Concentrations Of Artesunate Research Articles

Molecular Interactions Identified by Two-Dimensional Analysis-Detailed Insight into the Molecular Interactions of the Antimalarial Artesunate with the Target Structure β-Hematin by Means of 2D Raman Correlation Spectroscopy.

A thorough understanding of the interaction of endoperoxide antimalarial agents with their biological target structures is of utmost importance for the tailored design of future efficient antimalarials. Detailed insights into molecular interactions between artesunate and β-hematin were derived with a combination of resonance Raman spectroscopy, two-dimensional correlation analysis, and density functional theory calculations. Resonance Raman spectroscopy with three distinct laser wavelengths enabled the specific excitation of different chromophore parts of β-hematin. The resonance Raman spectra of the artesunate-β-hematin complexes were thoroughly analyzed with the help of high-resolution and highly sensitive two-dimensional correlation spectroscopy. Spectral changes in the peak properties were found with increasing artesunate concentration. Changes in the low-frequency, morphology-sensitive Raman bands indicated a loss in crystallinity of the drug-target complexes. Differences in the high-wavenumber region were assigned to increased distortions of the planarity of the structure of the target molecule due to the appearance of various coexisting alkylation species. Evidence for the appearance of high-valent ferryl-oxo species could be observed with the help of differences in the peak properties of oxidation-state sensitive Raman modes. To support those findings, the relaxed ground-state structures of ten possible covalent mono- and di-meso(Cm)-alkylated hematin-dihydroartemisinyl complexes were calculated using density functional theory. A very good agreement with the experimental peak properties was achieved, and the out-of-plane displacements along the lowest-frequency normal coordinates were investigated by normal coordinate structural decomposition analysis. The strongest changes in all data were observed in vibrations with a high participation of Cm-parts of β-hematin.

Loss of Extrasynaptic Inhibitory Glycine Receptors in the Hippocampus of an AD Mouse Model Is Restored by Treatment with Artesunate.

Alzheimer's disease (AD) is characterized by synaptic failure and neuronal loss. Recently, we demonstrated that artemisinins restored the levels of key proteins of inhibitory GABAergic synapses in the hippocampus of APP/PS1 mice, a model of cerebral amyloidosis. In the present study, we analyzed the protein levels and subcellular localization of α2 and α3 subunits of GlyRs, indicated as the most abundant receptor subtypes in the mature hippocampus, in early and late stages of AD pathogenesis, and upon treatment with two different doses of artesunate (ARS). Immunofluorescence microscopy and Western blot analysis demonstrated that the protein levels of both α2 and α3 GlyRs are considerably reduced in the CA1 and the dentate gyrus of 12-month-old APP/PS1 mice when compared to WT mice. Notably, treatment with low-dose ARS affected GlyR expression in a subunit-specific way; the protein levels of α3 GlyR subunits were rescued to about WT levels, whereas that of α2 GlyRs were not affected significantly. Moreover, double labeling with a presynaptic marker indicated that the changes in GlyR α3 expression levels primarily involve extracellular GlyRs. Correspondingly, low concentrations of artesunate (≤1 µM) also increased the extrasynaptic GlyR cluster density in hAPPswe-transfected primary hippocampal neurons, whereas the number of GlyR clusters overlapping presynaptic VIAAT immunoreactivities remained unchanged. Thus, here we provide evidence that the protein levels and subcellular localization of α2 and α3 subunits of GlyRs show regional and temporal alterations in the hippocampus of APP/PS1 mice that can be modulated by the application of artesunate.

Pharmacokinetic and Bioanalytical Consideration of the Salivary Concentration of Artesunate and Dihydroartemisinin in Nigerian Volunteer Subjects: Using Spectrophotometric Methods

Pharmacokinetic and Bioanalytical Consideration of the Salivary Concentration of Artesunate and Dihydroartemisinin in Nigerian Volunteer Subjects: Using Spectrophotometric Methods

Assessment of quantitative and semi-quantitative biological test methods of artesunate in vitro.

Artesunate is the current most potent antimalarial drug widely used for the treatment of malaria. Considering the emergence of artemisinin resistance, several situations may require a simple method for artesunate quantification. We thus developed a quantitative and a semi-quantitative biological method for the determination of artesunate in liquid samples. The tests are based on the measurement of samples’ antimalarial activity on Plasmodium falciparum 3D7 using a modified SYBR Green I drug susceptibility test. For the quantitative test, we established a standard curve that resulted from a dose–response curve and evaluated its performances using controls samples. Whereas the linear regression analysis between artesunate concentration and antimalarial activity showed promising results (linearity range 1.5–24.6 ng/mL, r2 = 0.9373), we found that artesunate content of the controls was significantly overestimated (p = 0.0313). For the semi-quantitative test, we compared the antimalarial activities of samples collected during permeation studies of artesunate to that of a reference (artesunate IC50) by statistical analysis. We demonstrated that antimalarial activities of samples from permeation tests using a powder formulation of artesunate were greater than those of samples from tests using a solution formulation. Bioassays can be simple techniques to assess artesunate in liquid samples, particularly in resource-limited settings. Comparison with reference methods is still recommended when accurate drug quantification is required.

Artesunate-induced Cellular Effects Are Mediated by Specific EPH Receptors and Ephrin Ligands in Breast Carcinoma Cells

The aberrant regulation of erythropoietin-producing hepatocellular carcinoma (EPH) receptors and ephrin ligands has been implicated in breast carcinoma, and artesunate has been shown to have anticancer effects. The aim of this study was to characterize the involvement of EPH receptors and ephrin ligands in mediating artesunate (ART)-induced growth suppression of normal breast cells and breast carcinoma cell lines. The normal breast epithelial cells (MCF10A), non-invasive ductal breast carcinoma cells (MCF7), and invasive triple-negative breast carcinoma cells (MDA-MB-231) were grown in the absence or the presence of different concentrations of artesunate. The cells were counted, and total RNA was isolated. The abundance of transcripts corresponding to EPH receptors and ephrin ligands was determined by quantitative polymerase chain reaction. Cell viability was significantly reduced when cells were treated with artesunate, with MDA-MB-231 cells having the highest sensitivity. Artesunate had no significant effect on transcription of EPH/ephrins in MCF10A cells, but markedly increased EPHA8, EPHA10, EPHB6 and ephrin-A2 expression in MCF7 cells, and significantly increased EPHA3 and EPHA10 expression while reducing that of EPHA7 and ephrin-A3 in MDA-MB-231 cells. The relative changes in artesunate-treated MCF7 and MDA-MB-231 cells as compared to similarly treated MCF10A cells allow us to implicate combinatorial expression and receptor interactions for EPH receptor-mediated signal transduction that converges into pathways responsible for cell growth, proliferation, and apoptosis. Specifically, the alterations in EPHA7, EPHA8, EPHA10 and EPHB6 transcripts appear to be important participants in artesunate-mediated cellular effects.

In vitro efficacy of artemisinin-based treatments against SARS-CoV-2

Effective and affordable treatments for patients suffering from coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), are needed. We report in vitro efficacy of Artemisia annua extracts as well as artemisinin, artesunate, and artemether against SARS-CoV-2. The latter two are approved active pharmaceutical ingredients of anti-malarial drugs. Concentration–response antiviral treatment assays, based on immunostaining of SARS-CoV-2 spike glycoprotein, revealed that treatment with all studied extracts and compounds inhibited SARS-CoV-2 infection of VeroE6 cells, human hepatoma Huh7.5 cells and human lung cancer A549-hACE2 cells, without obvious influence of the cell type on antiviral efficacy. In treatment assays, artesunate proved most potent (range of 50% effective concentrations (EC50) in different cell types: 7–12 µg/mL), followed by artemether (53–98 µg/mL), A. annua extracts (83–260 µg/mL) and artemisinin (151 to at least 208 µg/mL). The selectivity indices (SI), calculated based on treatment and cell viability assays, were mostly below 10 (range 2 to 54), suggesting a small therapeutic window. Time-of-addition experiments in A549-hACE2 cells revealed that artesunate targeted SARS-CoV-2 at the post-entry level. Peak plasma concentrations of artesunate exceeding EC50 values can be achieved. Clinical studies are required to further evaluate the utility of these compounds as COVID-19 treatment.

Pharmacokinetic Study of Rectal Artesunate in Children with Severe Malaria in Africa.

When severe malaria is suspected in children, the WHO recommends pretreatment with a single rectal dose of artesunate before referral to an appropriate facility. This was an individually randomized, open-label, 2-arm, crossover clinical trial in 82 Congolese children with severe falciparum malaria to characterize the pharmacokinetics of rectal artesunate. At admission, children received a single dose of rectal artesunate (10 mg/kg of body weight) followed 12 h later by intravenous artesunate (2.4 mg/kg) or the reverse order. All children also received standard doses of intravenous quinine. Artesunate and dihydroartemisinin were measured at 11 fixed intervals, following 0- and 12-h drug administrations. Clinical, laboratory, and parasitological parameters were measured. After rectal artesunate, artesunate and dihydroartemisinin showed large interindividual variability (peak concentrations of dihydroartemisinin ranged from 5.63 to 8,090 nM). The majority of patients, however, reached previously suggested in vivo IC50 and IC90 values (98.7% and 92.5%, respectively) of combined concentrations of artesunate and dihydroartemisinin between 15 and 30 min after drug administration. The median (interquartile range [IQR]) time above IC50 and IC90 was 5.68 h (2.90 to 6.08) and 2.74 h (1.52 to 3.75), respectively. The absolute rectal bioavailability (IQR) was 25.6% (11.7 to 54.5) for artesunate and 19.8% (10.3 to 35.3) for dihydroartemisinin. The initial 12-h parasite reduction ratio was comparable between rectal and intravenous artesunate: median (IQR), 84.3% (50.0 to 95.4) versus 69.2% (45.7 to 93.6), respectively (P = 0.49). Despite large interindividual variability, rectal artesunate can initiate and sustain rapid parasiticidal activity in most children with severe falciparum malaria while they are transferred to a facility where parenteral artesunate is available. (This study has been registered at ClinicalTrials.gov under identifier NCT02492178.).

Preclinical Evaluation of Artesunate as an Antineoplastic Agent in Ovarian Cancer Treatment.

Background: Ovarian cancer is the deadliest gynecologic malignancy despite current first-line treatment with a platinum and taxane doublet. Artesunate has broad antineoplastic properties but has not been investigated in combination with carboplatin and paclitaxel for ovarian cancer treatment. Methods: Standard cell culture technique with commercially available ovarian cancer cell lines were utilized in cell viability, DNA damage, and cell cycle progression assays to qualify and quantify artesunate treatment effects. Additionally, the sequence of administering artesunate in combination with paclitaxel and carboplatin was determined. The activity of artesunate was also assessed in 3D organoid models of primary ovarian cancer and RNAseq analysis was utilized to identify genes and the associated genetic pathways that were differentially regulated in artesunate resistant organoid models compared to organoids that were sensitive to artesunate. Results: Artesunate treatment reduces cell viability in 2D and 3D ovarian cancer cell models. Clinically relevant concentrations of artesunate induce G1 arrest, but do not induce DNA damage. Pathways related to cell cycle progression, specifically G1/S transition, are upregulated in ovarian organoid models that are innately more resistant to artesunate compared to more sensitive models. Depending on the sequence of administration, the addition of artesunate to carboplatin and paclitaxel improves their effectiveness. Conclusions: Artesunate has preclinical activity in ovarian cancer that merits further investigation to treat ovarian cancer.

Artesunate, as a HSP70 ATPase activity inhibitor, induces apoptosis in breast cancer cells

The present study aims to evaluate the inhibitory effects of artesunate (a semi-synthetic derivative of artemisinin) on HSP70 and Bcl-2 expression in two breast cancer cell lines, 4T1 and MCF-7. In addition, to determine in vitro inhibitory effect of artesunate against the ATPase activity of purified recombinant HSP70, it was tested in a carbonic anhydrase refolding assay with purified HSP70. Our results demonstrated that the artesunate not only induced apoptosis but also lead to the inhibition of HSP70 ATPase activity the in vitro (P < 0.001). The extent of HSP70 refolding inhibition increased with increasing μM concentrations of artesunate. Incubation of HSP70 with 50 μM artesunate showed significant inhibition of refolding activity by 38%. The IC50 values of artesunate for 4T1 cells, were lower than MCF-7 cells, indicating the higher sensitivity of the triple-negative phenotype. Furthermore, artesunate significantly down-regulated the expression of Bcl-2 and HSP70 while enhancing the expression of cleaved caspase-9 in MCF-7 and 4T1 cells. It also induced caspase-9 activity at 18 h in a dose-dependent manner in two breast cancer cell lines. Generally, our results show that the artesunate induces caspase-dependent apoptosis through the inhibition of HSP70 expression.

Development of colorimetric method for the assay of artesunate using 4-nitrobenzaldehyde

Purpose: Artesunate is one of the artemisinin derivatives that is commonly used in combination with some anti-malarials as artemisinin-based combination therapies. High performance liquid chromatographic technique is the required standard assay method for artesunate, but this is not readily available or affordable in most developing countries. This study was carried out to develop a simple, accurate and cost-effective assay method for quantitative determination of artesunate. Methods: The method is an indirect colorimetric assay, which was developed from the formation of yellow coloured product due to the reaction between acid decomposed product of artesunate with 4-nitrobenzaldehyde. Results: The wavelength of maximum absorption for the yellow coloured product was 474 nm. The Beer’s law was obeyed at the range of 20-100 µg/ml artesunate concentration with a linear coefficient of 0.9996. The molar absorptivity was 2.3183×103 mol-1 cm-1. The limit of detection and quantification were 0.0753 µg/ml and 0.2283 µg/ml, respectively. The method required water as diluent. The result obtained from recovery study confirmed that there was no interference from pharmaceutical excipients. Five brands of artesunate tablets were assayed using the developed method. Conclusion: The results compared favourably well with those obtained using the official method described in the international pharmacopoeia. The developed method is useful for the quantitative determination of artesunate in tablets and raw pharmaceutical material. Keywords: Artemisinin, quantitative determination, in-direct colorimetric assay, molar absorptivity

Effect of artesunate and relation with TGF-\u03b21 and SMAD3 signaling on experimental hypertrophic scar model in rabbit ear

Artesunate (ART) is the derivative of artemisinin isolated from the traditional Chinese medicine qinghao. Although several studies reported the efficiency of artesunate in the treatment of malaria, inhibiting fibroblasts and collagen synthesis, the association between artesunate and scar formation is unclear. The research was designed to study the significance of artesunate (ART) on the expression of transforming growth factor (TGF-β1) and small mother against decapentaplegic (SMAD3) in rabbit's ear hypertrophic scar model. Twenty-four New Zealand white rabbits were randomly divided into six groups: control group, matrix group, low-concentration artesunate group (0.48%), medium-concentration artesunate group (0.96%), high-concentration artesunate group (1.92%) and silicone gel group. Punched defects were established on each rabbit’s ear which resulted in a hypertrophic scar. On the 28th day, topical artesunate creams were applied twice a day except on the control group. On the 56th day, scar samples were collected for histopathology and immunoassay. Hematoxylin and eosin staining, Van Gieson staining, immunohistochemistry and Western blot analysis were done. Amongst the six groups, findings showed that the medium-concentration artesunate group (0.92%) efficiently decreased hypertrophic scar formation and significantly reduced fibroblasts and collagen synthesis. The results had also shown a decrease in the expression of transforming growth factor (TGF-β1) and declined small signal mother against decapentaplegic (Smad3). The overall study shows efficacy and mechanism of artesunate. It concluded that the medium concentration of artesunate (0.92%) could be an effective therapeutic agent for hypertrophic scars.

Safety assessment of MEFAS: an innovative hybrid salt of mefloquine and artesunate for malaria treatment

Malaria is a global public health problem that causes approximately 445 000 deaths annually worldwide, especially in underdeveloped countries. Because of the high prevalence and mortality of the disease, new and less toxic therapeutic agents need to be developed, such as MEFAS, a low-cost hybrid salt that consists of artesunate and mefloquine. However, the efficacy of MEFAS has been systematically demonstrated, its safety requires further investigation. This study investigated the acute toxicity of MEFAS and its precursors, artesunate, and mefloquine. A total of 42 female Swiss mice were divided into seven groups (n = 6/group) that were treated orally by gavage with vehicle (filtered water, negative control), MEFAS (50, 500, and 1000 mg/kg), and 1:1 concentrations of artesunate + mefloquine (50, 500, and 1000 mg/kg). Clinical signs of toxicity were observed for 14 d after treatment. On day 15, the animals were weighed, deeply anesthetized with isoflurane, and euthanized for subsequent collection of the liver, spleen, and kidneys. The relative organ weights were determined, followed by histopathological analysis. Artesunate + mefloquine produced toxic effects compared with the negative control group, reflected by changes in clinical signs, relative organ weights, and histopathological alterations. In MEFAS-treated animals, no changes were observed compared with the negative control group. These findings demonstrate that MEFAS is safer than artesunate + mefloquine after acute administration in mice.

Effects of artesunate combined with bortezomib on apoptosis and autophagy of acute myeloid leukemia cells in vitro and its mechanism

目的探讨青蒿琥酯联合硼替佐米对急性髓系白血病（AML）细胞株MV4-11细胞增殖、凋亡及自噬的影响及其作用机制。方法MTT法检测青蒿琥酯、硼替佐米单用及两药联合对MV4-11细胞的增殖抑制作用，流式细胞术检测细胞凋亡情况，Western blot法检测细胞内Bcl-2家族蛋白（Bcl-2、Mcl-1、Bim、Bax）、cleaved-Caspase-3及自噬相关蛋白LC3B的表达。结果青蒿琥酯呈浓度和时间依赖性抑制MV4-11细胞增殖，作用48 h的IC50为1.44 µg/ml。硼替佐米呈浓度依赖性抑制MV4-11细胞增殖，作用48 h的IC50为8.97 nmol/L。青蒿琥酯（0.75、1.0 µg/ml）与硼替佐米（6、8 nmol/L）联合作用48 h，对MV4-11细胞的增殖抑制率均高于单药组（P值均<0.05），两药相互作用的协同指数（CI）均<1。1.5 µg/ml青蒿琥酯作用MV4-11细胞48 h的凋亡率为（15.27±2.18）%，8 nmol/L硼替佐米作用MV4-11细胞48 h的凋亡率为（19.85±3.23）%，两药联合作用后凋亡率增高至（81.67±5.96）%，显著高于单药组（P值均<0.05）。两药联合作用MV4-11细胞24 h后细胞内cleaved-Caspase-3、促凋亡蛋白Bim及自噬相关蛋白LC3B表达上调，抗凋亡蛋白Bcl-2表达下调。结论青蒿琥酯联合硼替佐米具有协同抑制MV4-11细胞增殖和诱导凋亡及促进自噬的效应，作用机制可能与Bcl-2家族蛋白表达的改变有关。

The Antifungal Activity of Artesunate toward Candida albicans: Two Opposite Activities

The artemisinin and its derivatives antifungal activity continue to be an interesting research object, with the potential shown to be developed as an antifungal compound. Artesunate, one of the artemisinin derivatives known to have antifungal activity against various pathogenic fungi, including Candida albicans. This study aims to determine the effect of artesunate on antifungal activity toward C. albicans in vitro at concentrations below 1 mgmL-1. The method used is yeast-plate count, with a parameter of observation were the number of C. albicans colonies viable after exposure with artesunate for five days. The concentration of artesunate used was divided into six groups, which were 10; 1; 10-1; 10-2; 10-3; and 10-4mgmL-1. Compared to control, a significant decrease in colony counts was only shown at the highest concentration of 10 mgmL-1. Interestingly, at the lowest concentration of 10-4mgmL-1, it showed an increase in a number of colonies almost twice of the blank. These results suggest that while at higher concentration of artesunate may inhibit the growth of C. albicans, a lower concentration of artesunate may stimulate their growth.

Artesunate Suppresses Cell Proliferation of THP-1 Cells by Inducing Apoptosis

To investigate whether Artesunate(ART) can inhibit the proliferation of THP-1 cells and to explore the potential mechanism of its anti-leukemia effect. THP-1 cells were treated with 5 concentrations of Artesunate for 24 h, 48 h or 72 h. The viability of cells was detected with CCK-8 assay, apoptosis was assessed by using flow cytometry, and the STAT3, Caspase3 and Caspase8 protein levels were measured with Western blot . Compared with the control group, ART significantly inhibited the proliferation of THP-1 cells in a dose-dependent manner (r=0.9829, P<0.05). ART also increased the apoptosis of THP-1 cells. The results of Western blot showed that after treated with ART, the STAT3 protein expression in THP-1 cells was significantly down-regulated (P<0.01), and the expressions of Caspase3, cleaved Caspase3 and Caspase8 proteins were up-regulated(P<0.01). Artesunate can inhibit the proliferation of THP-1 cells, which may relate with the down-regulation of STAT3 expression and the activation of Capase3 and Caspase8.

The therapeutic effect of artesunate on rosacea through the inhibition of the JAK/STAT signaling pathway.

Acne rosacea is a type of chronic dermatosis with the characteristics of erubescence, angiotelectasis and pustule formation. However, current treatment methods are limited due to the side effects. Artesunate demonstrated a promising therapeutic efficacy with a high safety margin. HaCaT cells were treated with antibacterial peptide LL‑37 to simulate rosacea caused by Demodex folliculorum (D. folliculorum) infection. Cell Counting kit 8 and flow cytometry assays were performed to measure cellular proliferation, apoptosis, the stage of the cell cycle and reactive oxygen species generation in order to determine the level of cell damage. Then the damaged cells were treated with different concentrations of artesunate and doxycycline to determine the therapeutic effect of artesunate. Pro‑inflammatory cytokines tumor necrosis factor‑α (TNF‑α), interleukin (IL)‑6, IL‑8 and C‑C motif chemokine 2 (MCP‑1) were measured using an ELISA, while western blotting was used to detect the expression of Janus kinase 2 (JAK2) and signal transducer and transcription activator (STAT3). As a result, LL‑37 treated HaCaT cells decreased in cell viability, had an increased apoptotic rate and cell cycle arrest, indicating that cell damage caused by rosacea was simulated. In addition, upregulated concentrations of the pro‑inflammatory cytokines TNF‑α, IL‑6, IL‑8 and MCP‑1 were attenuated in the artesunate group in a dose‑dependent fashion, indicating the therapeutic effect of artesunate. Furthermore, higher concentrations of artesunate exhibited an improved effect compared with the doxycycline group. In addition, increased expression levels of JAK2 and STAT3 following treatment with LL‑37 suggested that rosacea caused by D. folliculorum infection may lead to inflammation through the JAK/STAT signaling pathway. In conclusion, the potential mechanism by which damage occurs in rosacea was revealed and a promising therapeutic method against rosacea was demonstrated.

Role of artesunate in TGF‑β1‑induced renal tubular epithelial‑mesenchymal transdifferentiation in NRK‑52E cells.

The implications of epithelial-mesenchymal transdifferentiation (EMT) have extended beyond the confines of renal fibrosis to renal tubulointerstitial fibrosis. It has been proposed that EMT may be one of the mechanisms involved in the pathogenesis of renal fibrosis. However, the underlying mechanisms remain unknown. Transforming growth factor (TGF)-β1 is considered to be an important cytokine which regulates the transdifferentiation of tubular epithelial cells into myofibroblasts in renal tubulointerstitial fibrosis. In the present study, normal rat kidney tubular epithelial cells (NRK-52E) were treated for 48 h with TGF-β1 (5 ng/ml) and different concentrations of artesunate (ART; 0.01, 0.1 and 1 µg/ml). Western blotting, reverse transcription-semi quantitative polymerase chain reaction analysis and immunofluorescence staining were used to evaluate the expression of bone morphogenetic protein (BMP)-7, uterine sensitization-associated gene (USAG)-1, E-cadherin, α-smooth muscle actin (α-SMA) and extracellular matrix collagen type I (Col I) mRNA. ART was able to attenuate renal injury in a unilateral ureteral obstruction model. However, its anti-fibrotic effect remains to be elucidated. In the present study, it was observed that ART was able to ameliorate the TGF-β1-induced alterations in cellular morphology. In addition, ART inhibited the TGF-β1-induced USAG-1 increase and the decrease in BMP-7. Treatment with ART markedly attenuated the TGF-β1-induced upregulation of α-SMA and downregulation of E-cadherin. Additionally, ART was able to significantly attenuate the deposition of interstitial collagens, including Col I. The results of the present study further verified the therapeutic efficacy of ART in TGF-β1-induced renal interstitial fibrosis. These findings indicated that ART may hold the potential to prevent chronic kidney diseases via the suppression of USAG-1 expression or by increasing BMP-7 expression.

Inhibitory Effect of Artesunate on Growth and Apoptosis of Gastric Cancer Cells

The present study aimed to explore the inhibitory effect of artesunate on the growth and apoptosis of gastric cancer cells, in order to search for high effective and low toxic anti-gastric cancer drugs. Flow cytometry was used to detect the CDC25A protein expression in different gastric diseases. After the treatment of different concentrations of artesunate (0, 30, 60, 120μmol/l) on the gastric cancer cell line SGC-7901cells for 24h, 0μmol/l Art was instead of normal saline as control group, the expression of CDC25A, Bcl-2, Bax, Caspase-3 protein, mitochondrial membrane potential, cell apoptosis and cell cycle were detected by flow cytometry. The CDC25A protein expression in gastric adenocarcinoma was significantly higher than that in normal gastric tissues. After the treatment of different concentrations of Art, the apoptosis rate of SGC-7901cells in Art groups was significantly higher than that in control group. The proliferation index and CDC25A protein of SGC-7901cells in Art groups was significantly lower than that in control group. Compared with control group, the Bcl-2 protein and mitochondrial membrane potential was significantly lower but the Bax and Caspase-3 protein expression level was significantly higher. The high expression of CDC25A protein in gastric adenocarcinoma is involved in the occurrence and development of gastric adenocarcinoma. Artesunate can inhibit the growth of gastric adenocarcinoma cells SGC-7901 and induce the cell apoptosis, the mechanism may be related to the regulation of CDC25A, Bcl-2, Bax, Caspase-3 and mitochondrial membrane potential in SGC-7901cells.

Induction of Apoptosis in Human Breast Cancer MCF-7 Cells by a Semi-Synthetic Derivative of Artemisinin: A Caspase-Related Mechanism.

Background: Artesunate has recently been used in some pharmacological preparation to induce tumor cell apoptosis. The drug is a semi-synthetic derivative of artemisinin, traditionally used for its antimalarial. However, up to now, its anticancer mechanism against different types of tumors is not known. Objectives: The most important purposes of the present research was firstly investigating induction of apoptosis on human breast cancer MCF-7 cells by the drug and, in the second place, introducing its possible mechanism of action. Materials and Methods: The MTT assay was used to investigate the inhibitory effect of artesunate on growth of breast cancer MCF-7 cells. For this aim, different concentrations of artesunate were used to treat the cells and flow cytometry assay was done followed by annexin V-FITC/PI staining. The activities of caspase-3, -8 and -9 were then determined by relative assay kits. Results: Based on the results from MTT assay, it was found that artesunate could significantly inhibit the growth of MCF-7 cells in a dose- and time-dependent manner. On the other hand, the flow cytometry findings showed that the anti-proliferative activity of artesunate on MCF-7 cells is due to apoptosis. Besides, caspase colorimetric assays revealed a significant rise in cellular levels of the initiators (caspase-8 and -9) and effector (caspase-3) in the cells treated by artesunate. Conclusions: According to our results, it could be concluded that artesunate could inhibit the growth of MCF-7 breast cancer cells through induction of apoptosis by intrinsic and extrinsic caspase-dependent pathways. Therefore, we claim that artesunate could be introduced as a suitable candidate for the treatment of the breast cancer.

In vitro inhibitory effect of artesunate on the vascular endothelial growth factor secreted by mononuclear cells in hand-foot-and-mouth disease children complicated with encephalitis

Objective To investigate the dynamics of white blood cell count (WBC), prealbumin (PA), high sensitive C-reactive protein (hs-CRP), and vascular endothelial growth factor (VEGF) in hand-foot-and-mouth disease (HFMD) complicated with or without encephalitis and in vitro inhibitory effects of artesunate on VEGF secretion of mononuclear cells from HFMD patients. Methods Peripheral blood mononuclear cells from healthy control group, HFMD group and HFMD combined with encephalitis group were isolated by Ficoll density gradient centrifugation and treated with different concentrations of artesunate (25, 50, 100 mg/L). The expression of VEGF in the supernatant was examined by ELISA double antibody sandwich method. The levels of WBC, PA and hs-CRP in the three groups were also detected. Multiple samples were compared by one-way analysis of variance. Multiple comparison was performed by Dunnett T3 test. Correlation of two variables was analyzed by Spearman correlation test. Results There were significant differences in the levels of WBC, hs-CRP, PA and VEGF between the HFMD combined with or without encephalitis group and health control group (F=172.69, 366.02, 166.32 and 5 941.89, respectively, all P<0.01). There were significant differences in the levels of VEGF secreted by mononuclear cells treated with three different concentrations of artesunate between the HFMD combined with or without encephalitis groups and health control group (F=194.265 and 4 750.69, respectively, both P<0.01). Correlation analysis showed that the VEGF level secretion by mononuclear cells from HFMD patients with or without encephalitis were both negatively correlated with different concentrations of artesunate (r=-0.903 and -0.969, both P<0.01). Conclusions Compared with HFMD without encephalitis, the secretion of VEGF by mononuclear cells, WBC and hs-CRP levels in HFMD complicated with encephalitis group all increase and PA decrease significantly. Artesunate can inhibit the secretion of VEGF by mononuclear cells of patients with hand-foot-and-mouth disease in a dose-dependent manner in vitro. Key words: Hand, foot and mouth disease; Encephalitis; Artesunate; Vascular endothelial growth factors