Effects of Labisia pumila VAR. alata and VAR. pumila leaf aqueous extract cream on wound healing using rat models

Abstract

Labisia pumila (L. pumila) or locally known as Kacip Fatimah is a traditional herbal medicine commonly used in women related treatments. Although there are many researches conducted to relate this plant species with many health and medicinal benefits, L. pumila’s potential as a wound healing agent has not been explored in the aspect of non-clinical studies. This study was done to establish the wound healing properties of two varieties of L. pumila var. alata and var. pumila in rats. LC-MS was used for metabolite analysis on samples obtained from the cream preparation phase of L. pumila aqueous extract. Skin tissues of 48 male and 48 female Sprague Dawley rats were wounded with 6 mm biopsy punches and wound area were measured at day two, five and eight while RNA expression of selected genes from the inflammation (TGF-1, IL-6), proliferation (EGF, CBL) and remodelling (COL3A1) phases of the wound healing mechanism were measured using qPCR. Proteins of rat wound tissue were identified using LC-MS protein analysis. Three metabolites which were retained throughout the cream preparation were identified as tyrosol, protocatechuic acid and catechin. These metabolites have already been reported to have properties beneficial to the wound healing mechanism. Wounds of female rats treated with var. alata healed in 12 days and var. pumila in 13 days where var. alata had significant healing on day 2 (79.58% ± 0.55) and var. pumila was significant on day 5 (75.97% ± 0.90) as well as day 8 (95.04% ± 0.26). For male rats, wounds treated with var. pumila healed in 12 days and var. alata in 13 days while control took 17 days. Gene expression studies in female var. alata samples show gene expression was upregulated during the cell proliferation (46136 mRNA copy number ± 2942) and tissue remodelling phases (2.44x108 mRNA copy number ± 23210943) while for male var. alata group genes were upregulate in inflammation (1722784 mRNA copy number ± 88233) and remodelling phase (3.91x108 mRNA copy number ± 14296340). For var. pumila treatment groups, gene expression was downregulated at all phases of wound healing. Protein analysis through LC-MS was able to identify 133 proteins from the skin of female rats treated with var. alata. Proteins which contributed to the certain wound healing phases were found in abundance on days specific to the phase. Protein elastase-2, neutrophil was found in abundance on day 2 as it contributes to the inflammation phase. On day 5, protein disulphide-isomerase A6 that is important in cell proliferation is found in abundance while collagen alpha-1(1) chain, a protein vital in the remodelling phase is found in abundance on day 8. There results from this study show that var. alata and var. pumila contribute to wound healing in both male and female rats but might take part if upregulation and down regulation of different genes for the different genders. The information and data obtained from this study will be able to contribute to the pharmaceutical industry and furthermore will increase the potential of local herbs in the wound care and management in Malaysia.