Exploring in vitro anti-proliferative and anti-inflammatory activities of Prasachandaeng remedy, and its bioactive compounds.

BACKGROUND: Prasachandaeng (PSD) remedy has been empirically used in Thai traditional medicine to treat fever in bile duct and liver and cancer patients through Thai folk doctors. However, there have been no scientific reports on the bioactive compounds and bioactivities related to inflammation-associated carcinogenesis or cytotoxicity against cancer cell lines. In this study, we investigated the chemical content of the remedy, and evaluated its cytotoxic activity against two cancer cell lines in comparison with a non-cancerous cell line and determined tumor necrosis factor-alpha (TNF-α) production in a murine macrophage cell line (RAW 264.7) to evaluate anti-inflammatory activity. A novel HPLC method was used for quality control of its chemical content. METHODS: Pure compounds from the EtOH extract of D. cochinchinensis were isolated using bioassay-guided fractionation and chemical content of the PSD remedy was determined using HPLC. The cytotoxic activity against the hepatocarcinoma cell line (HepG2) and cholangiocarcinoma cell line (KKU-M156), in comparison with non-cancerous cell line (HaCaT), were investigated using antiproliferative assay (SRB). The anti-inflammatory activity measured by TNF-α production in RAW 264.7 was determined using ELISA. RESULTS: All crude extracts and isolated compounds exhibited significant differences from vincristine sulfate (****p < 0.0001) in their cytotoxic activity against HepG2, KKU-M156, and HaCaT. The PSD remedy exhibited cytotoxic activity against HepG2 and KKU-M156 with IC50 values of 10.45 ± 1.98 (SI = 5.3) and 4.53 ± 0.74 (SI = 12.2) µg/mL, respectively. Some constituents from C. sappan, D. cochinchinensis, M. siamensis, and M. fragrans also exhibited cytotoxic activity against HepG2 and KKU-M156, with IC50 values less than 10 µg/mL. The isolated compounds, i.e., Loureirin B (1), 4-Hydroxy-2,4'-dimethoxydihydrochalcone (2), and Eucomol (3) exhibited moderate cytotoxicity against two cancer cell lines. None of the crude extracts and isolated compounds showed cytotoxicity against HaCaT. D. cochinchinensis and PSD remedy exhibited higher anti-inflammatory activity measured as TNF-α production than acetaminophen. CONCLUSION: The findings provide evidence of bioactivity for EtOH extracts of PSD remedy and the isolated compounds of D. Cochinchinensis. The results consistent the use clinical activity and use of PSD remedy as a antipyretic treatment for liver and bile duct cancer patients by Thai traditional practitioners.

Anti-Inflammatory Investigations of Extracts of Zanthoxylum rhetsa.

Zanthoxylum rhetsa has been consumed in the diet in northern Thailand and also used as a medicament in ancient scripture for arthropathies. Thus, this study aimed to evaluate the activity of various extracts from differential parts of Z. rhetsa via inhibition of inflammatory mediators (NO, TNF-α, and PGE2) in RAW264.7 macrophages. The chemical composition in active extracts was also analyzed by GC/MS. The parts of this plant studied were whole fruits (F), pericarp (P), and seed (O). The methods of extraction included maceration in hexane, 95% ethanol and 50% ethanol, boiling in water, and water distillation. The results demonstrated that the hexane and 95% ethanolic extract from pericarp (PH and P95) and seed essential oil (SO) were the most active extracts. PH and P95 gave the highest inhibition of NO production with IC50 as 11.99 ± 1.66 µg/ml and 15.33 ± 1.05 µg/ml, respectively, and they also showed the highest anti-inflammatory effect on TNF-α with IC50 as 36.08 ± 0.55 µg/ml and 34.90 ± 2.58 µg/ml, respectively. PH and P95 also showed the highest inhibitory effect on PGE2 but less than SO with IC50 as 13.72 ± 0.81 µg/ml, 12.26 ± 0.71 µg/ml, and 8.61 ± 2.23 µg/ml, respectively. 2,3-Pinanediol was the major anti-inflammatory compound analyzed in PH (11.28%) and P95 (19.82%) while terpinen-4-ol constituted a major anti-inflammatory compound in SO at 35.13%. These findings are the first supportive data for ethnomedical use for analgesic and anti-inflammatory activity in acute (SO) and chronic (PH and P95) inflammation.

Ethnopharmacological analysis from Thai traditional medicine called prasachandaeng remedy as a potential antipyretic drug.

ETHNOPHARMACOLOGICAL RELEVANCE: Prasachandaeng (PSD) remedy is a famous antipyretic drug for adults and children in Thai traditional medicine used and is described in Thailand's National List of Essential Medicine. Relationship between the taste of this herbal medicine, ethnopharmacological used and its pharmacological properties was reviewed. AIMS OF STUDY: Since there has been no scientific report on the antipyretic activity of PSD, aim of this study was to investigate the efficacy related antipyretic drug of the remedy and its 12 herbal ingredients. It involved quality evaluation of raw materials, extraction of PSD and its ingredients, in vitro evaluation of their inhibitory activities on fever mediators, i.e. NO and PGE2 production in murine macrophage (RAW 264.7) cell line stimulated by lipopolysaccharide, and its stability study of the 95% ethanolic extract of PSD remedy. MATERIALS AND METHODS: PSD remedy was extracted by maceration with 50% and 95% ethanol (PSD50 and PSD95), by decoction with distilled water (PSDW), and hydrolysis of PSDW with 0.1 N HCl (PSDH). The 12 plant ingredients were extracted with 95% ethanol. Quality evaluation of PSD ingredients was performed according to the standard procedures for the quality control of herbal materials. The inhibitory activity on nitric oxide production was determined by the Griess reaction and the inhibition of prostaglandin E2 production was determined using the ELISA test kit. RESULTS: PSD ingredients passed the quality standard stipulated for herbal materials. PSD95 exhibited the highest inhibitory activities on the production of NO and PGE2 with the IC50 values of 42.40 ± 0.72 and 4.65 ± 0.76 µg/mL, respectively. A standard drug acetaminophen (ACP) exhibited inhibition of NO and PGE2 production with the IC50 values of 99.50 ± 0.43 and 6.110 ± 0.661 µg/mL, respectively. The stability study was suggested two years shelf-life of PSD95. This is the first report on the activity related antipyretic activity of PSD remedy and its ingredients against two fever mediators, NO and PGE2. CONCLUSION: The results suggested that the 95% ethanolic extracts of PSD remedy and some of its ingredients, were better than ACP in reducing fever. PSD should be further studied using in vivo models and clinical trials to support its use as an antipyretic drug in Thai traditional medicine.

The anti-allergic and anti-inflammatory effects of Benjakul extract (a Thai traditional medicine), its constituent plants and its some pure constituents using in vitro experiments.

Benjakul (BJK), a Thai traditional medicine preparation, has long been used for balanced health, controlled abnormal of element in the body, carminative, and relief of flatulence. It is composed of five plants: Piper interruptum Opiz., Piper longum L., Piper sarmentosum Roxb., Plumbago indica L., and Zingiber officinale Roscoe. The ethanolic extracts of BJK, its five individual plants, and pure constituents of BJK were investigated for their anti-allergic activity using immunoglobulin E (IgE)-sensitized ß-hexosaminidase in the rat basophilic leukemia-2H3 (RBL-2H3) cells and anti-inflammatory activity using lipopolysaccharide (LPS)-induced nitric oxide (NO) and tumor necrosis factor-alpha (TNF-α) in the murine macrophage (RAW 264.7) cells. The ethanolic extracts of BJK showed anti-allergic activity (IC50=12.69µg/ml) and exhibited potent NO inhibitory effect (IC50=16.60µg/ml), but inactive on TNF-α release. Moreover, 6-shogaol and plumbagin, two pure compounds from BJK, showed higher anti-allergic activity than the ethanolic BJK extract with IC50 values of 0.28 and 4.03µg/ml, respectively. These compounds were significantly higher than chlorpheniramine (CPM), standard drug, with IC50 value of 17.98µg/ml. Determination of the anti-inflammatory activity by measuring the inhibition of NO production presented that plumbagin and 6-shogaol exhibited higher than crude BJK extract with IC50 values of 0.002 and 0.92µg/ml, respectively. In particular, plumbagin also showed higher anti-inflammatory than prednisolone, positive control, with IC50 value of 0.59µg/ml. 6-Shogaol also showed inhibitory effect on TNF-α release (IC50=9.16µg/ml). These preliminary results may provide some scientific support for the use of BJK for the anti-allergic treatment and inflammatory disorders through the inhibition of NO production.

Biological activities and chemical content of Sung Yod rice bran oil extracted by expression and soxhlet extraction methods.

BACKGROUND: Sung Yod rice is a red-violet pigmented rice and grown in the southern part of Thailand. Its rice bran oil has attracted the attention ofscientists who have described anti-oxidant properties ofsome ingredients in Sung Yod rice bran oil. Normally, extraction methods ofcommercial product from rice bran oil are by expression or soxhlet extraction with hexane. Thus, biological activities of Sung Yod rice bran oil related to health and chemical content ofrice bran oilfrom the two methods should be studied. OBJECTIVE: The objectives of this research were to investigate for biological activities and chemical content ofSung Yod rice bran oil obtainedfrom expression or soxhlet extraction method. MATERIAL AND METHOD: Biological activities such as cytotoxic, anti-inflammatory and antioxidant activities were investigated. Sulphorhodamine (SRB) assay was used to test cytotoxic activity against four human cancer cell lines: lung (COR-L23), cervical (HeLa), prostate (PC-3) and breast (MCF-7) and normal human lung cells (MRC-5). The inhibitory effect on lipopolysaccharide (LPS) induced nitric oxide (NO) production in RA W264. 7 cell lines was usedfor the determination of anti-inflammatory effect. DPPH, TEAC and FRAP assay were carried outfor antioxidant activity. Total phenolic compound was determined by Folin-Ciocalteu reagent. y-oryzanol and vitamin E content were determined by HPLC. Sung Yod rice bran oil was produced by expression method (EX) or by soxhlet extraction method using hexane (SXH-I). RESULTS: The percentage ofyield ofSung Yod rice bran oil by EX and SXH were 2.16 and 15.23 %w/w, respectively. Only EX showed the selective cytotoxicity against prostate cancer cells (PC-3), (IC50 = 52.06±1.60 µg/ml). It also exhibited high inhibitory effects on NO production (IC50 = 30.09 µg/ml). In contrast, SXH had no anti-inflammatory effect and cytotoxic activity against any of the cancer cells. EXshowed higher antioxidant activity determined using DPPH compared to SXH. It also showed higher amount of yoryzanol and vitamin E than that ofSXH (3.09±1.04 and 1.35±1.56 mg % w/w ofextracts, respectively). Yet, SXH exhibited higher antioxidant power determined by FRAP assay and higher total phenolic content compared with EX CONCLUSION: Sung Yod rice bran oil, produced by expression method, had better benefit for health regarding cytotoxicity against prostate cancer cells (PC-3), anti-inflammatory effect and antioxidant (using DPPH) than that produced by soxhlet method extracted with hexane.

Anti-allergic, anti-inflammatory and antioxidant activities of the different extracts of Thai traditional remedy called prabchompoothaweep for allergic rhinitis treatment.

BACKGROUND: Prabchompoothaweep remedy (PT) has long been used in Thai traditional medicine to treat allergic rhinitis and asthma. It is composed of 23 plants. It is on National herbal drug list of Thailand, but there is no reportfor anti-allergic, anti-inflammatory and antioxidant activities. OBJECTIVE: To investigate anti-allergic, anti-inflammatory and antioxidant activities of the crude extract from PTby different extraction method. MATERIAL AND METHOD: The method of extract used was maceration in 95% ethanol and 50% ethanol; the residue of these extracts were continued extracted by boiling water, they obtained PTE95, PTE50, PTR95 andPTR50, respectively. The other method of extraction was boiling and drying by lyophilizer that obtained PTW Five crude extracts were determined anti- allergic activity by the inhibition of ß-hexosaminidase release from RBL-2H3 cell lines, anti-inflammatory activity were determined by the inhibition ofnitric oxide (NO) production from RA W264. 7 cell lines induced by lipopolysaccharide (LPS) and antioxidant activity were tested by DPPH radical scavenging assay. RESULTS: PTE95 showed the most potent ofanti-allergic activity, anti-inflammatory activity and antioxidant activity (IC5 = 12.97, 22.51 and EC50 = 14.62 µg/ml, respectively). CONCLUSION: These results suggest that the method of extraction PT that showed the best anti-allergy, anti-inflamation and antioxidant activity was maceration in 95% ethanol.

An in vitro inhibitory effect on RAW 264.7 cells by anti-inflammatory compounds from Smilax corbularia Kunth.

BACKGROUND: Smilax corbularia is a Thai medicinal plant locally known as 'Hua-Khao-Yen Neua', which is used for treating inflammatory conditions. OBJECTIVE: To evaluate the anti-inflammatory effect of S. corbularia extracts and its isolated compounds by determination of inhibitory effects on lipopolysaccharide-stimulated PGE2 release, and TNF-alpha and NO production from RAW 264.7 cells. METHODS: The inhibitory effect of aqueous and ethanolic extracts of this plant were determined on LPS-induced NO production, TNF-alpha and PGE2 release in RAW 264.7 cells, as an in vitro indication of possible anti-inflammatory activity. The compounds from active extract were isolated by bioassay-guided fractionation. RESULTS: Only the ethanolic extract of this plant inhibited TNF-alpha and NO production, with IC50 values of 61.97, and 83.90 microg/ml respectively. Three flavonols, engeletin, astilbin and quercetin were isolated from the ethanolic extract. quercetin possessed the highest inhibitory effect on NO production with IC50 11.2 microg/ml (37.1 microM), whereas engeletin and astilbin had no activity (IC50 >100 microg/ml). All three flavonols possessed potent inhibition of PGE2 release with IC50 values of 14.4, 19.6 and 19.9 microg/ml (33.2, 43.5 and 65.8 microM) respectively. Quercetin also exhibited the highest inhibitory effect on TNF-alpha production (IC50 = 1.25 microg/ml or 4.14 microM), but engeletin and astilbin had no activity. CONCLUSION: This is the first report of isolated compounds from S. corbularia with potential anti-inflammatory effects, and the results support the use of this plant by Thai traditional doctors for treatment of inflammatory diseases.

Acetylcholinesterase inhibitory activity of Thai traditional nootropic remedy and its herbal ingredients.

The incidence of Alzheimer disease (AD) is increasing every year in accordance with the increasing of elderly population and could pose significant health problems in the future. The use of medicinal plants as an alternative prevention or even for a possible treatment of the AD is, therefore, becoming an interesting research issue. Acetylcholinesterase (AChE) inhibitors are well-known drugs commonly used in the treatment of AD. The aim of the present study was to screen for AChE inhibitory activity of the Thai traditional nootropic recipe and its herbal ingredients. The results showed that ethanolic extracts of four out of twenty-five herbs i.e. Stephania pierrei Diels. Kaempfera parviflora Wall. ex Baker, Stephania venosa (Blume) Spreng, Piper nigrum L at 0.1 mg/mL showed % AChE inhibition of 89, 64, 59, 50; the IC50 were 6, 21, 29, 30 microg/mL respectively. The other herbs as well as combination of the whole recipe had no synergistic inhibitory effect on AChE activity. However some plants revealed antioxidant activity. More research should have be performed on this local wisdom remedy to verify the uses in scientific term.

Antioxidant and nitric oxide inhibition activities of Thai medicinal plants.

Nineteen Thai medicinal plants used in Thai traditional medicine preparation to treat colds, asthma and fever were studied for their antioxidant and NO inhibitory activities. Three extracts were obtained from each plant. First extract obtained by macerating the plant part in 95% ethanol (Et) residue was boiled in water, where water extract (EW) was obtained. The third extract (HW) was obtained by boiling each plant in water similar to that of Thai traditional medicine practice. These extracts were tested for their antioxidant activity using DPPH assay, and anti-inflammatory activity by determination of inhibitory activity on lipopolysaccharide (LPS) induced nitric oxide (NO) production in RAW 264.7 cell lines using Griess reagent. Results indicated that Et, EW and HW of Syzygium aromaticum showed the highest antioxidant activity (EC50 = 6.56, 4.73 and 5.30 microg/ml, respectively). Et of Atractylodes lancea exhibited the most potent inhibitory activity on lipopolysaccharide (LPS) induced nitric oxide (NO) production in RAW 264.7 cells, with IC50 value of 9.70 microg/ml, followed by Et of Angelica sinensis and Cuminum cyminum (IC50 = 12.52 and 13.56 microg/ml, respectively) but water extract (EW, HW) of all plants were apparently inactive. These results of anti-inflammatory activity of these plants correspond with the traditional use for fever; cold, allergic-related diseases and inflammatory-related diseases.