Original Articles: 2013 Vol: 5 Issue: 5
Fluorescence of Senna simea Lam. leaf extracts: A possible interference in a fluorescence-based assay
Abstract
Major depressive disorder (MDD) is a debilitating disease worldwide and occurs with a high prevalence in elderly
individuals. In attempt to search for herbal medicine with antidepressant effect, each in vitro model must be
properly validated. The objective of this study was to determine the effect of Senna siamea Lam. (Cassod tree) leaf
extracts prepared using ethanol and water on activity of DAT, NET, SERT and total MATs in in vitro model, human
neuroblastoma LAN-5 cell line using a commercial neurotransmitter transporter reuptake assay kit. After measuring
the intracellular fluorescence, we calculated RFU area below curve reflecting the activity of dopamine transporter
(DAT), norepinephrine transporter (NET), serotonin transporter (SERT) and all monoamine neurotransmitter
transporters (MATs) in LAN-5 cells after treatment for 30 min with 25 and 100 μg/mL of Senna siamea Lam. leaf
extracts in the absence or presence of combining antidepressants. We observed the fluorescence intensity below zero
or minus values, thus reflecting assay interference. The inference was stronger as reflected by lesser intracellular
fluorescence intensity when increasing concentrations of Senna siamea Lam. extracts. Ethanol extract resulted in a
stronger interference as compared to that of water extract. To confirm that Senna siamea Lam. leaf extracts
containing certain fluorescent compounds with excitation 440 nm and emission 520 nm, we measured the
fluorescence of Senna siamea extracts compared with reagent blank and Mentha cordifolia Opiz ex. Frezen (Kitchen
mint) leaf and Centella asiatica L (Urm.) (Gotu kola) whole plant extracts as control herbal extracts using
spectrofluorometer with the excitation wavelength of 440 nm and emission wavelengths of 480-600 nm for scanning,
and we found the concentration-dependent and solvent type-dependent manner in agreement with the observation in
neurotransmitter transporter reuptake assay. Therefore, Senna siamea leaf extracts possessed fluorescence
characteristics possibly due to certain fluorescent compounds and could be interfered in a fluorescence-based
assay. It is highly recommended to know the chemical nature of each plant extract before selecting any biological
assay.