Pawaret Panyajai's research while affiliated with Chiang Mai University and other places

Background and aims Curcuma aeruginosa, commonly known as “kha-min-dam” in Thai, holds significance in Asian traditional medicine due to its potential in treating various diseases, having properties such as anti-HIV, hepatoprotective, antimicrobial and anti-androgenic activities. This study explores the anticancer activity of C. aeruginosa essential oil (CAEO) and its nano-formulations. Methods CAEO obtained from hydrodistillation of C. aeruginosa fresh rhizomes was examined by gas chromatography mass spectroscopy. Cytotoxicity of CAEO was determined in leukaemic K562 and breast cancer MCF-7 cell lines using an MTT assay. Cell cycle analysis and cell apoptosis were determined by flow cytometry. Cell migration was studied through a wound-healing assay. Results Benzofuran (33.20%) emerged as the major compound of CAEO, followed by Germacrene B (19.12%) and Germacrone (13.60%). Two types of CAEO loaded nano-formulations, nanoemulsion (NE) and microemulsion (ME) were developed. The average droplet sizes of NE and ME were 13.8 ± 0.2 and 21.2 ± 0.2 nm, respectively. In a comparison with other essential oils from the fresh rhizomes of potential plants from the same family (Curcuma longa, Curcuma mangga and Zingiber officinale) on anticancer activity against K562 and MCF-7 cell lines, CAEO exhibited the highest cytotoxicity with IC50 of 13.43 ± 1.09 and 20.18 ± 1.20 µg/mL, respectively. Flow cytometry analysis revealed that CAEO significantly increased cell death, evidenced from the sub-G1 populations in the cell cycle assay and triggered apoptosis. Additionally, CAEO effectively inhibited cell migration in MCF-7 cells after incubation for 12 and 24 h. The developed NE and ME formulations significantly enhanced the cytotoxicity of CAEO against K562 cells with an IC50 of 45.30 ± 1.49 and 41.98 ± 0.96 µg/mL, respectively. Conclusion This study’s finding suggest that both nano-formulations, NE and ME, effectively facilitated the delivery of CAEO into cancer cells.

Hematopoietic stem cell transplantation (HSCT) is one method of lymphoma therapy. Peripheral blood stem cell (PBSC) separation, preservation, cell viability, and cell function before transplantation are important factors in the success rate of stem cell transplantation. This study aims to separate and estimate the efficiency of deep-freezing preservation in autologous PBSCs (CD34+/CD38-) from lymphoma patients undergoing stem cell transplantation. PBSCs were separated and collected by leukapheresis before being cryopreserved and stored in liquid nitrogen. The number of CD34+/CD45dim cells was investigated and compared with the subpopulation of CD34+/CD38- cells using conventional trypan blue exclusion method and 7-AAD before and after cryopreservation. Colony-forming units (CFUs) were determined to indirectly assess the viability and potency of the PBSCs. The result showed that CD34+/CD38– cells constituted 35.56% of total CD34+ cells and 0.05% of total nucleated cells (TNCs). After thawing, the number of CD34+/CD38- cells did not demonstrate significant differences compared with pre-storage. The percentage of CFU recovery was 94.74%. In this study, the storage process of deep-freezing cryopreservation demonstrated high-quality recovery of CD34+/CD38- cells from PBSCs for autologous hematopoietic stem cell transplantation in lymphoma patients. This result showed novel data about the preservation of CD34+/CD38- cells. Keywords: Lymphoma, Autologous peripheral blood stem cell, Blood cryopreservation, Bone marrow transplantation

Background and aims The purpose of this study was to investigate the biological properties of Kae-Lae (Maclura cochinchinensis (Lour.) Corner), a traditional medicinal plant used in Ayurvedic recipes in Thailand. To achieve this objective, heartwood samples were collected from 12 sources across Thailand. Fractional extracts (n-hexane, ethyl acetate, and ethanol) and the dominant compounds (morin, resveratrol, and quercetin) were examined for their abilities on cytotoxicity, antioxidant, anti-inflammation, and antileukaemic activity (Wilms’ tumour 1 protein was used as a well-known biomarker for leukaemic cell proliferation). Methods The study used MTT to assess cytotoxicity in leukaemic cells (K562, EoL-1, and KG-1a). Antioxidant activities were evaluated using ABTS, DPPH, and FRAP assays. The anti-inflammatory activity was investigated by detecting IL-2, TNF-α, and NO using appropriate detection kits. Wilms’ tumour 1 protein expression was measured by Western blotting to determine the anti-leukaemic activity. The inhibition of cell migration was also analyzed to confirm anticancer progression. Results Among the tested extract fraction, ethyl acetate No. 001 displayed strong cytotoxicity specifically in EoL-1 cells, while n-hexane No. 008 demonstrated this effect in three cell lines. Resveratrol, on the other hand, displayed cytotoxicity in all the tested cells. Additionally, the three major compounds, morin, resveratrol, and quercetin, exhibited significant antioxidant and anti-inflammatory properties. In particular, resveratrol demonstrated a noteworthy decreased Wilms’ tumour 1 protein expression and a reduction in cell proliferation across all cells. Moreover, ethyl acetate No. 001, morin, and resveratrol effectively inhibited MCF-7 cell migration. None of these compounds showed any impact on red blood cell haemolysis. Conclusion Based on these findings, it can be concluded that Kae-Lae has promising chemotherapeutic potential against leukaemic cells, with fractional extracts (ethyl acetate and n-hexane) and resveratrol exhibiting the most potent cytotoxic, antioxidant, anti-inflammatory, and anti-cell migration activities.

Golden cordyceps (Cordyceps militaris) is a mushroom of the genus Cordyceps. It has been used as a food supplement for both healthy and ill people. In this study, the antileukaemic cell proliferation activities of golden cordyceps extracts were examined and compared with standard cordycepin (CDCP) in EoL-1, U937, and KG-1a cells. Wilms’ tumour 1 (WT1) protein was used as a biomarker of leukaemic cell proliferation. The cytotoxicity of the extracts on leukaemic cells was determined using the MTT assay. Their inhibitory effects on WT1 protein expression and cell cycle progression of EoL-1 cells were investigated using Western blotting and flow cytometry, respectively. Induction of KG-1a cell differentiation (using CD11b as a marker) was determined using flow cytometry. The golden cordyceps extracts exhibited cytotoxic effects on leukaemic cells with the highest IC50 value of 16.5 ± 3.9 µg/mL, while there was no effect on normal blood cells. The expression levels of WT1 protein in EoL-1 cells were decreased after treatment with the extracts. Moreover, cell cycle progression and cell proliferation were inhibited. The levels of CD11b increased slightly following the treatment. All these findings confirm the antileukaemic proliferation activity of golden cordyceps.

Zingiber ottensii , is widely used in Asian traditional remedies for the treatment of many diseases. The present study explores anticancer activity of Z . ottensii essential oil (ZOEO) and its nanoformulations. ZOEO obtained from hydrodistillation of Z . ottensii fresh rhizomes was analysis using gas chromatography mass spectroscopy. Zerumbone (25.21%) was the major compound of ZOEO followed by sabinene (23.35%) and terpene-4-ol (15.97%). Four types of ZOEO loaded nanoformulations; nanoemulsion, microemulsion, nanoemulgels, and microemulgel, were developed. The average droplet size of the nanoemulsion and microemulsion was significantly smaller than that of the nanoemulgel and microemulgel. Comparison with other essential oils of plants of the same family on anticancer activity against A549, MCF-7, HeLa, and K562, ZOEO showed the highest cytotoxicity with IC 50 of 43.37±6.69, 9.77±1.61, 23.25±7.73, and 60.49±9.41 μg/mL, respectively. Investigation using flow cytometry showed that ZOEO significantly increased the sub-G1 populations (cell death) in cell cycle analysis and induced cell apoptosis by apoptotic analysis. The developed nanoformulations significantly enhanced cytotoxicity of ZOEO, particularly against MCF-7 with the IC 50 of 3.08±2.58, 0.74±0.45, 2.31±0.91, and 6.45±5.84 μg/mL, respectively. Among the four nanoformulations developed in the present study, nanoemulsion and microemulsion were superior to nanoemulgel and microemulgel in delivering ZOEO into cancer cells.

Acute myeloblastic leukemia (AML) is a disease with a high rate of relapse and drug resistance due to the remaining leukemic stem cells (LSCs). Therefore, LSCs are specific targets for the treatment of leukemia. CD123 is specifically expressed on LSCs and performs as a specific marker. Curcumin is the main active compound of a natural product with low toxicity for humans. It has been reported to inhibit leukemic cell growth. However, curcumin is practically insoluble in water and has low bioavailability. In this study, we aimed to formulate curcumin nanoparticles and conjugate with the anti-CD123 to overcome the low water solubility and improve the targeting of LSCs. The cytotoxicity of both curcumin-loaded PLGA/poloxamer nanoparticles (Cur-NPs) and anti-CD123-curcumin-loaded PLGA/poloxamer nanoparticles (anti-CD123-Cur-NPs) were examined in KG-1a cells. The results showed that Cur-NPs and Cur-NPs-CD123 exhibited cytotoxic effects on KG-1a cells with the IC50 values of 74.20 ± 6.71 and 41.45 ± 5.49 µM, respectively. Moreover, anti-CD123-Cur-NPs induced higher apoptosis than Cur-NPs. The higher uptake of anti-CD123-Cur-NPs in KG-1a cells was confirmed by using flow cytometry. In conclusion, the anti-CD123-Cur-NPs formulation improved curcumin’s bioavailability and specific targeting of LSCs, suggesting that it is a promising drug delivery system for improving the therapeutic efficacy against AML.

Kaffir lime (Citrus hystrix) is a plant member of family Rutaceae, and its leaves are commonly used in folk medicine. The present study explores antileukemic effects of the extracts and purified active compounds from the leaves. The antileukemic activity was investigated via inhibition of Wilms’ tumor 1 (WT1), which is a protein that involves in leukemic cell proliferation. In addition, the compounds were investigated for their effects on WT1 gene expression using real time RT-PCR and Western blotting. Cell cycle arrest and total cell number were investigated using flow cytometry and trypan blue exclusion method, respectively. The results demonstrated that the hexane fractionated extract had the greatest inhibitory effect on WT1 gene expression of many leukemic cell lines and significantly decreased WT1 protein levels of K562 cells (representative of the leukemic cells), in a dose- and time-dependent manner. Subfraction No. 9 (F9) after partial purification of hexane fractioned extract showed the highest suppression on WT1 protein and suppressed cell cycle at G2/M. The organic compounds were isolated from F9 and identified as phytol and lupeol. The bioassays confirmed antiproliferative activities of natural products phytol and lupeol. The results demonstrated anticancer activity of the isolated phytol and lupeol to decrease leukemic cell proliferation.

Leukemic cells remaining in the body is the main problem for cancer patients, and these cells are called Leukemic Stem Cells (LSCs). Many studies have revealed that the overexpression of the Wilms’ tumor 1 (WT1) protein is related to leukemogenesis. Curcuminoids (curcumin, demethoxycurcumin, and bisdemethoxycurcumin) from Thai turmeric (Curcuma longa Linn.) are the focus of this study because they have been previously been reported to show potent antileukemic activity. This study aims to investigate the cytotoxic effect of in-house curcuminoids on the human leukemic stem cell line (KG-1a) when compared to other leukemic cell lines (KG-1 and K562). MTT assays were used to determine the cytotoxicity of curcuminoids at various concentrations. Curcumin exhibited the strongest cytotoxic activity on KG-1a cells with IC50 values of 13.6 ± 1.9 µM. To determine the effect of curcuminoids on WT1 and CD34 protein expressions by Western blotting, KG-1a cells were treated with noncytotoxic concentrations (IC20 value). Bisdemethoxycurcumin showed the strongest suppression of WT1 and CD34 protein expressions with 73.3 ± 1.4 and 82.9 ± 2.0%, respectively. In summary, curcuminoids, especially bisdemethoxycurcumin, could inhibit WT1 and CD34 protein expressions. Thus, bisdemethoxycurcumin is a compound that calls for further studies of its process in the inhibition of WT1 and CD34 expressions in LSCs for the leukemia treatment.

Objective: The purpose of this study was to identify the chemical constituents in the dichloromethane extract by gas chromatography–mass spectrometry (GC–MS) analysis and evaluate the cytotoxic effects on leukemia cells of isolated compounds from Caesalpinia sappan Linn.Methods: Dichloromethane extract obtained from the heartwood of C. sappan was investigated by GC–MS and column chromatography. Cytotoxic effects on leukemia cells of the isolated compounds were examined using MTT assay.Results: The GC–MS analysis of dichloromethane extract from C. sappan revealed the presence of 14 compounds. Linoleic acid and β-sitosterol were found to be the major compounds presenting in 14% and 13%, respectively. The separation of the dichloromethane extract led to the isolation of brazilin (1) as a major compound, together with lupeol (2), and a mixture of β-sitosterol (3), and stigmasterol (4). Their structures were elucidated based on spectroscopic methods. Brazilin (1) showed a cytotoxic effect on human acute myeloid leukemia cell (KG1) and human acute myeloid leukemia stem cell (KG1a) with inhibitory concentration at 50% growth (IC50) values of 13.30 ± 0.49 and 12.24 ± 1.08 μg/ml, respectively. Conclusion: Many groups of phytochemical compounds in the dichloromethane extract of C. sappan were detected by GC–MS analysis. Some of them have been reported to possess various biological activities. Moreover, brazilin (1) isolated compound from C. sappan shows cytotoxicity on leukemia cells, which could be a potential anticancer property.