| Summary: | Purpose: This study aims to examine the ability of ethanol extract and essential oil of
red betel vine leaves as antimycobacterium against Mycobacterium tuberculosis both
in vitro and in vivo. In addition, it also aims to determine its safety by conducting
acute toxicity tests.
Method: The design of this research is an experimental laboratory for in vitro tests
and true experimental for in vivo tests and acute toxicity tests. The in vitro study uses
a serial dilution method, by finding the minimum bactericidal concentration (MBC).
The concentrations of ethanol extract tested are 500 mg/mL, 400 mg/mL, 250 mg/mL,
200 mg/mL, 125 mg/mL, 100 mg/mL, and 62.5 mg/mL. Meanwhile, the
concentrations of essential oils tested are 360 mg/mL, 180 mg/mL, 90 mg/mL, 45
mg/mL, 22.5 mg/mL, 11.25 mg/mL, 5.63 mg/mL, and 2.82 mg/mL. The influence of
ethanol extract and essential oil toward cell wall of M. tuberculosis is observed with
Transmission Electron Microscope (TEM). The analysis was done descriptively. In
the in vivo test, the infection was done with intratracheal method. The mice were
classified into 8 groups with each group consisting of 6 mice (3 female and 3 male).
The infection and therapy were done for 15 days for each process. Group I was given
1.64 mg ethanol extract (54.67 mg/kg body weight), group II was given a dose of 8.2
mg (273.35 mg/kg), group III was given a dose of 41 mg (1366.67 mg/kg). Group IV
was given a dose of 0.06 mg of essential oil (2 mg/kg), group V was given a dose of
0.3 mg (10 mg/kg), and group VI was given a dose of 1.5 mg (50 mg/kgBB). Group
VII as the control group received aquades. Group I-VII received twice a day therapy
in the morning and afternoon. Group VIII received standard drug with one time
consumption a day in the morning time. In addition, the uninfected group became the
health control. The treatment is done for 15 days. After that, mice were terminated
and their lungs were taken. Afterward, slides were made by Ziehl Neelsen staining
(ZN) and Haematoxylin & eosin (HE). The analysis of ZN and slides were done with
Kruskal Wallis statistic test and it was continued with U Mann Whitney test.
However, HE slides should be previously assessed with a Dormans score criteria. The
effective dose was calculated based on probit. Acute toxicity test is calculated based
on the formula of Karber.
Result: In the in vitro examination, the MBC value of red betel vine leaves ethanol
extract is obtained in the concentration of 100 mg/mL. Meanwhile, the value of MBC
for essential oil is obtained in the concentration of 5.63 mg/mL. The observation
using TEM, damage in the cell wall was found both in the exposure of ethanol extract
and essential oil of red betel vine leaves. In the in vivo test, the ethanol extract of red
betel vine leaves can decrease the bacteria M. Mycobacterium up to 89.53% with an
optimal dose of 8.2 mg (273.35 mg/kg), while the essential oils can reduce the
bacteria of M. Mycobacterium up to 89.56%, with the optimal dose of 0.3 mg (10
mg/kg bodyweight). The median effective dose for the ethanol extract 125.13 mg/kg.
However, the median effective dose for essential oils could not be determined in this
study. HE slides results obtained best dose to 8.2 mg ethanol extract (273.35 mg/kg)
and for the essential oil of 0.3 mg (10 mg/kg) but it has not been statistically proven.much as 18,000 mg/kg body weight for ethanol extract and for 11,700 mg/kg for
essential oil.
Conclusion: The ethanol extract and essential oil of red betel vine leaves can function
as antimycobacterium both in vitro and in vivo. The ethanol extract and essential oil
of red betel leaves are safe for consumption.
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