Objective: To study the effects of dihydromyricetin (DMY) on the proliferation, apoptosis and epithelial mesenchymal transition (EMT) of esophageal squamous cell carcinoma (ESCC) cell KYSE150 and KYSE410. Methods: KYSE150 and KYSE410 cells were treated with different concentrations of DMY (0, 25, 50, 100, 150, 200 μmol/L) for 24 hours. The median inhibition concentration (IC₅₀) values of KYSE150 and KYSE410 were detected by cell counting kit-8 (CCK-8) method. Then 0.5‰ dimethyl sulfoxide (DMSO) was used as control group, dihydromyricetin (DMY), dihydromyricetin and transforming growth factor-β1 (DMY+ TGF-β1), transforming growth factor-β1 (TGF-β1) were used as experimental group. Cell proliferation and apoptosis rates were measured by clonal formation and flow cytometry. Transwell invasion and wound healing assay were used to detect cell invasion and migration. The protein expression levels of Caspase-3, Caspase-9, Bcl-2, Bax, Smad2/3, phosphorylation-Smad2/3 (p-Smad2/3) and Vimentin were detected by western blot. Results: The IC₅₀ values of DMY on KYSE410 and KYSE150 cells were 100.51 and 101.27 μmol/L. The clone formation numbers of KYSE150 and KYSE410 in DMY group [(0.53±0.03) and (0.31±0.03)] were lower than those in DMSO group [(1.00±0.10) and (1.00±0.05), P<0.05]. The apoptosis rates of KYSE150 and KYSE410 cells in DMY group [(1.84±0.22)% and (2.80±0.07)%] were higher than those in DMSO group [(1.00±0.18)% and (1.00±0.07)%, P<0.05]. The invasion numbers of KYSE150 and KYSE410 cells in DMY group [(0.42±0.03) and (0.29±0.05)] were lower than those in DMSO group [(1.00±0.08) and (1.00±0.05), P<0.05]. The migration rates of KYSE150 and KYSE410 cells in DMY group [(0.65±0.14)% and (0.40±0.17)%] were lower than those in DMSO group [(1.00±0.10)% and (1.00±0.08)%, P<0.05]. The clone formation numbers of KYSE150 and KYSE410 in TGF-β1 group [(1.01±0.08) and (0.99±0.25)] were higher than those in DMY+ TGF-β1 group [(0.73±0.10) and (0.58±0.05), P<0.05]. The apoptosis rates of KYSE150 and KYSE410 cells in TGF-β1 group [(0.81±0.14)% and (1.18±0.10)%] were lower than those in DMY+ TGF-β1 group [(1.38±0.22)% and (1.85±0.04)%, P<0.05]. The invasion numbers of KYSE150 and KYSE410 cells in TGF-β1 group [(1.19±0.11) and (1.39±0.11)] were higher than those in DMY+ TGF-β1 group [(0.93±0.09) and (0.93±0.05), P<0.05]. The migration rates of KYSE150 and KYSE410 cells in TGF-β1 group [(1.87±0.19)% and (1.32±0.04)%] were higher than those in DMY+ TGF-β1 group [(0.86±0.16)% and (0.77±0.12)%, P<0.05]. The protein expression levels of Bax, Caspase-3 and Caspase-9 in KYSE150 and KYSE410 cells in DMY group were higher than those in DMSO group, while the protein expression level of Bcl-2 was lower than that in DMSO group (P<0.05). The protein expression levels of p-Smad2/3, Smad2/3 and Vimentin in KYSE150 and KYSE410 cells in DMY group were lower than those in DMSO group (P<0.05). The protein expression levels of Bax, Caspase-3 and Caspase-9 in KYSE150 and KYSE410 cells in TGF-β1 group were lower than those in DMY+ TGF-β1 group, and the protein expression level of Bcl-2 was higher than that in DMY+ TGF-β1 group (P<0.05). The protein expression levels of Bax, Caspase-3 and Caspase-9 in KYSE150 and KYSE410 cells in DMY+ TGF-β1 group were lower than those in DMY group, and the protein expression level of Bcl-2 was higher than that in DMY group (P<0.05). The protein expression levels of p-Smad2/3, Smad2/3 and Vimentin in KYSE150 and KYSE410 cells in TGF-β1 group were higher than those in DMY+ TGF-β1 group (P<0.05). Conclusion: DMY can inhibit the proliferation and EMT of ESCC mediated by TGF-β1 and promote cell apoptosis.
Apoptosis ; Caspase 3/metabolism* ; Caspase 9/metabolism* ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Dimethyl Sulfoxide/pharmacology* ; Epithelial-Mesenchymal Transition ; Esophageal Neoplasms/metabolism* ; Esophageal Squamous Cell Carcinoma ; Flavonols ; Humans ; Signal Transduction ; Transforming Growth Factor beta1/pharmacology* ; Vimentin/metabolism* ; bcl-2-Associated X Protein/pharmacology*

OBJECTIVE: To explore the effect of dihydromyricetin on the expression of miR-98-5p and its mechanism in the development of Herceptin resistance in SKBR3 cells. METHODS: The expression of IGF2 and miR-98-5p and their interaction relationship were analyzed by bioinformatics analysis through TargetScan online databases. SKBR3 cells and drug-resistant SKBR3-R cells were cultured in cell experiments. Xenograft tumor mice were constructed by SKBR3 and SKBR3-R cells. Proteins were detected by western blotting and immunohistochemistry. Transfected cells were constructed by shRNA lentivirus vectors. RT-QPCR was used to detect RNA. Cell proliferation was detected by MTS method. Cell jnvasion was detected by Transwell assay. Luciferase reporting assays were used to verify RNA interactions. IGF-1R/HER2 heterodimer was determined by immunocoprecipitation. RESULTS: The expression of IGF2, p-IGF1R, p-Akt and p-S6K in SKBR3-R cells were significantly higher than those in SKBR3 cells, while the expression of PTEN protein was lower in SKBR3-R cells (P < 0.05). IGF1R/HER2 heterodimer in SKBR3-R cells was significantly increased (P < 0.01).The expression of IGF2 and invasion ability were significantly reduced while transfected with miR-98-5p in SKBR3-R cells (P < 0.05), but the IGF2 mRNA were no difference in both cells (P > 0.05). The expression of miR-98-5p was up-regulated and IGF2 was decreased in drug-resistant xenograft tumor mice after feeding with dihydromyricetin, and the tumor became more sensitivity to Herceptin (P < 0.05). CONCLUSION Dihydromyricetin could induce the expression of miR-98-5p, which binds to IGF2 mRNA to reduce IGF2 expression, inhibit the IGF-1R/HER2 formation, thereby reversing cell resistance to Herceptin in SKBR3-R cells.
Animals ; Cell Line, Tumor ; Flavonols/pharmacology* ; Humans ; Mice ; MicroRNAs/metabolism* ; Receptor, IGF Type 1 ; Trastuzumab

Ten compounds, including five steroidal saponins and five flavonol glycosides, were isolated from the whole plant of Solanum coagulans by means of column chromatographies over silica gel, ODS, Sephadex LH-20, and preparative HPLC. Based on analysis of MS and NMR spectroscopic data, their structures were established as anguiviosides XV (1), smilaxchinoside A (2), methylprotodioscin (3), protodioscin (4), solamargine (5), 3', 4', 5-trihydroxy-7-methoxy-6-C-β-D-glucopyranoside (6), brainoside B (7), camsibriside A (8), kampferol 3-O-(2"-β-D-galactopyranosyl)-β-D-glucopyranoside (9), and quercetin-3-O-(2"-β-D-galactopyranosyl)-β-D-glucopyranoside (10). All the compounds were first isolated from this plant. In the in vitro assays, compounds 4 and 5 showed cytotoxic activity against SMCC-7721 and NCI-H460.
Cell Line, Tumor ; Flavonols ; chemistry ; isolation & purification ; pharmacology ; Humans ; Saponins ; chemistry ; isolation & purification ; pharmacology ; Solanum ; chemistry

OBJECTIVEAstilbin in 28 Smilax glabra (red and white cross-section) from different sources was determined by HPLC. Pharmacodynamics and component of S. glabra was investigated through inflammation experiment (penetration type).

METHODThe analysis was performed on a Hypersil ODS2 column (4.6 mm x 250 mm, 5 microm) with the mobile phase of acetonitrile and 0. 1% acetic acid aqueous (21: 79) at a flow rate of 1.0 mL x min(-1). The detection wavelength was 291 nm, and the column temperature was 25 degrees C. Anti-inflammatory effect was compared from two type cross-section of Smilax glabra in capillary permeability experiment.

RESULTLinear correlation was good in the range of 0.003 379-4.004 microg, and the average recoveries were 100.1%, 101.9%, 99.3%, respectively. The content of astilbin in white and red transverse section were 0.19%-2.46% and 2.10%-5.92%, respectively. Anti-inflammatory efficiency of sectioned red and white were were 21% and 32%, respectively.

CONCLUSIONAstilbin content and anti-inflammatory effect is significantly different between red and white transverse section of S. glabra, the content of astilbin is not positively related with anti-inflammatory effect.

Animals ; Anti-Inflammatory Agents ; chemistry ; pharmacology ; China ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Flavonols ; chemistry ; pharmacology ; Male ; Mice ; Permeability ; Smilax ; chemistry

OBJECTIVETo observe the effect of serum containing Tengcha total flavonoid and dihydromyricetin on proliferation and apoptosis of HepG2 cells.

METHODSerum containing respectively Tengcha total flavonid, dihydromyricetins and CTX and control serum were prepared by serological pharmacology method. MTT assay was used to observe the proliferation inhibition rate of HepG2 cells after incubated with different kinds of serum. Inverted microscope was utilized to observe the morphological changes after HepG2 cells were treated with different serum. AnnexinV/7AAD double label method was used to detect earlier period apoptosis cells.

RESULTBoth serum containing 20% Tengcha total flavonid and serum containing 20% dihydromyricetin could restrain the HepG2 cells proliferation at different levels and the morpholological changes of apoptosis were observed. AnnexinV/7AAD double label method showed that the earlier period apoptosis cells rates were increased by serum containing 20% Tengcha total flavonoid, but serum containing 20% dihydromyricetin did not show influence on the earlier period apoptosis cells.

CONCLUSIONTengcha total flavonoid can restrain the HepG2 cells proliferation and induce earlier period apoptosis cells.

Ampelopsis ; chemistry ; Animals ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Flavonoids ; blood ; pharmacology ; Flavonols ; blood ; pharmacology ; Hep G2 Cells ; drug effects ; Humans ; Male ; Rats ; Rats, Wistar

OBJECTIVESTo investigate the effects of astilbin on the expressions of TNF alpha and IL-10 during liver warm ischemia-reperfusion injury.

METHODSC57BL/ 6 mice were randomly divided into 4 groups (n = 8): sham-operated group (Sham), model control group(I/R), low dosage of astilbin treatment group (10 mg/kg) and high dosage of astilbin (40 mg/kg) treatment group. The treatment group mice were intraperitoneally injected with 10 or 40 mg/kg astilbin 24 hours and one hour before Ischemia, the hepatic ischemia-reperfusion model were thus established. After jn90 of min ischemia and 6 h reperfusion of the partial hepatic lobe, the expressions of TNF alpha and IL-10 in liver tissues collected from the experimental groups were detected by Western blot and semiquantitative RT-PCR.

RESULTSThe expression of TNF alpha protein in liver tissues gradually decreased in treatment groups (low and high dosages of astilbin treatment groups) as compared to the I/R model control group. Similar results were observed in the mRNA expressions of these genes as determined by semiquantitative RT-PCR (P less than 0.05 for low dosage group; P less than 0.01 for high dosage group). Compared with the I/R model control group, the expression of IL-10 was increased in both treatment groups (low dosage group P less than 0.05; large dosage group P less than 0.01).

CONCLUSIONTreatment with astilbin decreases TNF alpha expression but induces IL-10 expression in liver during warm ischemia-reperfusion injury.

Animals ; Flavonols ; pharmacology ; Interleukin-10 ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Reperfusion Injury ; etiology ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Warm Ischemia

BACKGROUNDMyocyte apoptosis is considered to be the major causative factor of left ventricular (LV) remodeling following myocardial infarction (MI). We previously reported that 3', 4'-dihydroxyflavonol (DiOHF), was able to suppress oxidative stress and preserve the expression of endothelial nitric oxide synthase during myocardial reperfusion injury, which may benefit the reduction of myocyte apoptosis. We therefore aimed to evaluate the potential actions of DiOHF against myocyte apoptosis and post-infarction LV remodeling in this study.

METHODSFollowing experimental MI, surgical instrumented goats were randomly assigned into vehicle and DiOHF (2 mg/kg; i.v., daily) groups to receive 4 weeks of reperfusion with corresponding treatments. LV pressure recordings and echocardiogram were performed at baseline, 2 and 4 weeks of reperfusion. Myocardial tissues were collected in the end to determine infarct size and apoptosis related assays.

RESULTSLV end-diastolic volume and diameter were significantly increased 4 weeks after MI in the vehicle group, accompanied by reduced posterior wall thickness, septal thickness and LV mass, whereas those changes were markedly prevented by DiOHF treatment. Similarly, significantly reduced infarct size was found in DiOHF group as compared to vehicle group, and DiOHF dramatically inhibited the increase in LV end-diastolic pressure and the reductions in ejection fraction, fraction shortening and dP/dt(max). Moreover, DiOHF treatment significantly reduced the extent of myocyte apoptosis detected by TUNEL assay, enhanced the protein expression of caspase-3, Fas, Bax and cytochrome c in the non-infarcted myocardium in comparison to vehicle.

CONCLUSIONSDaily DiOHF treatment during the reperfusion period after MI in the ovine hearts markedly reduced the magnitude of post-infarction LV remodeling through the inhibition of myocyte apoptosis in the remote non-infarcted myocardium.

Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cytochromes c ; metabolism ; Echocardiography ; Female ; Flavonols ; pharmacology ; Goats ; In Situ Nick-End Labeling ; Male ; Myocardial Infarction ; metabolism ; physiopathology ; Myocytes, Cardiac ; cytology ; drug effects ; Random Allocation ; Ventricular Remodeling ; drug effects ; bcl-2-Associated X Protein ; metabolism ; fas Receptor ; metabolism

Ten compounds were isolated from the leaves of Rhizophora stylosa, one kind of mangrove plants distributed in the tropical and subtropical areas of the world. Their structures were identified as taraxerone (1), taraxerol (2), beta-sitosterol (3), careaborin (4), cis-careaborin (5), beta-daucosterol (6), isovanillic acid (7), protocatechuic acid (8), astilbin (9) and rutin (10), among which compound 9 and 10 were reported in this plant for the first time. Of these compounds, Compound 2 has been confirmed to have the abilities to inhibit the growth of Hela and BGC-823 with IC50 of 73.4 micromol x L(-1) and 73.3 micromol x L(-1), respectively. Compound 5 could inhibit the growth of BGC-823 and MCF-7 with IC50 of 45.9 micromol x L(-1) and 116.0 micromol x L(-1), respectively. Compound 9 and 10 were firstly reported to stimulate the proliferation of mice splenic lymphocytes markedly in a dose-dependent manner.
Animals ; Antineoplastic Agents, Phytogenic ; chemistry ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Flavonols ; chemistry ; isolation & purification ; pharmacology ; Humans ; Inhibitory Concentration 50 ; Lymphocytes ; cytology ; Mice ; Molecular Structure ; Oleanolic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; pharmacology ; Plant Leaves ; chemistry ; Rhizophoraceae ; chemistry ; Rutin ; chemistry ; isolation & purification ; pharmacology ; Spleen ; cytology ; Triterpenes ; chemistry ; isolation & purification ; pharmacology

This study was focused on the circadian rhythms of DNA synthesis and the expression of c-myc gene in untreated and treated Eca-109 cells in human oesophageal cancer with isorhmnetin. The circadian rhythms of 3H-TdR incorporation and expression of c-myc gene in untreated and treated Eca-109 cells were measured by 3H-thymidine uptake assay and flow cytometry. The data collected were analyzed by ANOVA and Cosinor method. DNA synthesis and expression of c-myc gene in untreated group varied according to circadian time with statistical significance, the distribution curves of both DNA synthesis and the expression level of c-myc were fit for cosinor changes. The circadian rhythms of DNA synthesis and circadian parameters of c-myc expression in treated Eca-109 cells changed. The circadian parameters of DNA synthesis and expression level of c-myc varied after treatment by isorhmnetin. The effects of isorhmnetin on cell proliferation and c-myc expression reached the highest level from 20: 00 to 0: 00. The results provide a guidance for instituting the chemotherapy and chronotherapy of human tumors, when isorhmnetin is for use as anti-cancer agent.
Circadian Rhythm ; drug effects ; DNA ; biosynthesis ; Esophageal Neoplasms ; genetics ; metabolism ; pathology ; Flavonols ; pharmacology ; Humans ; Proto-Oncogene Proteins c-myc ; biosynthesis ; genetics ; Quercetin ; analogs & derivatives ; Tumor Cells, Cultured

OBJECTIVETo explore the effects of total flavonoids of Hippophae rhamnoides L. (TFH), quercetin (Que) and isorhamnetin (Isor) on the intracellular free calcium ([Ca(2+)](i)) in vascular smooth muscle cells (VSMC) of spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY).

METHODSFluo 3-acetoxymethylester (Fluo-3/AM) was used to observe the effects of TFH (100 mg/L) and its essential monomers, namely Que (10(-4) mol/L) and Isor (10(-4) mol/L) on changes of [Ca(2+)](i) in cultured SHR and WKY VSMC (abbr. to Ca-SHR & Ca-WKY) following exposure to high K(+), norepinephrine (NE) and angiotensin II (Ang II), and to compare with the effects of verapamil (Ver).

RESULTS(1) TFH, Que and Isor had inhibitory effects on resting Ca-SHR (P < 0.05), but had no significant effects on Ca-WKY (P > 0.05). (2) High K(+) could increase Ca-SHR more significantly than Ca-WKY (P < 0.05); TFH, Que and Isor could inhibit the elevation of [Ca(2+)](i) induced by high K(+)-depolarization, with the effects similar to that of Ver, and the effect on Ca-SHR was more significant than that on Ca-WKY (P < 0.05). (3) NE and Ang II could increase Ca-SHR more significantly than Ca-WKY (P < 0.05), TFH, Que and Isor had remarkably inhibitory effect on the elevation of Ca-SHR and Ca-WKY induced by NE or Ang II. (4) In the absence of extracellular Ca(2+), TFH, Que and Isor also had certain inhibitory effect on Ca-SHR and Ca-WKY induced by NE, and the effect on the former was more significant than that on the latter (P < 0.05).

CONCLUSIONTFH, Que and Isor might decrease the levels of [Ca(2+)](i) in VSMCs by blocking both voltage-dependent calcium channels (VDC) and receptor-operated calcium channels (ROC) in physiological or pathological state, which may be one of the important mechanisms of their hypotensive and protective effects on target organs in patients with hypertension.

Angiotensin II ; pharmacology ; Animals ; Calcium ; analysis ; Cells, Cultured ; Flavonoids ; pharmacology ; Flavonols ; pharmacology ; Hippophae ; Hypertension ; metabolism ; Muscle, Smooth, Vascular ; chemistry ; cytology ; Norepinephrine ; pharmacology ; Quercetin ; pharmacology ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Verapamil ; pharmacology