Convallatoxin induces HaCaT cell necroptosis and ameliorates skin lesions in psoriasis-like mouse models.

Psoriasis is considered an immune-mediated inflammatory skin disorder that affects the quality of life of nearly four percent of the world population. Considering the side effects of existing therapeutic drugs and the urgent need for new drug development, we screened more than 250 traditional Chinese medicine compounds to identify drugs that significantly reduced the viability of human HaCaT keratinocytes, a psoriasis-related model cell line. Convallatoxin (CNT) was found to be a highly effective inhibitor of HaCaT cell viability. Subsequent mechanistic studies revealed that CNT induced HaCaT cell death by necroptosis rather than by apoptosis. CNT destroyed the membrane integrity of HaCaT cells, as detected by nuclear propidium iodide (PI) staining and lactate dehydrogenase (LDH) release. Additionally, the intercellular levels of adenosine triphosphate (ATP) were lower in HaCaT cells treated with CNT than in control HaCaT cells, and typical necroptosis-associated characteristics were observed by electron microscopy in cells treated with CNT. Furthermore, compared with control HaCaT cells, CNT-treated HaCaT cells produced more reactive oxygen species (ROS), but this effect was inhibited by the antioxidants N-acetyl-cysteine (NAC), diphenyleneiodonium chloride (DPI), and apocynin and the necroptosis inhibitor Nec-1. In addition, antioxidant treatment attenuated necroptotic cell death, suggesting that CNT-induced HaCaT necroptosis is mediated by oxidative stress. More importantly, CNT ameliorated skin lesions and inflammation in imiquimod (IMQ)- and 12-O-tetradecanoyl-phorbol-13-acetate (TPA)-induced psoriasis-like mouse models. In conclusion, our results demonstrate that CNT is cytotoxic against HaCaT cells in vitro and exerts antipsoriatic activities in two mouse models of psoriasis in vivo, making CNT a potential promising candidate drug for future research.

Convallatoxin protects against dextran sulfate sodium-induced experimental colitis in mice by inhibiting NF-κB signaling through activation of PPARγ.

Convallatoxin (CNT) is a cardiac glycoside isolated from Adonis amurensis Regel et Radde and has both anti-inflammatory and anti-proliferative properties. In the present study, the anti-inflammatory mechanisms of action of CNT was investigated in vitro and in vivo. Stimulation of mouse macrophages with lipopolysaccharide induced secretion of proinflammatory cytokines via suppression of peroxisome proliferator-activated receptor gamma (PPARγ) and activation of nuclear factor-κB (NF-κB), two transcription factors implicated in many inflammatory diseases. Notably, the effects of lipopolysaccharide were reversed by concomitant treatment of macrophages with CNT. Knockdown of PPARγ by siRNA inhibited the effect of convallatoxin on NF-κB activation. Because these transcription factors play a role in the development of ulcerative colitis in humans, the mice with experimental colitis induced by dextran sodium sulfate (DSS) was employed. Indeed, concomitant treatment with CNT ameliorated DSS-induced colitis symptoms, tissue damage, inflammatory cell infiltration, and proinflammatory cytokine production in the colon, and also reversed the activation of NF-κB and suppression of PPARγ. Collectively, these data indicate that CNT ameliorates colitic inflammation via activation of PPARγ and suppression of NF-κB, and suggest that CNT may be a promising treatment for inflammatory bowel disease (IBD).

Convallatoxin, a dual inducer of autophagy and apoptosis, inhibits angiogenesis in vitro and in vivo.

Autophagy and apoptosis are important processes that control cellular homeostasis and have been highlighted as promising targets for novel cancer therapies. Here, we identified convallatoxin (CNT), isolated from Antiaris toxicaria, as a dual inducer of autophagy and apoptosis. CNT exerts cytotoxic effects on a number of cancer and normal cell lines and induces apoptosis by increasing caspase-3 and poly ADP ribose polymerase (PARP) cleavage. Moreover, dose- and time-dependent autophagic activity was detected in CNT-treated cells, and mammalian target of rapamycin (mTOR)/p70S6K signal pathway inhibition was observed. Notably, CNT inhibits human umbilical vein endothelial cell (HUVEC) growth and exerts anti-angiogenic activity in vitro and in vivo. Collectively, these results demonstrate that the naturally occurring compound, CNT, is a novel anti-angiogenic compound via dual inducing of autophagy and apoptosis.

Protein-protein interaction network analysis in chronic obstructive pulmonary disease.

BACKGROUND: The aim of this study was to investigate the gene expression profile of chronic obstructive pulmonary disease (COPD) patients and non-COPD patients. METHODS: Microarray raw data (GSE29133) was downloaded from Gene Expression Omnibus, including three COPD samples and three normal controls. Gene expression profiling was performed using Affymetrix human genome u133 plus 2.0 GeneChip. Differentially expressed genes were identified by Student's t test and genes with p < 0.05 were considered significantly changed. Up- and downregulated genes were submitted to the molecular signatures database (MSigDB) to search for a possible association with other previously published gene expression signatures. Furthermore, we constructed a COPD protein-protein interaction (PPI) network and used the connectivity map (cMap) to query for potential drugs for COPD. RESULTS: A total of 680 upregulated genes and 530 downregulated genes in COPD were identified. The MSigDB investigation found that upregulated genes were highly similar to gene signatures that respond to interferon and downregulated genes were similar to erythroid progenitor cells from fetal livers of E13.5 embryos with KLF1 knocked out. A PPI network consisting of 814 gene/proteins and 2,613 interactions was identified by Search Tool for the Retrieval of Interacting Genes. The cMap predicted helveticoside, disulfiram, and lanatoside C as the top three possible drugs that could perhaps treat COPD. CONCLUSION: Comprehensive analysis of the gene expression profile for COPD versus control reveals helveticoside, disulfiram, and lanatoside C as potential molecular targets in COPD. This evidence provides a new breakthrough in the medical treatment of patients with COPD.

[The effect of kavinton and korglycon on the central and cerebral hemodynamics of patients with an ischemic stroke in stenoses and occlusions of the major cerebral arteries]. / Vplyv kavintonu i korhlikonu na tsentral'nu ta tserebral'nu hemodynamiku u khvorykh na ishemichnyi insul't pry stenozakh i okliuziiakh mahistral'nykh arterii holovy.

Of the 40 patients, stenoses of major brain arteries were identified in 34, occlusions in 6, affection of two arteries in eleven patients, that of three arteries in 3 patients. The following disturbances in central hemodynamics were detected: arterial pressure getting higher, this referring also to the peripheral vascular resistance, hematocrit value, with the bloodflow in the arm-ear portion slowing down, hemodynamics being predominantly of hyperkinetic type. Treatment with cavinton, cavinton plus corglucone resulted in improvement of bloodflow in the affected major brain arteries at the expense of a compensating redistribution of bloodflow from intact collectors to the stenosed arteries basin. Changes in central hemodynamics were manifested by increase in the number of cases presenting with eukinetic-type circulation at the expense of decrease in those of hyperkinetic type. Under hypokinetic type hemodynamics, it is advisable that cavinton be prescribed in combination with cardiac glycosides.

[The effect of the glycoside strophanthin K on the ultrastructure of the cardiomyocytes in the pro-oxidant zone of the myocardium in rats with experimental heart failure]. / Vliianie glikozida strofantina K na ul'trastrukturu kardiomiotsitov prooksidantnoi zony miokarda u krys s éksperimental'noi serdechnoi nedostatochnost'iu.

The effect of the cardiotonic strophanthin K on the ultrastructure of cardiomyocytes in the "pro-oxidant zone" in hypercatecholemia was studied in Wistar rats with experimental cardiac insufficiency (ECI). In ECI two differently directed processes, compensatory and degenerative, develop in the "pro-oxidant zone". After administration of strophanthin K the degenerative processes in the "pro-oxidant zone" increased: the number of degenerated cardiomyocytes was 41.9 +/- 1.4% in ECI and 85.35 +/- 3.4% after strophanthin administration (p > 0.99).

[The pentoxifylline and corglycon treatment of patients with circulatory encephalopathy against a background of atherosclerotic lesions of the major cerebral arteries]. / Likuvannia pentoksyfilinom i korhlikonom khvorykh na dystsyrkuliatornu entsefalopatiiu na foni aterosklerotychnoho urazhennia mahistral'nykh arterii holovy.

As many as 34 patients having stage II-III atherosclerotic dyscirculatory encephalopathy with coexisting ischemic heart disease in the presence of stenosing and occlusive lesions of major brain arteries were evaluated for effects pentoxiphyllin and corglycon on the clinical course of this medical condition as well as systemic and cerebral haemodynamics. In this series, 82.7 percent of patients derived benefit from a single intravenous infusion of pentoxiphyllin and corglycon and course treatment with these drug preparation that was evidenced by an improvement in the parameters of systemic and cerebral hemodynamics and regression of neurologic symptomatology. The results of the studies made permit recommending pentoxiphyllin in combination with corglycon for treatment of those DE patients with eukinetic and hypokinetic types systemic haemodynamics, whereas pentoxiphyllin alone--for those with hyperkinetic type of haemodynamics, taking into consideration its cardiodepressive effects.

Long-term use of K-strophanthin in advanced congestive heart failure due to dilated cardiomyopathy: a double-blind crossover evaluation versus digoxin.

K-strophanthin or digoxin were added to diuretics (all cases) and vasodilators (most cases) for treating advanced congestive heart failure in 22 patients with dilated cardiomyopathy and sinus rhythm. K-strophanthin (0.125 mg intravenously) or digoxin (0.25 mg orally) were administered daily in two 3-month periods, during which vasodilators and diuretics were kept constant and patients received one of the two digitalis preparations in a double-blind fashion, crossing over to the alternative preparation in the next period. Blindness was assured throughout the trial with a daily intravenous injection of 10 ml normal saline solution either containing K-strophanthin or not, and with daily oral administration of either placebo or active digoxin. At the end of the run-in period, 15 days after starting active preparations, and thereafter every month for the next 6 months, we evaluated left ventricular pump function at rest and patients' functional performance by a cardiopulmonary exercise test. At Day 15, cardiac index and ejection fraction at rest, compared with run-in, were significantly raised with both glycosides; during exercise while on K-strophanthin, peak oxygen consumption was augmented by 1.4 ml/min/kg (p < 0.01) and oxygen consumption at anaerobic threshold by 2.2 ml/min/kg (p < 0.01); corresponding variations on digoxin (-0.1 and +0.3, respectively) were not significant versus run-in. These patterns were duplicated at repeated tests during follow-up.(ABSTRACT TRUNCATED AT 250 WORDS)

[Diabetic myocardiopathy].

Three cases of diabetic myocardiopathy having history of diabetes, angina and left ventricular dysfunction of various degrees and confirmed by coronary angiography and endomyocardial biopsy were reported. Electrocardiography showed significant ST-T changes simulating coronary insufficiency but without definite localization. As to the treatment, nitrate preparations, inotropic agents such as strophanthin K, digoxin etc. were used to relieve the symptoms; insulin was also administered to control the blood glucose level. Diltiazem, a calcium blocker, is also of help in alleviating the symptoms. It is shown in the present study and in the literatures as well that diabetic myocardiopathy is a disease showing intramural microvascular endothelial proliferation and swelling as well as subendothelial accumulation of acid glycogen deposition cells. The transportation of intracellular calcium ions and the cellular metabolism are thus affected, so there are extensive ischemia, focal necrosis and fibrosis in the myocardium with resulting cardiac dysfunction. The authors are, therefore, of the opinion that diabetic myocardiopathy is a specific and separate clinical entity.

[The influence of kordaron on the pharmacodynamic effects of strophanthin in experiments on isolated myocardial preparations and in the modelling of heart failure]. / Vliianie kordarona na farmakodinamicheskie éffekty strofantina v éksperimentakh na izolirovannykh preparatakh miokarda i v usloviiakh modelirovaniia serdechnoi nedostatochnosti.

The experiments with isolated rat atria isometrically contracting and those with simulated rat heart failure were performed to study the effects of the alpha-, beta-, and X-blocker cordarone on the pharmacodynamic effects of strophanthin. Cordarone was demonstrated to greatly decrease the toxicity of the cardiotonic in circulatory decompensation, without causing negative effects of cardiac inotropic function. Cordarone in combination with strophanthin slightly diminished the magnitude of the negative chronotropic effect of the cardiac glycoside and slowed down the rate of its cardiotonic effect.

[Effect of isoptin on toxic and cardiotonic effect of strophanthin K]. / Vliianie izoptina na toksicheskii i kardiotonicheskii éffekty strofantina K.

Two models of heart failure were used in rat experiments to study the effect of isoptin on tolerance to the toxic effect of k-strophanthin. Calcium antagonist-induced changes in the cardiotonic and chronotropic effects of the cardiac glycoside and myocardial oxygen consumption were examined in isolated frog atria with a movable-electrode electron tube and polarography. Isoptin was shown to substantially enhance tolerance to the toxic effect of k-strophanthin and slightly decrease positive inotropic effects of the cardiotonic, by slowing down its development rate. The agent potentiated the negative chronotropic action and lowered myocardial oxygen consumption.

[Effects of xin-fu-ning IV on experimental congestive heart failure in rabbits].

By use of pentobarbital sodium to produce experimental congestive heart failure (CHF) in 40 rabbits, they were treated with xin-fu-ning IV (XFN IV), the plasma atrial natriuretic factor (ANF) levels and atrial tissue ANF levels of rabbits were measured. The results showed that XFN IV had elevated ANF levels of the rabbits with CHF. It showed that XFN IV could improve endocrine function of heart, and it also showed good curative effect on CHF.

[Sodium nitroprusside in treating acute high-altitude pulmonary edema].

Sodium nitroprusside (NP) alone Was given to the 39 cases with acute high-altitude pulmonary edema (AHPE). 33 of them (84.6%) fully recovered and 6 (15.38%) obviously improved in 72 hours. The total efficacy was 100%. There was obvious improvement in the cardiac pumping and contractile function as well as total peripheral resistance. The mean pulmonary artery pressure (MPAP) decreased from 52.22 +/- 7.51mmHg to 25.13 +/- 10.36mmHg in 3 to 5 hours after the first dose. Strophanthin K was given to another 11 cases with AHPE, But none fully recovered or obviously improved. During the 72 hours of treatment, 4 improved slightly, 4 showed no therapeutic effect and 3 became worse. The heart rate was reduced obviously and cardiac contractile function strengthened while the cardiac pumping function and total peripheral resistance had no change. Vasodilator therapy is a new method for AHPE treatment and this study shows that the NP is an ideal drug in treating AHPE and that dilation of the small pulmonary arteries by NP may explain its efficacy.

[Effect of strophanthin and digoxin on the activity of succinate and lactate dehydrogenases and membrane Na+, K+-ATPase in the heart of rats with experimental myocarditis]. / Vliianie strofantina i digoksina na aktivnost' suktsinat-laktatdegidrogenaz i membrannoi Na+, K+-ATFazy v serdtse krys s éksperimental'nym miokarditom.

In rats with adrenaline-induced myocarditis conditionally therapeutic doses of strophanthin (2.7 mg/kg) and digoxin (0.89 mg/kg) were chosen according to performance of the test of swimming until the complete fatigue. The influence of drugs in these doses on enzymatic activity was evaluated by histochemical methods in heart of control and myocarditis rats. It was found out that both of cardiac glycosides decreased lactate dehydrogenase and membrane Na+, K+-ATPase activity and increased succinate dehydrogenase activity in rats with experimental myocarditis.

[Myocardial contractile function in acute heart failure during exposure to strophanthin and its combination with alpha-tocopherol]. / Sokratitel'naia funktsiia miokarda pri ostroi serdechnoi nedostatochnosti v usloviiakh deistviia strofantina i ego sochetanii s al'fa-tokoferolom.

In experiments on rabbits it was established that strophanthin (0.05 mg/kg) administered to animals with cardiac insufficiency due to a severe overloading of the heart function by occlusion of the diaphragmatic portion of the aorta by 4/5 of its diameter improved the contractile activity and relaxing properties of the left ventricle. This effect is however transient (up to 15-30 minutes) with the subsequent reduction of its characteristic indices, the development of symptoms of intoxication and not infrequently death of animals. alpha-tocopherol administered before the pathology reproduction (10 mg/kg, 3 days) increases the period of myocardial compensation and when given in combination with strophanthin significantly potentiates the cardiac effect of glycoside manifesting by an increase of the strength and rate of contraction and intensity of diastolic relaxation of the myocardium, prolongs the animal's life.

[Correction of hemodynamics in patients with acute pneumonia]. / Korrektsiia gemodinamiki u bol'nykh ostroi pnevmoniei.

Patients with acute pneumonia demonstrate disturbance of myocardial contractility manifesting itself in an increase of end diastolic and systolic volumes, a decrease in the ejection fraction and circular myocardial contractility rate. In a raised level of mean arterial pressure i.v. administration of strophanthin in acute pneumonia can produce a negative effect in the form of a decrease in the stroke volume and cardiac output. Myofedrin promotes an increase in cardiac ejection and can be used for therapy of patients with acute pneumonia to correct hemodynamics.