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Backgrounds: Justicia gendarussa Burm.f. Has been known to have anti-HIV activity. This study was conducted to evaluate the effect of incubation time on the antiviral activity of the J. Gendarussa leaves extract on HIV-infected MT-4 cells in vitro. Molecular docking test was also conducted to determine the interaction of alkaloids and flavonoids on the J. Gendarussa leaves against HIV-1 reverse transcriptase receptor. It is expected that this research will provide scientific information on the development of J.
Gendarussa leaves as an anti-HIV drug. Materials and Methods: In the activity test, the effect of incubation time on the antiviral activity of J. Gendarussa leaves on HIV-infected MT-4 cells were evaluated. During the activity test, a parameter of cytolysis effect inhibition on MT-4 cell line was observed after 4 days and 6 days incubation period. The molecular docking test is performed by using Molegro Virtual Docker software to determine the interaction of alkaloid and flavonoid compounds of J.
Gendarussa leaves with HIV-1 reverse transcriptase receptor. Introduction Human Immunodeficiency Virus (HIV) is a retrovirus that infects the human immune system. HIV damages the human immune system by destroying T lymphocyte cell (a type of leukocyte). On the T lymphocyte cell surface, there is cluster of differentiation 4(CD4) receptor. When a number of T-CD4 lymphocyte cells are low then a person more susceptible to be infected (Pattman et al., 2005). The immune system will break down slowly over the increasing number of viruses (viral load) in the body (Naif, 2013). The final stage of HIV infection is AIDS, a condition when CD4 level decrease to very low level (typically.
Plant Materials and Extraction Procedure Leaves of J. Gendarussa were obtained from a cultivated crop in Pacet, Mojokerto, East Java province, Indonesia. This plant was identified by the Department of Pharmacognosy and Phytochemistry, Faculty of Pharmacy, Airlangga University under the voucher number 18/H3.1.5/DT/2012. Simplicia powder of J. Gendarussa leaves was divided into two groups: acidified leaves powder to release alkaloids and non-acidified leaves powder. Both the powder was extracted using 70% ethanol for 3x24 hours in a macerator device, and then the resulting filtrate were concentrated using a rotary evaporator.
The extract is dried at 50°C a temperature of to obtain a 70% ethanol extract (17.4% w/w) and fractionated-70% ethanol extract (6.4% w/w) of J. Gendarussa leaves. Water extract of leaves of J. Gendarussa is made by blending fresh J. Gendarussa leaves in cold water.
Then the resulted filtrate was collected and was dried using the freeze-dry method to obtain water extracts (1.8% w/w) of J. Gendarussa leaves. Mojo master winamp plugin. Alkaloid and Flavonoid Screening of J. Gendarussa Leaves Extract Alkaloid screening of 70% ethanol extract, fractionated-70% ethanol extract and water extract of J. Gendarussa leaves be performed using a TLC GF254 plate with mobile phase chloroform: methanol (9:1). Here, Dragendorf reagent is used to reveal stains. Results test is said to be alkaloid positive if there are orange stains.
Flavonoid screening of 70% ethanol extract, fractionated-70% ethanol extract and water extract of J. Gendarussa leaves be performed by using the immobile phase of TLC GF254 with a mobile phase of butanol: glacial acetic acid: water (4:1:5). Here, borate citric is used to reveal stains. Results test is said to be flavonoid acid if there are yellow-green fluorescence stains using UV 366 nm (Indonesia, 2008). Reagents and Chemicals Zidovudine-Lamivudine (ZDV/3TC) (Duviral ®), 2 µm nitrocellulose membrane filter (Whatman), sterile water for injection, and aquadest were obtained from CRS-EIRD, Institute of Tropical Disease, Surabaya, Indonesia.
RPMI-1640 medium and Fetal Bovine Serum (FBS) were purchased from Gibco. Natrium bicarbonate and 70% ethanol pharmaceutical grade were purchased from Merck. WST-1 was purchased from Roche. Dimethyl sulfoxide (DMSO) was purchased from Sigma.
Cells and Viruses The MT-4 cells (human T- cell leukemia line) were obtained from the Institute of Tropical Disease (ITD) laboratory, Airlangga University, Surabaya, Indonesia. MT-4 cells were cultured in RPMI-1640 media and were equipped with 10% FBS. The MT-4 cells were maintained in T 25 CCF at 37 °C temperature in a 5% CO 2 incubator. HIV isolates from a seropositive HIV donor that labeled IDU-18 obtained from ITD laboratory, Airlangga University, Surabaya, Indonesia. HIV cultured on the MT-4 cell in RPMI-1640 medium completed with FBS 10%. MT-4/HIV cell kept in CCF T 25 at 37 °C in CO 2 5% incubator.