). RP-HPLC, and 1H and 13C NMR experiments showed that CARF contained CA (93.4%) and RA (6.6%). CARF reduced blood glucose levels and improved in-vivo oxidative-stress. It also inhibited the carbohydrate-hydrolyzing enzymes (alpha-amylase and alpha-glucosidase) and reduced HbA1c levels more significantly (p≤0.05) than that of PV and equivalent amounts of CA or RA. For longer times, CARF had significantly (p≤0.05) increased serum-insulin, ameliorated thermal hyperalgesia and tactile allodynia more significantly (p≤0.05) than the effects of PV and equivalent amounts of CA or RA. Moreover, the tested compounds showed potential restoration of the lipid peroxide levels. Consequently, CARF and PV observed increase in serum-insulin, attenuation of alpha-amylase and alpha-glucosidase, and their antioxidant potentials might be responsible for their antidiabetogenic and antinociceptive properties. In conclusion, CARF isolated from PV could be a potential therapeutic agent to ameliorate T1D and related complications.

Anti-Inflammatory and Antioxidant Activities of Salvia fruticosa: An HPLC Determination of Phenolic Contents.

Objectives. Salvia fruticosa Mill. (S. fruticosa) is widely used in folk medicine. Accordingly, the present study was designed to evaluate the antioxidant and anti-inflammatory activities of S. fruticosa, and to determine the phenolic constituents of its extracts. Methods. The antioxidant activity was determined using 2,2-diphenylpicrylhydrazyl assay. Total phenolic contents were estimated using Folin-Ciocalteu reagent, and high-performance liquid chromatography was performed to identify phenolic constituents. To evaluate the anti-inflammatory activity, carrageenan-induced mouse paw edema was determined plethysmographically. Key Findings. Different plant extracts demonstrated strong radical scavenging activity, where the ethyl acetate extract had the highest value in the roots and the lowest in the aerial parts. This antioxidant activity was correlated to the total phenolic content of different extracts, where rutin and luteolin were the most abundant constituents. Interestingly, both the roots and aerial parts revealed a significant anti-inflammatory activity comparable to diclofenac. Conclusions. This study is the first to demonstrate pharmacologic evidence of the potential anti-inflammatory activity of S. fruticosa. This activity may partly be due to the radical scavenging effects of its polyphenolic contents. These findings warrant the popular use of the East Mediterranean sage and highlight the potential of its active constituents in the development of new anti-inflammatory drugs.

Hypoglycemic effects of Prunus cerasus L. pulp and seed extracts on Alloxan-Induced Diabetic Mice with histopathological evaluation

AIM OF THE STUDY The study was undertaken to investigate the acute and subchronic hypoglycemic effects of an ethyl acetate (EtAc) extract of red sour cherry (Prunus cerasus) pulps and seeds in alloxan-induced diabetic mice. MATERIALS AND METHODS The Prunus cerasus pulp (PPc) and seed (SPc) EtAc extract s were IP-administered to the mice in doses of 100, 150 and 200mg/kgbody weight (b.w.) for acute study (6h) and subchronic study (8days). The in vivo effects of (PPc) and (SPc) EtAc extracts were assessed through blood glucose levels (BGL), changes in body weight as well as antioxidant enzyme activity. At the end of the study, animals were sacrificed for obtaining pancreatic tissues samples to assess histopathological changes in pancreas of control and alloxan-induced mice. RESULTS Acute administration of either PPc or SPc for 6h at all tested doses, significantly (P≤0.05) decreased BGL as compared with vehicle treated diabetic control (DIA CTRL) group with PPc showing more significant acute antidiabetic activity than those of SPc. In addition, subchronic administration of PPc has shown more significant (P≤0.05) amelioration of hyperglycemia, weight-loss and oxidative stress at all doses compared to SPc administration. Histologically, treatment with PPc and SPc showed significant pancreatic cell regeneration compared to diabetic controls. CONCLUSIONS Results indicated that ethyl acetate extracts of Prunus cerasus pulps and seeds promote acute and subchronic hypoglycaemic effects in alloxan-induced diabetic mice with pulp extracts having a more pronounced effect.

Phytotherapeutic activity of curcumol: Isolation, GC–MS identification, and assessing potentials against acute and subchronic hyperglycemia, tactile allodynia, and hyperalgesia

Abstract Context: Curcumol has recently attracted special attention due to its potential activities in many chronic disorders. Moreover, the traditional role of turmeric [Curcuma longa L. (Zingiberaceae)] in suppression of hyperglycemia is of great interest. Objectives: The present work explores the potential acute and subchronic antihyperglycemic, antinociceptive, and in vivo antioxidant effects of curcumol in alloxan-diabetic mice. Materials and methods: Bio-guided fractionation, column-chromatography, and GC–MS were utilized to identify the most active compound of turmeric (curcumol). Turmeric (25, 50, and 100 mg/kg), the curcumol rich fraction (CRF) (7 mg/kg), and curcumol (20, 30, and 40 mg/kg) were assessed for their acute (6 h) and subchronic (8 d) antihyperglycemic potentials and antinociceptive effects (8 weeks) were measured, using hot-plate and tail-flick latencies and von-Frey filaments method and in vivo antioxidant effects in alloxan-diabetic mice. Results: The most-active turmeric fraction was found to be rich in curcumol (45.5%) using GC–MS analysis method. The results proved that the highest dose levels of turmeric extract and curcumol exerted remarkable hypoglycemic activity with 41.4 and 39.3% drop in the mice glucose levels after 6 h, respectively. Curcumol (40 mg/kg) was found to be 9.4% more potent than turmeric extract (100 mg/kg) in subchronic management of diabetes. Curcumol also showed a significant improvement of peripheral nerve function as observed from the latency and tactile tests. Discussion: The antioxidant potential of curcumol may cause its ability to ameliorate diabetes and diabetes-related complications. Conclusions: Curcumol, a natural metabolite with a good safety-profile, showed results comparable with tramadol in reversing diabetes-induced tactile allodynia and hyperalgesia.

Infrared-Assisted Extraction and HPLC-Analysis of Prunus armeniaca L. Pomace and Detoxified-Kernel and their Antidiabetic Effects: Infrared-Assisted Phytochemical Analysis of Prunus armeniaca

INTRODUCTION Prunus armeniaca L. (P. armeniaca) is one of the medicinal plants with a high safety-profile. OBJECTIVES The aim of this work was to make an infrared-assisted extraction (IR-AE) of P. armeniaca fruit (pomace) and kernel, and analyse them using reverse phase high-performance liquid chromatography (RP-HPLC) aided method. METHODS IR-AE is a novel-technique aimed at increasing the extraction-efficiency. The antidiabetic-potentials of the P. armeniaca pomace (AP) and the detoxified P. armeniaca kernel extract (DKAP) were monitored exploring their possible hypoglycemic-mechanisms. Acute (6 h), subchronic (8 days) and long-term (8 weeks) assessment of Diabetes mellitus (DM) using glucometers and glycated hemoglobin (HbA1c) methods were applied. RESULTS Serum-insulin levels, the inhibitory effects on alpha-glucosidase, serum-catalase (CAT) and lipid peroxidation (LPO) levels were also monitored. AP was shown to be rich in polyphenolics like trans-lutein (14.1%), trans-zeaxanthin (10.5%), trans-ß-cryptoxanthin (11.6%), 13, cis-ß-carotene (6.5%), trans 9, cis-ß-carotene (18.4%), and ß-carotene (21.5%). Prunus armeniaca kernel extract before detoxification (KAP) was found to be rich in amygdaline (16.1%), which caused a high mortality rate (50.1%), while after detoxification (amygdaline, 1.4%) a lower mortality rate (9.1%) was found. AP showed significant (p ≤ 0.05, n = 7/group) antidiabetic-activity more prominent than DKAP acutely, subchronically and on longer-terms. IR-AEs displayed more efficient acute and subchronic blood glucose level (BGL) reduction than a conventional extraction method, which might be attributed to IR-AE superiority in extraction of active ingredients. AP showed more-significant and dose-dependent increase in serum-insulin, CAT-levels and body-weights more prominent than those of DKAP. Alpha-glucosidase and LPO levels were inhibited with AP-groups more-significantly. CONCLUSION In comparison to conventional-methods, IR-AE appeared to be an efficient and time-conserving novel extraction method. The antidiabetic-potentials of pomace and detoxified-kernels of P. armeniaca were probably mediated via the attenuation of glucose-provoked oxidative-stress, the inhibition of alpha-glucosidase and the marked insulin-secretagogue effect. Copyright

Phytochemical Compositions and Antidiabetic Potentials of Salvia sclarea L. Essential Oils

Salvia sclarea (SS) is characterized by its valuable essential oils (Eos) and potent biological activities. This study aimed at investigating the phytochemical composition of SS Eos collected in within the same week, from two different regions in Lebanon, Beirut (SS-Bt) and Taanayel (SS-Tl), utilizing GC-MS methods, and to explore their acute and subchronic antidiabetic potentials. Moreover, studying the phytochemical diversity of twenty SS Eos established on our work and literature descriptions in order to recognize the origin of the Lebanese active chemotype(s). The Eos have been obtained by hydro-distillation and identified via GC-MS analyses. Five chemotypes of SS Eos have been identified. The Lebanese Eos, SS-Bt and SS-Tl, studied here have shown evidence to belong to two different chemotypes 1 and 5, respectively. SS-Bt has shown to belong to chemotype 1, which is characterized by high linalool (LL) concentration (average 40.2%). On the other hand, SS-Tl has shown to belong to chemotype 5, which is characterized by high linalyl acetate (LA) concentration (average 50.4%). The acute and subchronic antidiabetic activities of these EOs have been monitored along with LL and LA, in order to find the most active chemotype. Chemotypes 1 (owned to high LL content), present at low altitude places of Lebanon and Poland, has shown significantly higher acute and subchronic antidiabetic activities than that of chemotype 5 (owned to high LA content). In conclusion, Salvia sclarea Eos have shown potential antidiabetic activities, and their Eos might be used in the future as a complementary or an alternative medicine in the management of diabetes and related complications.

Phytochemical Analysis of Nigella sativa L. Utilizing GC-MS Exploring its Antimicrobial Effects against Multidrug-Resistant Bacteria

Introduction: The alarming increase in bacterial strains resistant to existing antimicrobial agents has demanded alternative strategies. Medicinal plants are now considered as an alternative treatment because of their secure choice in several diseases. Among them, Nigella sativa is a promising traditional herb having rich medical background. The aim of the study is to perform phytochemical analysis of Nigella sativa L. Utilizing Gas chromatographic–mass spectrometric (GC/MS) exploring its antioxidant and antibacterial activity against multidrug resistant (MDR) gram-positive and gram-negative bacteria. Methods: Total phenolic, tannin, and flavonoid content of N. sativa seed extracts and its commercially available oil were determined. Their radical scavenging activity using DPPH was also tested. The antibacterial activity of N. sativa seed extracts and its oil against MDR gram-positive and gram-negative bacterial strains was studied using disc diffusion test and the biofilm formation assay. GC-MS studies were also performed. Results: Among the different preparations used, N. sativa oil showed the highest antioxidant and antibacterial activity against highly resistant gram-positive bacteria with the greatest suppression of biofilm formation, which was attributed to its high bioactive contents. Conclusion: This study indicates that N. sativa extracts and its oils can be used as natural antibacterial agents to treat infections caused by multidrug resistant bacteria.

Neuroprotective effects of Moringa oleifera: Bio-guided GC-MS identification of active compounds in diabetic neuropathic pain model

ObjectivesTo explore the phytotherapeutic-activities of Moringa oleifera (MO) seeds on painful diabetic neuropathy in alloxan-induced diabetic mice.MethodsThe bio-guided fractionation of MO utilizing column chromatography aided with GC-MS was used to detect the most active constituent of MO. Hyperalgesia, using tail-flick and hot-plate latency experiments, and mechanical-allodynia, utilizing von-Frey filaments, were evaluated before and after 8 weeks of intraperitoneal alloxan administration (180 mg/kg). Serum catalase and insulin levels, body weight and blood glucose levels (BGL), alpha-glucosidase inhibition, lipid peroxidation and glycated hemoglobin (HbA1c) were measured to evaluate both alloxan-induced diabetes mellitus and diabetic painful neuropathy (DPN).ResultsBeta-sitosterol (BSL) was proved to be the most active constituent of MO. The administration of MO (40, 60 and 80 mg/kg) or BSL (18, 25 and 35 mg/kg) significantly attenuated hyperalgesia and tactile allodynia (P⩽0.05), compared with tramadol (10 mg/kg) acting as a positive control, in alloxan-treated animals (n=7 per group). Moreover, MO and BSL have improved insulin secretion, in vivo antioxidant catalase, lipid peroxidation, acute and subchronic BGL, and normalized alpha-glucosidase and HbA1c levels.ConclusionsThe observed insulin secretagogue, alpha-glucosidase inhibition, hypoglycemic and antioxidant potentials might be responsible for MO and BSL antinociception and neuroprotective mechanism. MO and BSL have shown good glycemic-control and powerful neuroprotective properties which might serve as potential lead-compounds for further analysis.