Khan Usmanghani

Bactericidal activity of medicinal plants, employed for the treatment of gastrointestinal ailments, against Helicobacter pylori

AIM OF THE STUDY Helicobacter pylori infection plays a crucial role in the pathogenesis of peptic ulcer, and gastric cancer. The current PPI-based triple regimens for the eradication of Helicobacter pylori faces uprising resistance problem demanding for the search of novel candidates. Medicinal plants have always been a source of lead compounds for drug discovery. In the present study, we evaluated the anti-Helicobacter pylori activity of 50 commonly used Unani (traditional) medicine plants from Pakistan that are extensively utilized for the cure of gastrointestinal disorders to explore the natural source for pilot compounds against Helicobacter pylori. MATERIALS AND METHODS Total seven clinical isolates and one standard strain were employed to examine the bactericidal effects of medicinal plants. Helicobacter pylori was isolated from the antral biopsy specimens and confirmed through the standard microbiology procedures. Minimum bactericidal concentration (MBC) of the active plants was determined at the concentration range from 7.8 to 500 microg/ml. RESULTS Among the herbs evaluated, more than 50% inhibited the growth of eight strains at the concentration of 500 microg/ml. The 70% aqueous-ethanol extracts of Curcuma amada Roxb., Mallotus phillipinesis (Lam) Muell., Myrisctica fragrans Houtt., and Psoralea corylifolia L. demonstrated strong anti-Helicobacter pylori activity with MBC value ranged from 15.6 to 62.5 microg/ml. The most potent bactericidal activity was exhibited by Mallotus phillipinesis (Lam) Muell. which completely killed the bacteria at the concentration of 15.6-31.2 microg/ml. CONCLUSION The results revealed significant anti-Helicobacter pylori activity of medicinal plants which could be the potential source of new bactericidal agents.

Anti-inflammatory and cytoprotective effects of selected Pakistani medicinal plants in Helicobacter pylori-infected gastric epithelial cells.

ETHNOPHARMACOLOGICAL RELEVANCE Helicobacter pylori infection is associated with gastritis, peptic ulcer, and gastric cancer. Due to its high global prevalence and uprising resistance to available antibiotics, efforts are now directed to identify alternative source to treat and prevent associated disorders. In the present study, effect of selected indigenous medicinal plants of Pakistan was evaluated on the secretion of interleukin-8 (IL-8) and generation of reactive oxygen species (ROS) in a bid to rationalize their medicinal use and to examine the anti-inflammatory and cytoprotective effects in gastric epithelial cells. MATERIALS AND METHODS AGS cells and clinically isolated Helicobacter pylori strain (193C) were employed for co-culture experiments. Anti-Helicobacter pylori activity and cytotoxic effects of the selected plants were determined by serial dilution method and DNA fragmentation assay respectively. ELISA and flow cytometry were performed to evaluate the effect on IL-8 secretion and ROS generation in Helicobacter pylori-infected cells. RESULTS At 100μg/ml, extracts of Alpinia galangal, Cinnamomum cassia, Cinnamomum tamala, Mentha arvensis, Myrtus communis, Oligochaeta ramose, Polygonum bistorta, Rosa damascena, Ruta graveolens, Syzygium aromaticum, Tamarix dioica, and Terminalia chebula exhibited strong inhibitory activity against IL-8 secretion. Of these, four extracts of Cinnamomum cassia, Myrtus communis, Syzygium aromaticum, and Terminalia chebula markedly inhibited IL-8 secretion at both 50 and 100μg/ml. Cinnamomum cassia was further assessed at different concentrations against Helicobacter pylori and TNF-α stimulated IL-8 secretion, which displayed significant suppression of IL-8 in a concentration-dependent-manner. Among the plants examined against ROS generation, Achillea millefolium, Berberis aristata, Coriandrum sativum, Foeniculum vulgare, Matricaria chamomilla and Prunus domestica demonstrated significant suppression of ROS from Helicobacter pylori-infected cells (p<0.01). CONCLUSION Results of the study revealed anti-inflammatory and cytoprotective effects of selected medicinal plants which could partially validate the traditional use of these plants in GI disorders particularly associated with Helicobacter pylori. Furthermore, results obtained may lead to possible future candidates of chemoprevention against peptic ulcer or gastric cancer.

Modulation of Activation-Induced Cytidine Deaminase by Curcumin in Helicobacter pylori-Infected Gastric Epithelial Cells

Background:  Anomalous expression of activation‐induced cytidine deaminase (AID) in Helicobacter pylori‐infected gastric epithelial cells has been postulated as one of the key mechanisms in the development of gastric cancer. AID is overexpressed in the cells through nuclear factor (NF)‐κB activation by H. pylori and hence, inhibition of NF‐κB pathway can downregulate the expression of AID. Curcumin, a spice‐derived polyphenol, is known for its anti‐inflammatory activity via NF‐κB inhibition. Therefore, it was hypothesized that curcumin might suppress AID overexpression via NF‐κB inhibitory activity in H. pylori‐infected gastric epithelial cells.

Anti-inflammatory effect of cinnamaldehyde in Helicobacter pylori induced gastric inflammation.

Cinnamomum cassia is widely employed for gastrointestinal complaints such as dyspepsia, flatulence, diarrhea, and vomiting. Studies report cinnamaldehyde (CM) as a major active constituent of cinnamon. The aim of this study was to evaluate the anti-inflammatory mechanism of CM on Helicobacter (H.) pylori-infected gastric epithelial cells in order to validate cinnamon traditional use in gastrointestinal (GI)-related disorders. AGS/MKN-45 cells and H. pylori (193C) were employed for co-culture experiments. Anti-H. pylori cytotoxic and anti-adhesion activity of CM were determined. Enzyme linked immunosorbent assay, real time polymerase chain reaction analysis and immunoblotting were used to measure the effect on interleukin-8 (IL-8) secretion/expression. The effect on activation of nuclear factor kappa B (NF-κB) was determined by immunoblot analysis. The non-cytotoxic CM (≤125 µM) was also non-bactericidal at the given time, suggesting the effect in H. pylori/cell co-culture system was not due to alteration in H. pylori viability or the toxicity to the cells. Also, CM did not show any anti-adhesion effect against H. pylori/cell co-culture. However, pre-incubation of the cells with CM significantly inhibited the IL-8 secretion/expression from H. pylori-infected cells (p<0.01). In addition, CM suppressed H. pylori-induced NF-κB activation and prevented degradation of inhibitor (I)-κB This study provides evidence that the anti-inflammatory effect of C. cassia on H. pylori-infected gastric cells is due to blockage of the NF-κB pathway by cinnamaldehyde. This agent can be considered as a potential candidate for in vivo and clinical studies against various H. pylori related gastric pathogenic processes.

Kinetics of thermal degradation of betamethasone valerate and betamethasone dipropionate in different media

The effect of pH, media, phosphate concentration and ionic strength on the kinetics of thermal degradation of betamethasone valerate and betamethasone dipropionate has been investigated. A validated HPLC method has been used to determine the parent compounds and their major thermal degradation products identified in the reaction. Betamethasone-17-valerate gave rise to two major products, namely, betamethasone-21-valerate and betamethasone alcohol, and betamethasone dipropionate degraded into three major products, namely, betamethasone-17-propionate, betamethasone-21-propionate and betamethasone alcohol, in different media. Betamethasone valerate showed maximum stability at pH 4-5 while betamethasone dipropionate was maximally stable at pH 3.5-4.5. The degradation of betamethasone valerate and betamethasone dipropionate was found to follow first-order kinetics and the apparent first-order rate constants (kobs) for thermal degradation in different media range from 0.399-9.07×10-3 h-1 and 0.239-1.87×10-3 h-1, respectively. The values of the rate constants decrease with increasing solvent polarity, phosphate concentration and ionic strength. The second-order rate constants (k΄) for the phosphate ion inhibited reactions lie in the range of 3.02-1.30×10-6 M-1 s-1.

Clinical evaluation to assess the safety and efficacy of coded herbal medicine "Dysmo-off" versus allopathic medicine "Diclofenac sodium" for the treatment of primary dysmenorrhea.

The purpose of the present research work was to carry out clinical study on primary dysmenorrhea to comparatively examine the coded herbal drug formulation “Dysmo-off” with authentic allopathic medicine “Diclofenac sodium” (NSAIDs). A random controlled clinical trial was conducted to compare the efficacy and safety of coded herbal medicinal treatments Dysmo-off with Diclofenac sodium/Phenylacetic acid. These evaluations were based on verbal rating scale so as to ascertain the rate of analgesic effects on dysmenorrhoeic pain. The patients were randomly allocated with the ratio of 1:2 for controlled treatment with nonsteroidal anti-inflammatory drugs (NSAIDs) (n = 40) received Diclofenac sodium tablets twice daily for 4 days (50 mg one day prior to and three days after the menstruation), and test treatment with Dysmo-off (n = 80) received powdered Dysmo-off twice daily for four days (5 g one day prior to and three days after the menstruation). Treatment lasted for 4 consecutive menstrual cycles. Hemoglobin, ESR and ultrasound were measured at baseline during study. All subjects were clinically studied and completed the assigned therapy during the period May 2001 to June 2004.

In vitro evaluation of betamethasone esters for phototoxic potential.

The phototoxicity of Betamethasone valerate and betamethasone dipropionate has been investigated on irradiation in the wavelength range of 300–400 nm, using some basic in vitro phototoxicity tests. Both esters cause photohemolysis of mouse red blood cells and photoperoxidation of linoleic acid. The photoproducts of both esters have also been found to be phototoxic. The toxicity of these photoproducts increases with further irradiation; however, they exhibit toxicity, even in the dark. The efficient inhibition of photohemolysis by the well-known free radical scavengers, such as butylhydroxyanisole and reduced glutathione, and minor inhibition by singlet-oxygen quenchers, such as sodium azide, suggests that cellular damage is mainly mediated by free radicals whereas singlet oxygen plays a minor role.

Quantitative Determination of Catechin as Chemical Marker in Pediatric Polyherbal Syrup by HPLC/DAD

Vivabon syrup is a balanced composition of dietary ingredients of phytopharmaceutical nature for maintaining the physique, vigor, vitality and balanced growth of children. The herbal ingredients of pediatric syrup are rich in bioflavonoid, proteins, vitamins, glycosides and trace elements. Vivabon is formulated with herbal drugs such as Phoenix sylvestris, Emblica officinalis, Withania somnifera, Centella asiatica, Amomum subulatum, Zingiber officinalis, Trigonella foenum-graecum, Centaurea behen and Piper longum Catechins are flavan-3-ols that are found widely in the medicinal herbs and are utilized for anti-inflammatory, cardio protective, hepato-protective, neural protection and other biological activities. In general, the dietary intake of flavonoids has been regarded traditionally as beneficial for body growth. Standardization of Vivabon syrup dosage form using HPLC/DAD has been developed for quantitative estimation of Catechin as a chemical marker. The method was validated as per ICH guidelines. Validation studies demonstrated that the developed HPLC method is quite distinct, reproducible as well as quick and fast. The relatively high recovery and low comparable standard deviation confirm the suitability of the developed method for the determination of Catechin in syrup.

Standardization of Biomarkers Gallic Acid and Berberine in Polyherbal Formulation Entoban Capsules by High-Performance Thin-Layer Chromatography—Densitometry

The technological improvement in the structural elucidation of natural compounds has made it probable to generate appropriate strategies for the analysis and standardization of plant-based medicines. An appliance of highly oriented hyphenated techniques provides a definite tool for herbal investigations. Therefore, the present study was directed towards the standardization of biomarkers gallic acid and berberine in polyherbal formulation Entoban capsules to ensure the quality of the herbal drugs. A rapid, simple, accurate, and specific high-performance thin-layer chromatography (HPTLC) method for the quantitative estimation of biomarkers berberine and gallic acid has been developed. HPTLC was performed to evaluate the presence of gallic acid and berberine applying toulene—ethyl acetate—formic acid—methanol (12:9:4:0.5 v/v) and ethanol—water—formic acid (90:9:1 v/v), as the mobile phase, respectively. The RF values (0.58 for gallic acid and 0.76 for berberine) in both sample and reference standard were found comparable under ultraviolet (UV) light at 273 nm and 366 nm, respectively. The method developed resulted in good-quality peak shape and enabled high-quality resolution of biomarkers. The present standardization undertaken reveals compliance with the analytical procedure; therefore, it is concluded that Entoban capsule is a well-standardized product. Standardization falls under the specific guidelines of quality herbal medicine following the prerequisite for global harmonization.

Monograph of Tribulus terrestris

Tribulus terrestris has long been used as a tonic and aphrodisiac, and a diuretic in Unani system of medicine. The diuretic effect was attributed to the presence of potassium salts in high concentration. So many studies have been done on pharmacological activities of T. terrestris. The major constituents of these plants are steroidal saponins namely: terrestrosins A, B, C, D and E, desgalactotigonis, Fgitonis, desglucolanatigoneis, gitnin etc. The biological activity exhibited by saponins include: pisicidal, antimicrobial, molluscicidal, haemolytic, antiviral, cytotoxic, antihepatotoxic, spermicidal, insecticidal, antioedematous, antiulcer analgesic, immunomodulatory, and sedative effects.