Cytotoxic Activity of Ethanolic Extract Aquilaria malaccensis Leaves Against MCF-7 Cells

@#Introduction: Aquilaria malaccensis, also known as “Pokok Karas” in Malaysia, is widely used in Southeast Asian countries for the treatment of joint pain, diarrhoea and inflammatory diseases, and has shown beneficial effects as an anticancer agent. The aim of this study was to investigate the effect of ethanol leaf extracts of A. malaccensis on MCF-7 cells. Methods: MTT-based cytotoxic and antiproliferative assay was used to determine the outcome of ethanolic extract toward MCF-7 cells. The mode of cell death was determined by the AO/PI double staining assay and the depolarisation of the mitochondria membrane potential. Results: IC50 value of the extract against MCF-7 cells treated for 72 hours was 4.1 ± 2.08 µg/mL, while the IC50 value for doxorubicin was 2.92 ± 0.12 µg/mL. The extract showed a lower cytotoxic effect against the NIH/3T3 cells and inhibited the growth of MCF-7 cells in a dose dependent manner. AO/PI double stain showed that the ethanolic extract of A. malaccensis leaves induced MCF-7 cells into apoptotic cell death. The present study showed that the ethanolic extract of A. malaccensis induced apoptosis through mitochondrial pathway as indicated by its ability to take up JC-1. Conclusion: The study found that ethanolic extract obtained from A. malaccensis leaves is cytotoxic on MCF-7 cells, resulting to apoptotic cell death of the cells.

Isolation and morphological identification of endophytic fungi from leaves and stems of Aquilaria malaccensis Lamk. originating from Java Island, Indonesia

Aims@#Endophytic fungi are the remarkable category of host-associated fungal community that invades the intercellular regions of host tissues, benefiting their host while obtaining an advantage. Fungal endophytes have lately attracted prominence as a source of active secondary metabolites. This investigation aimed to identify fungal endophytes that reside inside the leaves and stems of Aquilaria malaccensis.@*Methodology and results@#Healthy A. malaccensis stems and leaves samples were collected. Clean leaves and stems were cut to a size of 1 cm, followed by sterilization using 75% ethanol for 1 min, 3% sodium hypochlorite solution for 1 min, and finally, rinsing with sterile water 3 times for 1 min and drying with sterile paper. The sterile samples were put onto Potato Dextrose Agar (PDA) media containing chloramphenicol for 7-14 days until the mycelium grew for morphological identification under a light microscope. Five endophytic fungi were recovered from leaves, while nine endophytic fungi were obtained from stems. Using morphological approaches, nine of the endophytes had observed to produce conidia fungi, whereas the others did not. Neopestalotiopsis sp., Aspergillus sp., Arthrinium sp., Curvularia sp., Podospora sp., Mucor sp. and Verticillium sp. were identified as nine of the fourteen endophytes.@*Conclusion, significance and impact of study@#The number of endophytic fungi discovered in different organs varies. Not all endophytic fungi that grow can create sexual phages. Six genera of endophytic fungi were identified.

Combination of polymyxin B and Aquilaria malaccensis extract enhanced the killing and inhibited the growth of Acinetobacter baumannii and Klebsiella pneumoniae

Aims@#Polymyxins are an important last-line treatment for infections caused by multidrug-resistant Gram-negative bacteria. Nonetheless, the emergence of polymyxin-resistance and the limiting of polymyxin monotherapy urgently demands its optimisation. Aquilaria malaccensis (Agarwood) has been widely used as traditional medicine. Many parts of the plant including leaves exhibit a considerable in vitro antibacterial activity against microbial pathogens. Exploiting A. malaccensis in combination with polymyxins provides a novel strategy in fighting antimicrobial resistance. The objective of this study was to evaluate the combination effects of A. malaccensis extract with polymyxins against Acinetobacter baumannii and Klebsiella pneumoniae.@*Methodology and results@#In vitro time-kill studies and GC-MS analysis were performed to evaluate the bacterial killing of polymyxin B and extract combination and analyse chemical compounds of the extract, respectively. The combination of polymyxin B (1 mg/L) and A. malaccensis extract (32 mg/mL and 64 mg/mL) treatments exhibited enhanced bacterial killing compared to polymyxin B alone at 4 h and 24 h. Combination treatments also inhibited the bacterial growth of both A. baumannii and K. pneumoniae observed throughout the 24 h. More than sixty compounds including phytol, 9,12-octadecadienal, fatty acid, alkanes and terpenoids were putatively identified as the compounds that likely contributed to the antibacterial activity.@*Conclusion, significance and impact of study@#This study was the first to report the potential application of A. malaccensis extract in combination with polymyxin B in treatment against A. baumannii and K. pneumoniae and can be further investigated and optimized for the treatment of bacterial infectious diseases.

Evaluation of Acute and Sub-acute Oral Toxicity Effect of Aquilaria malaccensis Leaves Aqueous Extract in Male ICR Mice

The study was conducted to investigate the acute and sub-acute toxicity effect of Aquilaria malaccensis leaves aqueous extract (AEAM) towards male ICR mice in terms of body weight, relative organ weight, mortality rate and sperm parameters. In acute toxicity study, a single dose at of 2000 mg/kg was performed. In sub-acute toxicity study, the mice were received normal saline (control group), 50, 100, 150, 200, 500, or 1000 mg/kg of AEAM orally for 21 days of treatment. In sub-acute toxicity study, the number of abnormal sperm were significantly decreased in AEAM 100, 150, 200, 500, and 1000 when compared to the control group. While, the motility of sperm were found to be significantly increased in AEAM 100, 150, 200, and 1000 as compared to the control group. No mortality was recorded in the control group and treated groups in both toxicity studies except for one mouse from AEAM 1000 group. However, the mild sedative effect in terms of the tendency to sleep was clearly noticeable in both toxicity studies. Results indicated that the AEAM can be one of the useful alternative medicine to enhance fertility rate by increasing healthy sperm production.

Ameliorative Effects of Aquilaria malaccensis Leaves Aqueous Extract on Reproductive Toxicity Induced by Cyclophosphamide in Male Rats

@#Background: Cyclophosphamide (CP) is a widely used anti-neoplastic and immunosuppressive agent that is associated with adverse side effects including reproductive toxicity. Aquilaria malaccensis (AM) is a traditional medicinal plant which was reported to exhibit high anti-oxidant and free radical scavenging properties. The present study was aimed to evaluate the protective effects of AM leaves extract on sperm quality following toxic exposure to CP. Methods: Forty-eight male Sprague Dawley rats were allocated into eight groups of six rats (n = 6): control, CP only (200 mg kg−1), AM only (100 mg kg−1, 300 mg kg−1 and 500 mg kg−1) and CP + AM (100 mg kg−1, 300 mg kg−1 and 500 mg kg−1). Animals were sacrificed after 63 days of treatment and the sperm from the caudal epididymis was taken for sperm analysis. Results: The body and the reproductive organs weight, sperm count and motility did not differ between CP and other groups (P > 0.05). A significant increase (P < 0.05) in percentage of the dead and abnormal sperm were seen in the CP alone treated group compared to the control group. Co-administration of AM to the CP exposed rats significantly reduced the (P < 0.05) percentage of abnormal sperm as compared to the CP only group. Conclusion: Overall, the present results represent the potential of AM to protect against CP induced reproductive toxicity.

The Effects of Aquilaria malaccensis Leaves Aqueous Extract on Sperm of Sprague Dawley Rats towards Early Embryogenesis

@#Introduction: Oxidative stress induced by excessive and unopposed levels of reactive oxygen species in male reproductive system results in impaired sperm quality, fertilization capacity and poor embryo development. Our goal is to assess the potential effects of Aquilaria malaccensis (AM) leaves, a plant with strong antioxidant property on early embryo development in vitro and embryo quality following fertilization with cyclophosphamide (CP) exposed rat sperm. Materials and Methods: Twenty four male Sprague Dawley rats were allocated into eight groups of three rats (n = 3): control, CP only (200 mg/kg), AM only (100 mg/kg, 300 mg/kg and 500 mg/kg) and CP + AM (100 mg/kg, 300 mg/kg and 500 mg/kg). Animals were sacrificed after 63 days of treatment and sperm from caudal epididymis were taken for in vitro fertilization (IVF) with oocytes from untreated female. Fertilization, embryo division and embryo morphology were examined at 8 and 48 hours post insemination and compared between groups. Statistical evaluations were performed using Chi-Square test and Fisher’s exact test and p-value<0.05 was considered significant. Results: Administration of AM leave extract at 100 mg/kg/day to normal rats and CP-exposed rats has significantly increased (p<0.05) the fertilization rate, early cleavage rate and embryo quality when compared to CP only treated group. However, other groups showed no significant differences. Conclusion: Overall, the present results indicate the potential of AM leave extract supplementation to improve the fertility and early embryo development in male rat exposed to CP by inhibiting the oxidative processes and scavenging free radicals.