Asian Pacific Journal of Tropical Biomedicine

Issue 6,2020 Table of Contents

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  • 1  Enzyme-treated date plum leave extract ameliorates atopic dermatitis-like skin lesion in hairless mice
    Byoung Ok Cho Jae Young Shin Ji-Su Kim Denis Nchang Che Hyun Ju Kang Hyeon Jin Kang Hyeonhwa Oh Young-Soo Kim Seon Il Jang
    2020(6):239-247. DOI: 10.4103/2221-1691.283937
    [Abstract](31) [HTML](0) [PDF 1.09 M](210)
    Objective: To evaluate the effect of different extracts of Diospyros lotus leaves in atopic dermatitis. Methods: Diospyros lotus leaves were extracted in ethanol and treated with or without hydrochloric acid or α-rhamnosidase to obtain three different extracts-ethanol, acid-hydrolyzed, and enzyme-hydrolyzed leaf extracts of date plum. The myricitrin content in all samples was measured using HPLC analysis. In vitro antioxidant and anti-inflammatory activities of the extracts were determined by measuring DPPH radical scavenging activities and nitric oxide production in RAW264.7 cells, respectively. Sevenweek- old male hairless mice were used to evaluate the anti-atopic dermatitis effects of three extracts in vivo. Splenocytes and mast cells were used to further determine the anti-atopic dermatitis effects of the major compound in the ethanol leaf extract. Results: Enzyme-hydrolyzed leaf extract showed significant in vitro antioxidant and anti-inflammatory activities, and attenuated atopic dermatitis-like skin symptoms and clinical signs more significantly than ethanol and acid-hydrolyzed leaf extracts in 1-fluoro-2,4- dinitrobenzene and house dust mite antigen-treated hairless mice. Enzyme-hydrolyzed leaf extract also suppressed the serum level of immunoglobulin E, interleukin (IL)-4, tumor necrosis factor (TNF)-α, interferon (IFN)-γ, thymic stromal lymphopoietin, and thymus and activation-regulated chemokine in mice with atopic dermatitis. Furthermore, histological analysis revealed that enzymehydrolyzed leaf extract suppressed the increased epidermal thickness, dermal infiltration of inflammatory cells, and infiltration and degranulation of mast cells more markedly than the other two extracts in atopic dermatitis-like skin lesions. In addition, this extract effectively inhibited the production of IFN-α, IL-4, and thymus and activation-regulated chemokine compared with the other two extracts in concanavalin A-stimulated splenocytes. Myricitrin, a major compound of enzyme-hydrolyzed leaf extract, suppressed atopic dermatitis biomarkers in stimulated mouse splenocytes and HMC-1 human mast cells. Conclusions: These results suggest that enzyme-hydrolyzed leaf extract might be a potential candidate to treat atopic dermatitis.
    2  Leishmania tropica: The comparison of two frequently-used methods of parasite load assay in vaccinated mice
    Fatemeh Nemati Zargaran Mosayeb Rostamian Alisha Akya Hamid M. Niknam
    2020(6):248-253. DOI: 10.4103/2221-1691.283938
    [Abstract](54) [HTML](0) [PDF 838.04 K](195)
    Objective: To compare limiting dilution assay and real-time PCR methods in Leishmania tropica parasite load measurement in vaccinated mice. Methods: BALB/c mice were vaccinated by Leishmania tropica soluble Leishmania antigen or recombinant Leishmania tropica stressinducible protein-1 with/without adjuvant. After three vaccinations, mice were challenged by Leishmania tropica promastigotes. Two months after challenge, the draining lymph nodes of mice footpad were removed and parasite load was assayed by limiting dilution assay and real-time PCR methods. Limiting dilution assay was done by diluting tissue samples to extinction in a biphasic medium. For real-time PCR, DNA of the lymph nodes was extracted, equal dilutions of each sample were prepared and hot-start real-time PCR was done using appropriate primers. The data of the two methods were compared by appropriate statistical methods. Results: Both methods were able to measure different levels of parasite load in vaccinated/unvaccinated mice. In addition, wherever parasite load of a group was estimated high (or low) by one method, the estimated parasite load by another method was the same, although statistically significant differences were found between some groups. Conclusions: Both methods lead to approximately similar results in terms of differentiating parasite load of the experimental groups. However, due to the lower errors and faster process, the real-time PCR method is preferred.
    3  Formononetin alleviates diabetic cardiomyopathy by inhibiting oxidative stress and upregulating SIRT1 in rats
    Manisha J. Oza Yogesh A. Kulkarni
    2020(6):254-262. DOI: 10.4103/2221-1691.283939
    [Abstract](32) [HTML](0) [PDF 3.04 M](317)
    Objective: To evaluate the effect of formononetin on type 2 diabetic cardiomyopathy. Methods: Diabetes was induced by feeding high-fat diet for 2 weeks and administration of 35 mg/kg of streptozotocin in rats. Formononetin was administered at 10, 20 and 40 mg/kg for 16 weeks once a day. Plasma glucose, lipid parameters, and cardiac markers in blood samples were measured. Body weight and relative heart weight were recorded. Hemodynamic parameters, oxidative stress parameters and silence information regulator 1 (SIRT1) expression in cardiac tissue were estimated. Histopathological changes in cardiac tissue were also observed. Results: Formononetin significantly reduced the levels of glucose, triglycerides, cholesterol, low density lipoprotein, creatine kinase- MB, lactate dehydrogenase and aspartate aminotransferase. In addition, formononetin significantly improved hemodynamic parameters, alleviated oxidative stress and increased SIRT1 expression. Conclusions: The study indicates that formononetin can improve hyperglycemia and hyperlipemia, reduce oxidative stress and increase SIRT1 expression. It can be a potential therapeutic agent for diabetic cardiomyopathy.
    4  Moringa oleifera leaf ethanol extract ameliorates lead-induced hepato-nephrotoxicity in rabbits
    Nancy B. Mohamed Amira H. Mohamed Nashwa A. Abu-Aita Soad M. Nasr Somia A. Nassar Kawkab A. Ahmed
    2020(6):263-272. DOI: 10.4103/2221-1691.283940
    [Abstract](31) [HTML](0) [PDF 1.52 M](194)
    Objective: To evaluate the effect of Moringa oleifera leaf ethanol extract as an adjunct treatment on lead acetate induced hepatonephrotoxicity in rabbits. Methods: Thirty-six male New Zealand White rabbits were assigned into two main groups. The first group (14 rabbits) served as normal control. The second group (22 rabbits) was administered orally with lead acetate at a dose of 40 mg/kg/day, 5 days/week for 8 weeks. At the 4th and the 8th week of treatment, 6 animals (3 animals at each period) of the second group were sacrificed while the remaining animals (16 rabbits) were assigned randomly into 2 subgroups (8 rabbits each): treated and non-treated. The first subgroup was orally given 1 mL phosphate-buffered saline for further 4 weeks while the second subgroup was administered orally with Moringa oleifera leaf ethanol extract at a dose of 400 mg/kg/day for the same period. Blood samples were collected to determine hematological and serum biochemical indices. Tissue specimens were collected from the liver and kidney for evaluation of the oxidant/antioxidant markers and for histopathological examinations. Results: Lead acetate exposure decreased the mean body weight gain, hematocrit, hemoglobin, mean corpuscular volume, and lymphocytes count. Moreover, it markedly increased counts of monocytes and platelets, serum enzyme activity, levels of creatinine, total cholesterol, triglycerides, and low-density lipoprotein cholesterol. Malondialdehyde level was markedly increased while the reduced glutathione content was significantly decreased in liver tissue of lead intoxicated-rabbits. Histopathological alterations were also noticed in the liver and kidney of lead intoxicated rabbits. Moringa oleifera leaf ethanol extract significantly improved hematological and serum biochemical parameters and histopathological structure of the liver and kidney. Conclusions: Moringa oleifera leaf ethanol extract ameliorates hemato-biochemical and histopathological alterations caused by lead acetate and improves hepatic and renal functions.
    5  Standardized extract of Centella asiatica ECa 233 inhibits lipopolysaccharide-induced cytokine release in skin keratinocytes by suppressing ERK1/2 pathways
    Furoida Moolsap Supita Tanasawet Mayuree H. Tantisira Pilaiwanwadee Hutamekalin Varomyalin Tipmanee Wanida Sukketsiri
    2020(6):273-280. DOI: 10.4103/2221-1691.283941
    [Abstract](72) [HTML](0) [PDF 1.33 M](299)
    Objective: To evaluate the effect of standardized extract of Centella asiatica ECa 233 on inflammatory mediator production through cyclooxygenase-2 (COX-2), extracellular signal-regulated kinase 1/2 (ERK1/2) and nuclear factor-κB (NF-κB) pathway in keratinocyte HaCaT cells. Methods: HaCaT cells were treated with 0.1, 1, 10 and 100 μg/mL ECa 233 in the presence of lipopolysaccharide (LPS). Proinflammatory cytokines and prostaglandin E2 were assessed with ELISA. Western blotting was used to determine the inhibition of COX-2, ERK1/2 and NF-κB protein expression. Results: ECa 233 suppressed LPS-induced release of interleukin- 1β, tumor necrosis factor-α, and prostaglandin E2. ECa 233 also inhibited COX-2, phosphorylation of ERK1/2 and the activation of NF-κB. Moreover, the formation of reactive oxygen species (ROS) was decreased in response to LPS-inflamed keratinocytes. Conclusions: ECa 233 inhibits LPS-stimulated production of inflammatory mediators in keratinocytes via suppressing ERK1/2 and NF-κB pathways. The suppressive effect of ECa 233 may be related to an inhibition of ROS production.
    6  Response surface methodology-based optimization of ultrasound-assisted extraction of β-sitosterol and lupeol from Astragalus atropilosus (roots) and validation by HPTLC method
    Perwez Alam Nasir A. Siddiqui Ali S. Alqahtani Anzarul Haque Omer A. Basudan Saleh I. Alqasoumi Abdullah A. AL-Mishari M.U. Khan
    2020(6):281-292. DOI: 10.4103/2221-1691.283942
    [Abstract](42) [HTML](0) [PDF 1.29 M](223)
    Objective: To optimize the ultrasonication method for efficient extraction of β-sitosterol and lupeol from the roots of Astragalus atropilosus using Box-Behnken design of response surface methodology (RSM), and its validation by high performance thin layer chromatography (HPTLC) method. Methods: Ultrasonication method was used to extract β-sitosterol and lupeol from Astragalus atropilosus (roots). RSM was used to optimize the different extraction parameters viz. liquid to solid ratio (10–14 mL/g), temperature (60-80 ℃) and time (40–60 min) to maximize the yield of β-sitosterol and lupeol. The quantitative estimation of β-sitosterol and lupeol was done in chloroform extract of Astragalus atropilosus by validated HPTLC method on 10 cm×20 cm glass-backed silica gel 60F254 plate using hexane and ethyl acetate (8:2, v/v) as mobile phase. Results: A quadratic polynomial model was found to be most appropriate with regard to R1 (yield of total extraction; R2/% CV = 0.994 8/0.28), R2 (β-sitosterol yield; R2/% CV = 0.992 3/0.39) and R3 (lupeol yield; R2/% CV = 0.994 2/0.97). The values of adjusted R2/predicted R2/signal to noise ratio for R1, R2, and R3 were 0.978 2/0.955 1/48.77, 0.990 4/0.911 0/31.33, and 0.992 7/0.940 1/36.08, respectively, indicating a high degree of correlation and adequate signal. The linear correlation plot between the predicted and experimental values for R1, R2, and R3 showed high values of R2 ranging from 0.990 5-0.997 3. β-sitosterol and lupeol in chloroform extract of Astragalus atropilosus were detected at Rf values of 0.22 and 0.34, respectively, at λ max = 518 nm. The optimized ultrasonic extraction produced 8.462% w/w of R1, 0.451% w/w of R2 and 0.172% w/w of R3 at 13.5 mL/g liquid to solid ratio, 78 ℃ of temperature and 60 min of time. Conclusions: The experimental findings of RSM optimized extraction and HPTLC analysis can be further applied for the efficient extraction of β-sitosterol and lupeol in other species of Astragalus.

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