DOI: 10.14457/tu.the.2016.1481
title: Anti-microbial, anti-inflammatory and anti-estrogenic activities of “Sa-Tri-Lhang-Klod” remedy for postpartum care

description: Sa-Tri-Lhang-Klod (STK) is Thai traditional medicine preparation, is used for care during postpartum phase. It has been used for postpartum care on excrete lochia, blood tonic, uterine involution and improving blood circulation. Sa-Tri-Lhang-Klod remedy comprises 17 plant ingredients consists of roots of Maclura cochinchinensis (Lour.) Corner, stems of Artocarpus heterophyllus Lam., rhizomes of Curcuma comosa Roxb., stems of Caesalpinia sappan Linn., vines of Piper ribesioides Wall., roots of Plumbago indica Linn., flowers of Piper longum Linn., roots of Angelica sinensis (Oliv.) Diels., stems of Salacia chinensis Linn., fruits of Piper nigrum Linn., roots of Piper samentosum Linn., flowers of Carthamus tinctorius Linn., flowers of Jasminum sambac Ait., flowers of Mimusops elengi Linn., flowers of Mesua ferrea Linn., flowers of Mammea siamensis Kosterm., and flowers of Nelumbo nucifera Gaertn. The taste of these preparation is hot and spicy. Sa-Tri-Lhang-Klod remedy and some plant ingredients in this remedy have been investigated for antioxidant, cytotoxicity and anti-inflammatory. However, for Sa-Tri-Lhang-Klod remedy, there is no report to study on antimicrobial, anti-inflammatory (COX-2) and estrogenic effect. Therefore, the objective of this study was to investigate the antimicrobial, anti-inflammatory (COX-2) and estrogenic effect of STK extracts and plant ingredients.STK and its plant ingredients were extracted by maceration in 95% ethanol, 50% ethanol and decoction in water. All extracts of 20 samples were tested for, the first, antimicrobial activity against Staphylococcus aureus (ATTC 25923), Escherichia coli (ATTC 25922), and Candida albicans (ATTC 90028) by disc diffusion method. Minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) of all extracts were measured by broth dilution method. The results found that the inhibition zone of STK95 for inhibit S. aureus and C. albicans values 9.33±0.58 and 7.33±0.58 mm, respectively. STK50 and STW can inhibit S. aureus at 10.00±0.00 mm and 7.00±0.00 mm. And the ethanolic extract of C. sappan exhibited antimicrobial activity against S. aureus and E. coli with the highest inhibition zone of 22.67±2.32 mm and 10.33±0.58 mm, respectively. 95% ethanolic extract of P. indica exhibited the highest antimicrobial activity against C. albican with the highest inhibition zone of 13.00±0.00 mm. 95% ethanolic extract of M. siamensis showed bacteriacidal activity against S. aureus with MIC and MBC of 0.019 mg/ml. Moreover, the MIC and MBC values for 95% ethanolic extract of C. sappan against E. coli and C. albican were observed to be same values (1.25 mg/ml and 0.625 mg/ml, respectively).For the anti-inflammatory, the 95% ethanolic extract (STK95) showed anti-inflammatory activity of inhibitory effect on cyclooxygenase-2 enzyme (Cox-2) in RAW 264.7 cells with IC50 of 3.15±0.30 µg/ml. However, it was less than prednisolone (positive control) with IC50 value of 1.34±0.03 µg/ml. While, the 95% exthanolic extract of M. siamensis showed the higher anti-inflammatory activity than prednisolone with IC50 value of 0.08±0.005 µg/ml.For the estrogenic and anti-estrogenic activities against E2-enhance T47D and MCF-7 cell proliferation found STK95, STK50 and STW showed anti-estrogenic activity. STK50 showed strong inhibition against T47D and MCF-7 cells at low concentration with iEqE1 values &lt; 0.01 µM which it against more antiestrogenic effect than tamoxifen (iEqE1=1.79 and 84.56 µM).The 95% ethanolic extract of M. siamensis was chosen to separate by Vacuum Liquid Chromatography (VLC) because it showed good activity in many bioassays. It was separated by vacuum liquid chromatography to be 5 fractions; Hexane (MS-1), Hexane:Chloroform (MS-2), Chloroform (MS-3), Chloroform:Methanol (MS-4) and Methanol (MS-5). MS-4 fraction which showed the highest activities was isolated active compound by using column chromatography. The component of coumarin was isolated. It showed the anti-inflammatory in RAW 264.7 cells (COX-2) values 47.27±2.00 µg/ml and had no toxicity.In conclusion, STK95, STK50 and STKW showed the moderate of antimicrobial. M. siamensis showed very high biological in antimicrobial and anti-inflammatory. And the all extract of remedy showed the good of anti-estrogenic activity. However, the results of the present study can partially support the use of STK, a remedy in Thai traditional medicine for protection of postpartum infection, anti-inflammation after delivery and for increasing lactation. Moreover, these results related to Thai traditional medicine and to support the using these remedy in the lists of The National List of Essential Herbal Medicine (NLEM) of Thailand.