Hard antler extract inhibits invasion and epithelial-mesenchymal transition of triple-negative and Her-2+ breast cancer cells by attenuating nuclear factor-κB signaling.

ETHNOPHARMACOLOGICAL RELEVANCE: Hard antler extract (HAE) is a traditional Chinese medicine and has potent antitumor, antioxidative, anti-inflammatory, and immunomodulatory activities. Previous studies have demonstrated that HAE can inhibit human prostate cancer metastasis and murine breast cancer proliferation. However, the effect of HAE on human breast cancer cells has not been clarified. AIM OF THE STUDY: To investigate the effects and underlying mechanism of HAE on self-renewal of stem-like cells and spontaneous and transforming growth factor (TGF)-ß1-enhanced wound healing, invasion and epithelial-mesenchymal transition (EMT) in breast cancer cells. METHODS: HAE was prepared from sika deer by sequential enzymatic digestions and the active compounds were determined by HPLC. The effects of HAE on the viability, mammosphere formation, wound healing and invasion of MDA-MB-231 and SK-BR3 cells were determined. The impact of HAE treatment on spontaneous and TGF-ß1-promoted EMT and the nuclear factor (NF)-κB signaling in breast cancer cells was examined by quantitative RT-PCR and western blotting. RESULTS: Treatment with HAE at varying concentrations did not change the viability of breast cancer cells. However, HAE at 0.25 or 0.5 mg/mL significantly reduced the number and size of formed mammospheres, and inhibited spontaneous and TGF-ß1-enhanced wound healing, invasion and EMT in MDA-MB-231 and SK-BR3 cells in a dose-dependent manner. TGF-ß1 treatment significantly decreased IκBα expression and increased NF-kBp65 phosphorylation in breast cancer cells, indicating that TGF-ß1 enhanced NF-κB signaling. In contrast, HAE treatment attenuated the spontaneous and TGF-ß1-enhanced NF-κB signaling in breast cancer cells. CONCLUSION: Our data indicated that HAE inhibited the self-renewal of stem-like cells and spontaneous and TGF-ß1-enhanced wound healing, invasion and EMT in breast cancer cells by attenuating the NF-κB signaling in vitro.

Icariin induces apoptosis of human lung adenocarcinoma cells by activating the mitochondrial apoptotic pathway.

Lung cancer is the largest cause of morbidity and mortality among tumor diseases. Traditional first-line chemotherapeutic drugs are frequently accompanied by serious side effects when used to treat tumors, thus, novel drugs with reduced toxic effects may improve a patients' quality of life. Icariin, an extract of herba epimedii, has been demonstrated to exhibit multiple antitumor effects with low toxicity. In the present study, cell cycle analysis, apoptosis assays, DAPI staining, CCK8 assays, xenograft tumor models, mitochondrial membrane potential analysis, western blotting and reverse transcription-quantitative PCR were performed to determine the molecular mechanism underlying icariin activity in the human lung adenocarcinoma cell lines, A549 and H1975. The results showed that icariin reduced proliferation of A549 and H1975 cells in a time- and dose-dependent manner in vitro to a greater degree than the control BEAS-2B cells, and this was associated with increased apoptosis, but not with cell cycle progression. In vivo experiments showed that icariin treatment significantly decreased proliferation of H1975 cells in a xenograft mouse model. Mechanistically, icariin activated the mitochondrial pathway by inhibiting the activation of the PI3K-Akt pathway-associated kinase, Akt, resulting in the activation of members of the caspase family of proteins, and thus inducing apoptosis of A549 cells. Taken together, the results revealed that icariin has anti-cancer properties in lung cancer in vitro and in vivo without any noticeable toxic effects on normal lung epithelial cells. Icariin in combination with conventional anti-cancer agents may be an effective therapeutic strategy for treatment of lung carcinoma.

[Research progress on identification and quality control of Cervi Cornu Pantotrichum].

Cervi Cornu Pantotrichum, as a traditional Chinese medicine, has great potential for development. However, the identification and quality control system is not perfect, leading to the market chaos and chronic slow growth in deep processing of Cervi Cornu Pantotrichum. This paper gives an overview of present situation in identification and quality control system of the Cervi Cornu Pantotrichum, and analyzes present problems. Based on these results, the feasibility study scheme in identification and quality control system for Cervi Cornu Pantotrichum would be then put forward, providing ideas to establish its comprehensive evaluation system.

Role of small-conductance calcium-activated potassium channels in atrial electrophysiology and fibrillation in the dog.

BACKGROUND: Recent evidence points to functional Ca²âº-dependent K⁺ (SK) channels in the heart that may govern atrial fibrillation (AF) risk, but the underlying mechanisms are unclear. This study addressed the role of SK channels in atrial repolarization and AF persistence in a canine AF model. METHODS AND RESULTS: Electrophysiological variables were assessed in dogs subjected to atrial remodeling by 7-day atrial tachypacing (AT-P), as well as controls. Ionic currents and single-channel properties were measured in isolated canine atrial cardiomyocytes by patch clamp. NS8593, a putative selective SK blocker, suppressed SK current with an IC50 of ≈5 µmol/L, without affecting Na⁺, Ca²âº, or other K⁺ currents. Whole-cell SK current sensitive to NS8593 was significantly larger in pulmonary vein (PV) versus left atrial (LA) cells, without a difference in SK single-channel open probability (P(o)), whereas AT-P enhanced both whole-cell SK currents and single-channel P(o). SK-current block increased action potential duration in both PV and LA cells after AT-P; but only in PV cells in absence of AT-P. SK2 expression was more abundant at both mRNA and protein levels for PV versus LA in control dogs, in both control and AT-P; AT-P upregulated only SK1 at the protein level. Intravenous administration of NS8593 (5 mg/kg) significantly prolonged atrial refractoriness and reduced AF duration without affecting the Wenckebach cycle length, left ventricular refractoriness, or blood pressure. CONCLUSIONS: SK currents play a role in canine atrial repolarization, are larger in PVs than LA, are enhanced by atrial-tachycardia remodeling, and appear to participate in promoting AF maintenance. These results are relevant to the potential mechanisms underlying the association between SK single-nucleotide polymorphisms and AF and suggest SK blockers as potentially interesting anti-AF drugs.

Effects of Ginkgolide B on action potential and calcium, potassium current in guinea pig ventricular myocytes.

AIM: To investigate the effect of Ginkgolide B (GB) on action potential (AP), delayed rectifier potassium current (IK), and L-type calcium current (I(Ca-L)) in guinea pig ventricular myocytes. METHODS: Single ventricular myocytes were isolated by an enzymatic dissociation method. AP, IK, I(Ca-L) were recorded by whole-cell patch-clamp technique in either current or voltage clamp mode. RESULTS: GB shortened APD in a concentration-dependent manner. GB 0.1, 1, and 10 micromol/L shortened APD50 by 7.9% (n=5, P>0.05), 18.4% (n=5, P<0.01), and 28.9% (n=6, P<0.01), respectively; APD90 by 12.4% (n=5, P>0.05), 17.6% (n=5, P<0.01), 26.4% (n=5, P<0.01), respectively. GB increased IK in a concentration-dependent manner. GB 0.1, 1, and 10 micromol/L increased IK by 20.1% (n=6, P<0.05), 43.1% (n=6, P<0.01), 55.6% (n=6, P<0.05); increased IK tail by 10.7% (n=6, P<0.05), 25.1% (n=6, P<0.05), and 37.7% (n=6, P<0.05), respectively at testing potential of +50 mV and shift the I-V curve of IK upward. But GB had no significant effect on I(Ca-L) at above concentrations. CONCLUSION: GB significantly shortened APD in a concentration-dependent manner which mainly due to increase of IK.

[Effect of liuwei dihuang pill on erythrocyte aldose reductase activity in early diabetic nephropathy patients].

OBJECTIVE: To observe the inhibitory effect of Liuwei Dihuang Pill (LDP) on erythrocyte aldose reductase (EAR) activity in early diabetic nephropathy (DN) patients and to explore the clinical significance of applying LDP in prevention and treatment of DN as an inhibitor of aldose reductase. METHODS: Seventy-two patients diagnosed as early DN with TCM Syndrome of both Qi-yin deficiency were randomly divided into the control group (31 patients) treated by conventional therapy (orally taken Gliquidone or injection of insulin) and the treated group (41 patients) treated by conventional therapy plus LDP for 3 months as one therapeutic course. Changes of symptoms, physical signs, fasting blood glucose (FBG), blood glucose 2 hrs post breakfast (2hPBG), blood total cholesterol (TC), triglyceride (TG), EAR activity, urinary albumin excretion rate (UAER), blood and urinary beta2-microglobulin (beta2-MG) in patients before and after treatment were observed. RESULTS: LDP could improve the symptoms and signs of patients with DN, it could significantly inhibit EAR activity, to make it significantly lower than that in the control group (P < 0.05), and the levels of UAER, beta2-MG in blood and urine in the treated group after treatment were obviously lower than those in the control group (P < 0.05), and LDP showed no apparent effect on blood glucose, lipids and mean arterial pressure (P < 0.05). CONCLUSION: LDP could obviously inhibit the activity of EAR in patients with early DN, improve various indexes of DN, so as to be helpful for its treatment.

[Effect of ginkgolide B on L-type calcium current and cytosolic [Ca2+]i in guinea pig ischemic ventricular myocytes].

With whole-cell variant patch-clamp and laser scanning confocal microscope technique, we examined the effect of ginkgolide B (GB) from ginkgo leaves on L-type calcium current and cytosolic [Ca(2+)](i) in guinea pig ischemic ventricular myocytes. The results showed that under normal conditions, at a test voltage of 0 mV, GB had no significant effect on I(Ca,L); and during ischemia, the peak Ca(2+) current reduced by 37.71%, and the I-V curve of I(Ca,L) was shifted upward. 1 micromol/L GB reversed the change induced by ischemia, a result being significantly different from those of the ishemia group (P<0.05).Under control condition, 0.1,1,10 micromol/L GB decreased intracellular calcium concentration by 10.58%, 17.27% and 16.35% (n=12, 12, 10, P<0.01-0.001), respectively. With perfusion of ischemic solution for 12 min, intracellular calcium concentration increased by 20.15%. After a 12 min-perfusion of ischemic solution containing 1 micromol/L nifedipine or 5 mmol/L NiCl2, intracellular calcium concentration increased by 18.18% (P>0.05 vs ischemia) and 11% (P<0.05 vs ischemia), respectively. After 12 min of perfusion with ischemic solution containing 1 micromol/L GB, intracellular calcium concentration increased by 9.6% (P<0.05 vs ischemia). It is shown that GB could reverse the decrease of I(Ca,L) and partially inhibit calcium overload during ischemia.