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   Table of Contents - Current issue
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October-December 2020
Volume 11 | Issue 4
Page Nos. 157-237

Online since Saturday, October 10, 2020

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ORIGINAL ARTICLES  

The coronavirus disease 2019 main protease inhibitor from Andrographis paniculata (Burm.f) Ness Highly accessed article p. 157
Sukardiman , Martha Ervina, Mohammad Rizki Fadhil Pratama, Hadi Poerwono, Siswandono Siswodihardjo
DOI:10.4103/japtr.JAPTR_84_20  
The coronavirus disease 2019 (COVID-19) pandemic has attracted worldwide attention. Andrographis paniculata (Burm. f) Ness (AP) is naturally used to treat various diseases, including infectious diseases. Its Andrographolide has been clinically observed for anti-HIV and has also in silico tested for COVID-19 main protease inhibitors. Meanwhile, the AP phytochemicals content also provides insight into the molecular structures diversity for the bioactive discovery. This study aims to find COVID-19 main protease inhibitor from AP by the molecular docking method and determine the toxicity profile of the compounds. The results obtained two compounds consisting of flavonoid glycosides 5,4'-dihydroxy-7-O-β -D-pyran-glycuronate butyl ester and andrographolide glycoside 3-O-D-glucopyranosyl-andrographolide have lower free binding energy and highest similarity in types of interaction with amino acid residues compared to its co-crystal ligands (6LU7) and Indinavir or Remdesivir. The toxicity prediction of the compounds also reveals their safety. These results confirm the probability of using AP phytochemical compounds as COVID-19 main protease inhibitors, although further research must be carried out.
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Anticancer evaluation of N-benzoyl-3-allylthiourea as potential antibreast cancer agent through enhances HER-2 expression p. 163
Tri Widiandani, Siswandono , Edy Meiyanto
DOI:10.4103/japtr.JAPTR_77_20  
Breast cancer with HER-2 overexpression is sensitive to drugs which target the receptor or its kinase activity. Although the anti-HER-2 therapies commonly used have improved patient outcome, resistance usually occurs. In this present study, we investigated a modification of the chemical structure of allylthiourea derivatives in order to enhance the cytotoxicity effect on breast cancer cells with HER-2 overexpression. The aim of this research was to predict the absorption, distribution, metabolism, excretion, and toxicity by in silico study and to explore the effect N-benzoyl-3-allylthiourea (BATU) on MCF-7 cell line with overexpressing of HER-2 using MTT assay and western blot. The result showed that the cytotoxicity effects of BATU on MCF-7/HER-2 cell line (IC50value 0.64 mM) were higher than on MCF-7 cell lines (IC50value 1.47 mM). In addition, the cytotoxic effects of BATU on MCF-7 and MCF-7/HER-2 were higher than allylthiourea as a lead compound (IC50value 5.22 and 3.17 mM). The results also confirmed that the BATU compound has the ability to effectively enhance its cytotoxicity against MCF-7/HER-2 through enhanced HER-2 expression and inhibition of nuclear factor kappa B (NF-kB) activation. Above all, the BATU compound is effective in increasing HER-2 expression and inactivating NF-kB transcription factors, thereby resulting in inhibition of protein expression which works a significant part in cell proliferation. Therefore, the BATU compound has the potential to be developed as a complementary drug in breast cancer therapy with HER-2 positive.
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Effects of lime (Citrus aurantifolia) peel to the expression of mRNA toll-like receptors 4 in balb/c mice-infected Salmonella typhi p. 169
Vivien Novarina Kasim, Mochammad Hatta, Rosdiana Natzir, Veni Hadju, Ami Febriza, Hasta Handayani Idrus
DOI:10.4103/japtr.JAPTR_48_20  
Lime peel contains metabolic compounds that have lethal effects of bacterial cells, but its effect as an antibacterial modulate innate immunity pathways, especially toll-like receptor 4 (TLR-4) signaling pathway, is unclear. This study examined the effects of lime peel extract (LPE) on the activity of TLR 4 in Balb/c mice induced by Salmonella typhi. Mice were induced intraperitoneally and then 3 days after induction, LPE was given orally on two doses (510 and 750 mg/kg BW). The number of bacterial colonization was counted using peritoneal fluid samples by the method of plate count agar. Intervention LPE for 5 days can degrade TLR-4 and the number of colonies of S. typhi. On day 3 after was induced S. typhi, TLR-4 gene expression of Balb/c mice is increased. Postintervention LPE for 5 days, the expression of TLR-4 gene decreased, significantly different at a dose of 750 mg/kg BW (P = 0.04). There was a positive correlation between the expression of TLR-4 gene by the number of bacterial colonization, decreasing gene expression of TLR-4, the number of bacterial colonization is also getting smaller (P= 0.013, r = 0.408). LPE can modulate the TLR-4 signaling pathway in host immunity so that the gene TLR-4 is expressed fewer in numbers. This mechanism causes the bacterial colony number to decrease, not even growth.
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Anti-aging potential and phytochemicals of Centella asiatica, Nelumbo nucifera, and Hibiscus sabdariffa extracts p. 174
Monsicha Khuanekkaphan, Chanai Noysang, Warachate Khobjai
DOI:10.4103/japtr.JAPTR_79_20  
Centella asiatica, Nelumbo nucifera Gaertn, and Hibiscus sabdariffa have been used as medicinal plants in Thailand. They are sources of phytochemicals that applications for esthetic and healthcare. The aim of this research was to examine the phytochemical constituents and anti-aging potential of these plants. The phytochemical compounds were performed using gas chromatography-mass spectrometry. The anti-aging activities were evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azinobis (3-ethylbenzothiazoline-6-sunfonic acid) (ABTS), anti-collagenase, and anti-elastase assays. The main interest phytochemical compounds of ethanolic extracts of C. asiatica, N. nucifera, H. sabdariffa were ethanol, 2-(-Octadecenyloxy), γ-sitosterol and hexadecanoic acid, and ethyl ester, respectively. The DPPH half-maximal inhibitory concentration (IC50) results of C. asiatica, N. nucifera, and H. sabdariffa were 0.32 ± 0.01, 0.34 ± 0.00, and 0.35 ± 0.01 mg/mL, respectively. The ABTS result of H. sabdariffa extract showed high inhibitory activity at IC50of the extract was 0.62 ± 0.12 mg/mL. The percentage of collagenase inhibition of C. asiatica, N. nucifera, and H. sabdariffa at 1.0 mg/mL was 78.13 ± 4.42, 85.94 ± 2.21, and 90.63 ± 0.00, respectively. The C. asiatica extract had a high percentage of elastase inhibition. Consequently, these research results suggest that phytochemicals may also provide a range of esthetic and health benefits. The phytochemical constituent could be used as anti-aging active ingredient for cosmetic and pharmaceutical industrials.
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Proximate composition, vitamin and mineral composition, antioxidant capacity, and anticancer activity of Acanthopanax trifoliatus p. 179
Aunyachulee Ganogpichayagrai, Chutinan Suksaard
DOI:10.4103/japtr.JAPTR_61_20  
Acanthopanax trifoliatus has been used as both traditional plant food and medicinal plant in Thailand. This study aimed to evaluate proximate, vitamin, and mineral compositions of A. trifoliatus leaf samples together with antioxidant and anticancer activities of ethanolic leaf extract of A. trifoliatus. For leaf samples, quantitative determination of proximate composition was evaluated comprising moisture, crude protein, total fat, total carbohydrate, dietary fiber, ash, as well as energy. Quantitative determination of vitamin and mineral composition including Vitamin A, Vitamin B1, Vitamin B2, Vitamin C, calcium, sodium, and iron was also assessed. For ethanolic leaf extract, antioxidant activity was investigated using oxygen radical absorbance capacity (ORAC) and ferric reducing antioxidant power (FRAP) methods. Anticancer activity was determined against human ductal, bronchogenic, liver, gastric, and colon cancer cell lines using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. Proximate composition of A. trifoliatus was found to be 74.62 ± 0.38, 5.01 ± 0.05, 0.95 ± 0.04, 16.47 ± 0.40, 8.54 ± 0.06, and 2.95 ± 0.04 g/100 g sample, respectively, and energy was found to be 94.48 ± 1.30 kcal/100 g sample. Vitamin and mineral composition was found to be 428.47 ± 3.00 μg/100 g sample, 0.41 ± 0.01, 0.17 ± 0.00, 11.95 ± 0.86, 675.35 ± 46.57, 13.46 ± 0.95, and 4.79 ± 0.15 mg/100 g sample, respectively. A. trifoliatus ethanolic leaf extract revealed antioxidant capacity with ORAC value of 9057.29 ± 43.08 μmol TE/100 g and FRAP value of 1230.88 ± 19.51 μmol TE/100 g. Its extract also showed cytotoxic potential against all tested cancer cell lines. A. trifoliatus leaf is a good source of essential nutrients, which had antioxidant and anticancer potential.
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Stabilization of eye drops containing autologous serum and recombinant human epidermal growth factor for dry eye syndrome p. 184
Sriwidodo Sriwidodo, Insan Sunan Kurniawan Syah, Iman Permana Maksum, Toto Subroto, Ulvi Zasvia, Abd Kakhar Umar
DOI:10.4103/japtr.JAPTR_72_20  
Human epidermal growth factor (hEGF) and autologous serum are considered safer and more effective in treating dry eye syndrome. However, suitable formulas and preparation methods are needed to obtain eye drop containing autologous serum and hEGF, which are stable during storage and use. Therefore, this study aimed to develop a stable and effective eye drops containing autologous serum and hEGF. Stabilization of autologous serum and hEGF was done by adding lyoprotectant and antioxidant agents, and then prepared using the freeze-drying method. The clarity, pH, sterility, and endotoxin content of the preparation were evaluated. The effectiveness of the preparation was assessed by a cell viability test using a WST-8 reagent. Based on the results, all formulas produce preparations that are isotonic, clear, sterile, stable, and free from endotoxins. Cell viability test shows the addition of 25 μg/mL hEGF increased epithelial cell proliferation by up to 197%. It can be concluded that eye drops containing autologous serum and 25 μg/mL hEGF can be a promising therapy for dry eye syndrome.
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Antidiabetic activity of thin film containing astaxanthin-loaded nanoemulsion using carboxymethylcellulose sodium polymer on alloxan-induced diabetic rabbit p. 189
Lusi Nurdianti, Taofik Rusdiana, Iyan Sopyan
DOI:10.4103/japtr.JAPTR_55_20  
The present study was conducted to evaluate the potency of thin film containing astaxanthin-loaded nanoemulsion (FDT-As-NE) in lowering blood glucose levels on alloxan-induced diabetic rabbit (ADR). Astaxanthin nanoemulsion (As-NE) was prepared using self-nanoemulsifying method, followed by incorporated into the carboxymethylcellulose sodium matrix polymer using a solvent casting method to form a thin film. The evaluation of FDT-As-NE was performed by chemical, physical, and mechanical characterizations. The administration of thin film was done by an intraoral route. New Zealand albino rabbits were induced with alloxan to get experimental diabetic animals. The antidiabetic activity was carried out in three groups of treatment. Group I was ADR treated by FDT-As-NE, Group II was ADR treated by pure astaxanthin, while Group III was normal control. The measurement of fasting means blood glucose levels was carried out in 0 days (before treatment) and after 14 days of treatment. The histopathological analysis of the pancreas was also examined. Data were statistically evaluated using Kruskal–Wallis statistical test. P < 0.05 was considered statistically significant. FDT-As-NE had good physical and mechanical characteristics that suitable for intraoral administration. Group I reduced elevated blood glucose levels compared to Group II (P < 0.01). Histopathological examination of pancreatic tissue for a Group I showed the normal condition of pancreatic β-cell, suggesting the absence of any pathological lesions. These results suggest that thin film containing astaxanthin-loaded nanoemulsion administered by an intraoral route potentially useful for reducing glucose levels.
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In silico docking and comparative ADMET profile of different glycogen synthase kinase 3 beta inhibitors as the potential leads for the development of anti-Alzheimer drug therapy p. 194
Manisha Prajapat, Phulen Sarma, Nishant Shekhar, Hardeep Kaur, Sanjay Singh, Subodh Kumar, Harpinder Kaur, Saniya Mahendiratta, Amit Raj Sharma, Sukhmandeep Kaur, Vidya M Mahalmani, Bikash Medhi
DOI:10.4103/japtr.JAPTR_178_19  
Glycogen synthase kinase 3 beta (GSK3 β) plays a key role in pathologic hyper phosphorylation of tau and plays an important role in the pathogenesis of Alzheimer's disease. In the present study, we have screened a set of potential hits in in silico platform to gain insight regarding binding profile with the target (GSK3 β) from molecular docking, ADME/T, and molecular dynamics (MD) simulations. The three screened compounds 6-BIBEO, 6-BIO, and SB216763 topped the docking score chart when subjected to hard scoring function extraprecision of GLIDE. The active site dynamics study through MD simulations provides insights on residues Asp133, Val135, and Ile62 which are in a state of minimum deviation from their mean special position while they interact with the respective ligands. The same molecules also displayed favorable pharmacokinetic profile, negative Ames test and falls correctly within drug-likeliness rules. These agents can be taken forward further for the development of anti-Alzheimer's drug therapy.
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Effect of implantation Ti-6Al-4V ELI in femoral bone defect regeneration of Sprague Dawley rat p. 202
I Nyoman Jujur, Damisih , Kartika Devy, Shovy Suha, Boy Muchlis Bachtiar, Endang Winiati Bachtiar
DOI:10.4103/japtr.JAPTR_74_20  
Ti-6Al-4V ELI is one of the most commonly used dental implant restore function. The solution treatment temperature variation can significantly increase the strength, but it is not yet known the effect of these temperature variations on the alloy's biocompatibility properties. Twelve female Sprague Dawley rats were divided into six groups as follows: the treated group, the control group, and the defect group without implant material. In the treated group, the femur bone defect was implanted with as-cast Ti-6Al-4V ELI, 850°C, 950°C, and 1050°C heat-treated Ti-6Al-4V ELI implant material. The rats were euthanized after 30 days postimplantation and evaluated histologically. The results show that the histological scoring of the specimen for femur defect without implant material is 2 (fibrous union and fibrocartilaginous), score with implant as-cast is 2.5, the sample with 850°C heat treatment material is 2.5, 950°C is 2.5, and the temperature at 1050°C is 2.5. The score of 2.5 is between score 2 and score 3: hemorrhage, fibrous union, fibrocartilaginous microhemorrhage, and mineralized cartilage union. In conclusion, there is no effect of different heat treatment temperatures for Ti-6Al-4V ELI implant material in rat bone regeneration's maturation level.
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Development and validation of an ultra-performance liquid chromatography mass spectrometry/mass spectrometry method for simultaneous quantification of total and free mycophenolic acid and its metabolites in human plasma p. 207
Napatsanan Tanathitiphuwarat, Pannee Tanudechpong, Pajaree Chariyavilaskul, Nantaporn Prompila, Nat Tansrisawad, Apinya Tubtimrattana, Supeecha Wittayalertpanya
DOI:10.4103/japtr.JAPTR_40_20  
A reliable method has been validated using ultra-performance liquid chromatography mass spectrometry (MS)/MS for simultaneous evaluation of human plasma concentration of mycophenolic acid (MPA) and its major metabolites both total and free form. All analytes were extracted from plasma by simple protein precipitation procedure with methanol. Samples for determination of their free form concentration require a preanalytic spin through an ultrafiltration system. The chromatographic separation was completed using C18column at 0.3 ml/min with a gradient condition. Method validation was performed as the United State Food and Drug Administration guidelines for bio-analytical methods concerning precision, accuracy, linearity, selectivity, recovery, and matrix effect. Linearity was obtained over concentration of 0.05–4, 0.5–60, and 0.025–3 μg/ml for total MPA, mycophenolic acid glucuronide (MPAG) and mycophenolic acid acyl-glucuronide (AcMPAG), respectively. The linearity of the method for free form of analytes was confirmed in the range of 10–500, 125–10,000, and 0.5–300 ng/ml for MPA, MPAG, and AcMPAG, respectively. The intra- and interday accuracy ranged from 85.73%–102.01% for total form, and 87.23%–111.89% for free form, and the precisions of all analytes were lower than 15%. The mean recoveries of the analytes ranged from 85.54% to 94.76% and the matrix factor ranged from 0.88–1.06. The developed method is rapid, sensitive and convenient for pharmacokinetic study or therapeutic drug monitoring in patients after oral administration of enteric-coated mycophenolate sodium or mycophenolate mofetil.
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The enhancement apoptosis of osteosarcoma mesenchymal stem cells co-cultivation with peripheral blood mononuclear cells sensitized by secretome and granulocyte macrophage colony-stimulating factor p. 213
Ferdiansyah Mahyudin, Hizbillah Yazid, Mouli Edward, Mohammad Hardian Basuki, Yunus Abdul Bari, Fedik Abdul Rantam
DOI:10.4103/japtr.JAPTR_52_20  
The advanced, metastasis, and reccurent of osteosarcoma (OS) patients have a poor prognosis postaggresive surgery and chemotherapy. Peripheral blood mononuclear cells (PBMCs) as cell-based immunotherapy may successful in the OS treatment. To investigate the enhancement apoptosis of OS-mesenchymal stem cells (OS-MSCs) co-cultivated with PBMCs sensitized using the secretome and granulocyte macrophage colony-stimulating factor (GMCSF). This true experimental study with posttest only control group design and in vitro study. The sample was cultured OS-MSCs which confirmed by Cluster of Differentiation-133 using immunocytochemistry (ICC) and histopathology analysis. The sample divided into six groups accordingly: OS-MSC, OS-MSC + PMBC, OS-MSC + PMBC + Secretome, OS-MSC + PMBC + GMCSF, OS-MSC + PBMC + Secretome + GMCSF (n = 5/N = 30). The enhancement of OS-MSCs apoptosis was analyzed through Interleukin-2 (IL-2) level through the Enyzme-Linked Immunosorbent Assay examination, expression of Signal Transducers and Activators of Transcription (STAT)-3 and caspase-3 by ICC. One-way analysis of variance test and Tukey Honestly Significant Difference to analyze the difference between the groups (P < 0.05). The highest of IL-2 level was found in the PBMC + Secretome + GMCSF group. The highest expression of caspase-3 was found in OS-MSC + PBMC + Secretome + GMCSF group with significant different between groups (P < 0.05). There was insignificant difference of STAT-3 epxression and IL-2 level between groups (P > 0.05). The co-cultivation of OS-MSCs and PBMSCs activated using secretome and GMCSF has a great ability to enhance OS-MSCs apoptosis.
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Peel-off emulgel mask of Cocos nucifera L. Extract using gelling agent carbomer 940 as antiacne against Propionibacterium acnes ATCC 11827 p. 220
Dewi Melani Hariyadi, Isnaeni Isnaeni, Sisunandar Sudarma, Suciati Suciati, Noorma Rosita
DOI:10.4103/japtr.JAPTR_51_20  
Cocos nucifera Linn., which contain lauric acid has been known had antibacterial activity against Propionibacterium acnes that usually improve severe of pimple. Current study investigated optimum formula of emulgel mask based on the C. nucifera L. Extract from Kopyor coconut. Extract were tested for antibacterial against P. acnes ATCC 11827. In this research, C. nucifera L. extract of 1 and 5% were formulated as an active agent of peel off antiacne emulgel mask-containing carbomer 940 in various concentration (1% and 1.5%). The peel-off emulgel mask of C. nucifera L extract was then evaluated in terms of viscosity, pH, drying time, spreadability, and antibacterial activity. The selected formula was formula containing 5% of extract and 1% of carbomer 940. This formula had pH that suitable with skin pH 4.5–6.5, had good spreadability, and also produced highest antibacterial activity against P. acnes.
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Accelerated wound healing ability of Jatropha sap by iota carrageenan-poly (vinyl alcohol) hydrogel film p. 226
Nurain Thomas, Agus Rusdin, Madania Tulsyahra, Nasrul Wathoni, Asep Kuswandi
DOI:10.4103/japtr.JAPTR_11_20  
Jatropha sap (JTS), an important fluid carried in xylem and phloem tubes of Jatropha multifida L. plant, has good wound healing property. However, physicochemical stability of JTS needs to be improved in order for it to be useful as a topical wound-healing agent. In this study, we developed an iota carrageenan-polyvinyl alcohol (IC-PVA) hydrogel film (HF) as a carrier of JTS and evaluated its wound-healing ability. The characterization of JTS secondary metabolites by ultraviolet-Vis spectrophotometry suggested presence of flavonoid, saponin, and alkaloids in the sap. We successfully extracted IC from Euchima spinosum using alkaline solvent at 80°C–90°C with calcium chloride as the precipitator. The result of computer simulation using Discovery Studio software and Autodock Tools showed the presence of hydrogen bonding interaction of IC-PVA. IC-PVA/JTS HF with excellent physical properties including high swelling ratio (246.32%) and high gel fraction (16.75%). In addition, irritation test in mice confirmed the absence of hypersensitivity reaction, redness, and allergic reactions. Interestingly, IC-PVA/JTS HF significantly accelerated wound healing when compared to the nontreated group/control with 98% wound closure by 10 days. These results suggest that IC-PVA HF has improves wound-healing ability of JTS.
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Induction of cytotoxicity by Bruguiera gymnorrhiza in human breast carcinoma (MCF-7) cell line via activation of the intrinsic pathway p. 233
Gul-e-Saba Chaudhry, Nurul Huda Rahman, Vigneswari Sevakumaran, Aziz Ahmad, Habsah Mohamad, Muhammad Naveed Zafar, Yeong Yik Sung, Tengku Sifzizul Tengku Muhammad
DOI:10.4103/japtr.JAPTR_81_20  
Breast cancer is among the frequently occurring cancer worldwide. The foremost underline aim of this study was to determine the growth inhibitory effect along with mechanistic study of a Bruguiera gymnorrhiza extract on MCF-7. The cytotoxicity activity was determined by using the MTS assay. Butanol extract exhibited the maximum cytotoxicity activity against the MCF-7 cells with IC50of 3.39 μg/mL, followed by diethyl ether and methanol extract (IC50at 16.22 μg/mL and 37.15 μg/mL, respectively) at 72 h. The DeadEndTM Colorimetric Apoptosis Detection System confirmed the induction of apoptosis (via DNA fragmentation) in MCF-7 cells. Both butanol and diethyl ether extracts of B. gymnorrhiza significantly increase the caspase-3 level. However, the diethyl ether extract induced higher caspase-9 levels compared to caspase-8, suggesting that the intrinsic pathway was the major route in the process of apoptosis. Thin-layer chromatography profiling demonstrated the presence of phenolic, terpene, and alkaloid compounds in crude methanol, diethyl ether, and butanol extracts. The phytochemicals present in the extracts of B. gymnorrhiza might have the potential to be a future therapeutic agent against breast cancer.
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