Effect of Hispidulin on Proliferation and Apoptosis of Leukemia K562 Cells by Up-Regulating IL-37 / 中国实验血液学杂志

OBJECTIVE@#To investigate the effect and underlying mechanism of hispidulin on the proliferation and apoptosis of leukemia K562 cells.@*METHODS@#K562 cells were cultured in vitro and treated with 0, 5, 25 or 100 μmol/L hispidulin for 24 h. Cell proliferation and apoptosis were detected by CCK-8 and flow cytometry, respectively. Western blot was used to assess the expression of Bax, Bcl-2 and interleukin (IL)-37 proteins. Bone marrow mononuclear cells were extracted from 17 chronic myeloid leukemia patients and 21 healthy individuals by Ficoll-Hypaque density gradient method, and the expression of IL-37 protein was measured by Western blot. K562 cells with IL-37 overexpression or knockdown were constructed, and then treated with 0 or 100 μmol/L hispidulin for 24 h. Cell proliferation, apoptosis and protein expression of Bax and Bcl-2 were determined in the same way as above.@*RESULTS@#After K562 cells were treated with hispidulin, the cell inhibition rate, apoptosis rate, and the protein expression of Bax and IL-37 were significantly increased (P <0.05), but the cell proliferation and expression of Bcl-2 protein were decreased (P <0.05). The expression of IL-37 protein in bone marrow mononuclear cells of the leukemia patient was 0.24±0.03, which was significantly lower than 0.91±0.05 of healthy controls (P <0.05). Overexpression of IL-37 significantly promoted inhibition rate, apoptosis rate, and expression of Bax protein in K562 cells (P <0.05), but suppressed the expression of Bcl-2 protein (P <0.05). In addition, knockdown of IL-37 could reverse the effects of hispidulin on proliferation and apoptosis of K562 cells.@*CONCLUSION@#Hispidulin inhibits the proliferation and induces apoptosis of leukemia K562 cells, which may be related to the up-regulation of IL-37 protein in cells.

Preparation of hispidulin chitosan microsphere and its antiproliferation effect / 中国中药杂志

Hispidulin(HPDL) chitosan microspheres were prepared in this study to deliver HPDL to the lesion sitevia intravenous injection, and further evaluate their anticancer effects in vitro and the growth inhibition effect on A549 cells spheroids. HPDL chitosan microspheres were prepared by emulsion crosslinking method with chitosan as a drug carrier and the amount of HPDL was determined by high performance liquid chromatography (HPLC). The morphology of microspheres was observed under laser scanning confocal microscope. Additionally, the drug release amount of targeting microspheres was detected by dialysis method. Furthermore, the anti-proliferative effects against A549 lung cancer cells were tested by sulforhodamine B (SRB) method, and the effects of HPDL chitosan micrpsphereson early apoptosis of A549 cellswere determined by flow cytometry. A549 cells tumor spheroids were developed in vitro and then HPDL chitosan microspheres were added. On the 0, 1, 3, 7 d after adding the drugs, the inverted microscope was used to observe the mythologicaland volume changes of A549 cells spheroids. The encapsulation efficiency of HPDL chitosan microspheres was （75.32±0.52）%, and the drug loading amount was （7.76±0.67）%. Meanwhile, the microspheres were round shaped andhad smooth surface. The HPDL chitosan microspheres exhibited stronger inhibitory effects on A549 lung cancer cells. The results of flow cytometry indicated that, the early apoptosis rate of lung cancer A549 cells was （37.0±0.75）% at 24 h cells culture after drug administration. The volume of tumor spheroid was significantly inhibited, which had been shrunk by (50.09±11.06)% after the treatment by drug-loaded microsphere at day 7 as compared with blank group; meanwhile, the cells surface were obviously lysed. The preparation method in this research was simple and practicable, and the microspheres prepared with this method were round and smooth, with high encapsulation efficiency, which can significantly inhibit proliferation of lung adenocarcinoma A549 cells and induce cell apoptosis, and at the same time can cause lysisand death of A549 cell tumor spheroid.

Preparation of Hispidulin Liposomes and Its Antitumor Effect in Vitro / 中国药学杂志

OBJECTIVE: To study the preparing METHODS of hispidulin liposomes and evaluate their antitumor effects in vitro. METHODS: Hispidulin liposomes were prepared by thin film dispersion method, and the formulation was further optimized. At the same time, the liposomes particle size and Zeta potential were characterized separately. The antiproliferative effect of the liposomes on A549 cells was investigated by sulforhodamine B(SRB) method. RESULTS: According to the optimal prescription, the encapsulation efficiency of hispidulin liposomes was (87.06±0.67)%. Additionally, the average particle size was (104.83±1.40) nm and the Zeta potential was (-4.61±0.32) mV. Furthermore, the hispidulin liposomes exhibited the strongest inhibiting effect on A549 cells in vitro with comparison with control groups. CONCLUSION: The preparation method builtin our research is easy. And the liposomes are well distributed with high encapsulation efficiency. Meanwhile, the hispidulin liposomes exhibit the most significant antitumor effects in vitro.

Anti-tumor Activity of Saussurea laniceps against Pancreas Adenocarcinoma

The purpose of this study was to confirm the anti-tumor activity of an ethanol extract of Saussurea laniceps against pancreatic tumor and to isolate the active compound from S.laniceps extract. Treatment with S.laniceps extract and hispidulin inhibited proliferation of pancreatic cell lines, such as Capan-1, Capan-2, Panc-1 and S2-013 in a dose-dependent manner using the hollow fiber assay. Hispidulin showed typical hallmarks of apoptotic cell death a significant anti-tumor activity on Capan-2 cells at a dose of 100 mg/kg and 200 mg/kg. S.laniceps has potential cytotoxic and apoptotic effects on human pancreatic carcinoma cells. Its mechanism of action might be associated with the apoptotic cell death through DNA fragmentation.

Chemical constituents from Scutellaria barbata / 中草药

Objective: To investigate the constituents in the aerial parts of Scutellaria barbata. Methods: The isolation and purification of the compounds were performed by AB-8 macroporous adsorption resin, silica gel, polyamide and Sephadex LH-20 column chromatography, and their structures were determined on physicochemical characters and spectroscopic data. Results: Fourteen compounds were separated and elucidated as hispidulin-7-O-β-D-methylgluzcuronide (1), apigenin (2), scutellarin (3), scutellarein (4), luteolin (5), scutellarein-7-O-β-D-glucuronide methyl ester (6), isoscutellarein-8-O-β-D-glucuronide-6″-methyl ester (7), apigenin-7-O-β-D-glucuronide-6″-methyl ester (8), 4'-hydroxywogonin (9), 4',5-dihydroxy-3',5',6,7-tetramethoxyflavone (10), isoscutellarein (11), 6-hydroxyluteolin (12), 5-hydroxy-6,7,3',4'-tetramethoxyflavone (13), salvigenin (14). Conclusion: Compound 7 is isolated from Lamiaceae for the first time, compounds 1 and 13 are for the first time isolated from the genus Scutellaria, and compounds 6 and 12 are for the first time obtained from S. barbata.

Effects of natural flavones on membrane properties and citotoxicity of HeLa cells / Efeitos de flavonas naturais em propriedades de membranas e em citotoxicidade de células HeLa

The aim of this study was to determine whether eupafolin and hispidulin, flavones extracted from Eupatorium littorale Cabrera, Asteraceae, have the ability to change properties of biological membranes and promote cytotoxic effects. Eupafolin (50-200 µM) decreased approximately 30 percent the rate and total amplitude of valinomycin induced swelling and 60-100 percent the energy-dependent mitochondrial swelling. Moreover, eupafolin (200 µM) reduced 35 percent the mitochondrial permeability transition, and hispidulin did not change this parameter in any of the doses tested. The evaluation of phase transition of DMPC liposomes with the probe DPH demonstrated that hispidulin and eupafolin affect gel and fluid phase. With mitochondrial membrane as model, hispidulin increased the polarization of fluorescence when used DPH-PA probe. Eupafolin and hispidulin (100 µM) promoted a reduction of 40 percent in cellular viability of HeLa cells in 24 h. Our results suggest that eupafolin and hispidulin have cytotoxic effects that can be explained, in part, by alterations promoted on biological membranes properties and mitochondrial bioenergetics.

O objetivo deste estudo foi avaliar se eupafolina e hispidulina, flavonas extraídas do Eupatorium littorale Cabrera, Asteraceae, possuíam a capacidade de alterar propriedades das membranas biológicas e promover efeitos citotóxicos. Eupafolina (50-200 µM) reduziu em aproximadamente 30 por cento a velocidade e amplitude do inchamento mitocondrial induzido por valinomicina e 60-100 por cento o inchamento mitocondrial dependente de substrato. Além disso, eupafolina na dose de 200 µM reduziu a transição de permeabilidade mitocondrial em 35 por cento entretanto, a hispidulina não alterou este parâmetro em todas as doses testadas. A avaliação da transição de fase dos lipossomas de DMPC com a sonda DPH demonstrou que ambas as flavonas afetam a fase gel e fluida. Quando lipossomas de membranas mitocondriais e a sonda DPH-PA foram utilizados, houve aumento da polarização de fluorescência promovido pela hispidulina. Eupafolina e hispidulina, na dose de 100 µM, promoveram 40 por cento de redução da viabilidade de células HeLa em 24 h. Nossos resultados sugerem que eupafolina e hispidulina têm efeitos citotóxicos que podem ser explicados em parte pelas alterações promovidas por estas flavonas sobre propriedades de membranas biológicas e sobre a bioenergética mitocondrial.