Fibroblast and keratinocyte gene expression following exposure to the extracts of holy basil plant (Ocimum tenuiflorum), malabar nut plant (Justicia adhatoda), and emblic myrobalan plant (Phyllanthus emblica)

This data article provides gene expression profiles, determined by using real-time PCR, of fibroblasts and keratinocytes treated with 0.01% and 0.001% extracts of holy basil plant (Ocimum tenuiflorum), sri lankan local name “maduruthala”, 0.1% and 0.01% extracts of malabar nut plant (Justicia adhatoda), sri lankan local name “adayhoda” and 0.003% and 0.001% extracts of emblic myrobalan plant (Phyllanthus emblica), sri lankan local name “nelli”, harvested in Sri Lanka. For fibroblasts, the dataset includes expression profiles for genes encoding hyaluronan synthase 1 (HAS1), hyaluronan synthase 2 (HAS2), hyaluronidase-1 (HYAL1), hyaluronidase-2 (HYAL2), versican, aggrecan, CD44, collagen, type I, alpha 1 (COL1A1), collagen, type III, alpha 1 (COL3A1), collagen, type VII, alpha 1 (COL7A1), matrix metalloproteinase 1 (MMP1), acid ceramidase, basic fibroblast growth factor (bFGF), fibroblast growth factor-7 (FGF7), vascular endothelial growth factor (VEGF), interleukin-1 alpha (IL-1α), cyclooxygenase-2 (cox2), transforming growth factor beta (TGF-β), and aquaporin 3 (AQP3). For keratinocytes, the expression profiles are for genes encoding HAS1, HAS2, HYAL1, HYAL2, versican, CD44, IL-1α, cox2, TGF-β, AQP3, Laminin5, collagen, type XVII, alpha 1 (COL17A1), integrin alpha-6 (ITGA6), ceramide synthase 3 (CERS3), elongation of very long chain fatty acids protein 1 (ELOVL1), elongation of very long chain fatty acids protein 4 (ELOVL4), filaggrin (FLG), transglutaminase 1 (TGM1), and keratin 1 (KRT1). The expression profiles are provided as bar graphs.

The data in this article provides useful knowledge for the cosmeceutical application of holy basil extract, malabar nut extract and emblic myrobalan, traditional ayurvedic plants in Sri lanka.

Materials
Holy basil plants (Ocimum tenuiflorum) were harvested from a medicinal garden at the Institute of Traditional Plants in Sri Lanka (Negombo, Sri Lanka). The plant shoot metabolites were extracted by using 70% ethyl alcohol solution. Malabar nut plants (Justicia adhatoda) were harvested from a medicinal garden at the Institute of Traditional Plants in Sri Lanka (Negombo, Sri Lanka). The plant leave metabolites were extracted by using 70% ethyl alcohol solution. Emblic myrobalan plants (Phyllanthus emblica) were harvested from a medicinal garden at the Institute of Traditional Plants in Sri Lanka (Negombo, Sri Lanka). The plant leave metabolites were extracted by using 50% ethyl alcohol solution.

Fibroblast cell culture
Normal human skin fibroblasts, RIKEN original (NB1RGB), were provided by the RIKEN BRC through the National Bio-Resource Project of the MEXT, Japan. The cells were cultured in Minimum Essential Media-alpha (MEMα; Life Technologies Corp.) supplemented with 10% fetal bovine serum   (FBS; Biowest) and 0.2% NaHCO3. The cells were grown at 37°C in a humidified incubator containing 5% CO 2, according to the manufacturer's instructions. For all of the experiments, human fibroblasts were seeded into a 60 mm dish (5 × 10 4 cells/dish) and incubated for 8 h with culture media containing 10% FBS. The cells were subsequently subjected to serum starvation for 16 h with serum-free MEMα.

Keratinocyte cell culture
Normal human epidermal keratinocytes (HEKn; GIBCO) were isolated from neonatal foreskin. The cells were cultured in Medium 154 (Invitrogen) supplemented with human keratinocyte growth factor (HKGS; Invitrogen), according to the manufacturer's instructions. The cells were grown at 37°C in a humidified incubator containing 5% CO 2 . For all of the experiments, human keratinocytes were seeded into a collagen-coated 60 mm dish (5 × 10 4 cells/dish), and incubated for 8 h with culture media containing HKGS. The cells were next subjected to HKGS starvation for 16 h with Medium 154.

Exposure of the cells to the plant extract, RNA isolation and quantitative real-time PCR
The cells were seeded into a 60 mm dish (5 × 10 4 cells/dish). The cells were exposed to 0.01% or 0.001% of the plant extract, for 24 h at 37°C. The cells were collected at 2, 4, 8, and 24 h after initiation of the exposure. Total RNA was extracted from the cells by using the TRI reagent (Merck). This RNA extract was used as a template for subsequent cDNA synthesis with oligo dT primers (Table 1), using the Primescript RT reagent Kit (Takara bio inc.). The mRNA levels were quantified using a LightCycler 96 system (Roche) and SYBR Premix Ex Taq II (Takara Bio Inc.). The data were analyzed using the delta  cycle threshold method, and calculated based on the Cq values, and the expression of each gene was normalized to GAPDH. All values are reported as means 7 standard error, as previously described [18].

Statistical analysis
All the values have been reported in terms of mean 7 SE values. The data were analyzed using the Student's t-test. A P value less than 0.05 was considered to be statistically significant.