Andrographolide mitigates cardiac apoptosis to provide cardio-protection in high-fat-diet-induced obese mice.

Excessive intake of high fat diet (HFD) and associated obese conditions are critical contributors of cardiac diseases. In this study, an active metabolite andrographolide from Andrographis paniculata was found to ameliorate HFD-induced cardiac apoptosis. C57/BL6 mouse were grouped as control (n = 9), obese (n = 8), low dose (25 mg/kg/d) andrographolide treatment (n = 9), and high dose (50 mg/kg/d) andrographolide treatment (n = 9). The control group was provided with standard laboratory chow and the other groups were fed with HFD. Andrographolide was administered through oral gavage for 1 week. Histopathological analysis showed increase in apoptotic nuclei and considerable cardiac-damages in the obese group signifying cardiac remodeling effects. Further, Western blot results showed increase in pro-apoptotic proteins and decrease in the proteins of IGF-1R-survival signaling. However, feeding of andrographolide significantly reduced the cardiac effects of HFD. The results strongly suggest that andrographolide supplementation can be used for prevention and treatment of cardiovascular disease in obese patients.

The Protective Effects of Clams on Hypercholesterolemia in Late-Stage Triple-Transgenic Alzheimer's Diseased Mice Hearts.

To investigate a high cholesterol diet in Alzheimer's disease (AD) mice, they were fed with (2% cholesterol) in five groups with a control group, AD mice group, AD mice plus Meretrix lusoria group, AD mice plus Geloina eros group, and, AD mice plus Corbicula fluminea group for three months, and treated with the fatty acid profiles of clams by gas chromatography (GC). The results showed that treatment with clams for three months reduced Fas/L and Caspase-3 in the Meretrix lusoria and Geloina eros groups, but Fas-associated death domain (FADD) and Caspase-8 were strongly reduced in the Geloina eros group. For the mitochondria-dependent apoptotic pathway, the reduction of apoptosis proteins were observed in the hearts of clams-treated AD mice. BAK and Caspase-9 was reduced in the Meretrix lusoria group, but Caspase-3 and Cytochrome-c were reduced in Geloina eros group. Enhancement of survival proteins p-AKT, p-IGF1R, p-PI3K, Bcl-XL, Bcl2, and the longevity SIRT1 signaling proteins, p-AMPK-α, SIRT1, PGC1-α, p-FOXO3 were observed in clams-treated mice and even more strongly enhanced in the Meretrix lusoria, Geloina eros and Corbicula fluminea groups. This study observed that the ingestion of clams caused a reduction of apoptosis proteins and enhancement of survival and SIRT1 signaling proteins in the hearts.

E2/ER ß Enhances Calcineurin Protein Degradation and PI3K/Akt/MDM2 Signal Transduction to Inhibit ISO-Induced Myocardial Cell Apoptosis.

Secretion of multifunctional estrogen and its receptor has been widely considered as the reason for markedly higher frequency of heart disease in men than in women. 17ß-Estradiol (E2), for instance, has been reported to prevent development of cardiac apoptosis via activation of estrogen receptors (ERs). In addition, protein phosphatase such as protein phosphatase 1 (PP1) and calcineurin (PP2B) are also involved in cardiac hypertrophy and cell apoptosis signaling. However, the mechanism by which E2/ERß suppresses apoptosis is not fully understood, and the role of protein phosphatase in E2/ERß action also needs further investigation. In this study, we observed that E2/ERß inhibited isoproterenol (ISO)-induced myocardial cell apoptosis, cytochrome c release and downstream apoptotic markers. Moreover, we found that E2/ERß blocks ISO-induced apoptosis in H9c2 cells through the enhancement of calcineurin protein degradation through PI3K/Akt/MDM2 signaling pathway. Our results suggest that supplementation with estrogen and/or overexpression of estrogen receptor ß gene may prove to be effective means to treat stress-induced myocardial damage.

Mitotic arrest induced in human DU145 prostate cancer cells in response to KHC-4 treatment.

In this study, the antitumor activity of KHC-4 was analyzed using human prostate cancer (CaP) cells and the underlining anticancer mechanisms of KHC-4 were identified. KHC-4 inhibited cell proliferation and induced cytotoxicity in the castration-resistant CaP DU145 cell line. The most effective concentration of KHC-4 was 0.1 µM. Cell cycle analysis demonstrated that KHC-4 treatment caused G2/M arrest and a subsequent increase in the sub-G1 population. Furthermore, KHC-4 is up-regulated p21, p27, and p53 in a time- and concentration-dependent manner. The exposure of cells to KHC-4 induced Cdk1/cyclin B1 complex activity, which led to cell cycle arrest. Moreover, KHC-4 inhibited the activities of MMP-2 and MMP-9 to inhibit tumor cell metastasis. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1879-1887, 2016.

Helioxanthin suppresses the cross talk of COX-2/PGE2 and EGFR/ERK pathway to inhibit Arecoline-induced Oral Cancer Cell (T28) proliferation and blocks tumor growth in xenografted nude mice.

Helioxanthin, an active compound from Taiwania cryptomerioides Hayata, has been shown to have various biological activities. However, their anticancer effect in oral squamous cell carcinoma has not been well established yet. Helioxanthin inhibited the proliferation of oral squamous cell carcinoma cells in a dose-dependent manner by inducing G2/M phase arrest. Similarly, helioxanthin inhibited cyclooxygenase-2, (COX-2), phosphorylated EGFR, and extracellular-signal-regulated kinases (ERK) protein level and further reduced the nuclear accumulation of phosphorylated epidermal growth factor receptor (pEGFR) and activator protein-1(AP-1) family protein, c-fos. Moreover, helioxanthin at the dose of 20 and 30 mg kg-1 for 15 days reduced the tumor growth in animal model. This study demonstrated that Helioxanthin exerts its anticancer activity against oral cancer cells by downregulating EGFR/ERK/c-fos signaling pathway to inhibit COX-2 level and by activating cyclin-dependent kinase inhibitor (p27) to further induce G2/M cell cycle arrest. This helioxanthin may serve as a novel candidate for oral cancer prevention. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 2045-2056, 2016.

ZAK induces cardiomyocyte hypertrophy and brain natriuretic peptide expression via p38/JNK signaling and GATA4/c-Jun transcriptional factor activation.

Cardiomyocyte hypertrophy is an adaptive response of heart to various stress conditions. During the period of stress accumulation, transition from physiological hypertrophy to pathological hypertrophy results in the promotion of heart failure. Our previous studies found that ZAK, a sterile alpha motif and leucine zipper containing kinase, was highly expressed in infarcted human hearts and demonstrated that overexpression of ZAK induced cardiac hypertrophy. This study evaluates, cellular events associated with the expression of two doxycycline (Dox) inducible Tet-on ZAK expression systems, a Tet-on ZAK WT (wild-type), and a Tet-on ZAK DN (mutant, Dominant-negative form) in H9c2 myoblast cells; Tet-on ZAK WT was found to increase cell size and hypertrophic marker BNP in a dose-dependent manner. To ascertain the mechanism of ZAK-mediated hypertrophy, expression analysis with various inhibitors of the related upstream and downstream proteins was performed. Tet-on ZAK WT expression triggered the p38 and JNK pathway and also activated the expression and nuclear translocation of p-GATA4 and p-c-Jun transcription factors, without the involvement of p-ERK or NFATc3. However, Tet-on ZAK DN showed no effect on the p38 and JNK signaling cascade. The results showed that the inhibitors of JNK1/2 and p38 significantly suppressed ZAK-induced BNP expression. The results show the role of ZAK and/or the ZAK downstream events such as JNK and p38 phosphorylation, c-Jun, and GATA-4 nuclear translocation in cardiac hypertrophy. ZAK and/or the ZAK downstream p38, and JNK pathway could therefore be potential targets to ameliorate cardiac hypertrophy symptoms in ZAK-overexpressed patients.

Attenuation of Magnesium Sulfate on CoCl2-Induced Cell Death by Activating ERK1/2/MAPK and Inhibiting HIF-1α via Mitochondrial Apoptotic Signaling Suppression in a Neuronal Cell Line.

Magnesium sulfate (MgSO4) ameliorates hypoxia/ischemia-induced neuronal apoptosis in a rat model. This study aimed to investigate the mechanisms governing the anti-apoptotic effect of MgSO4 on cobalt chloride (CoCl2)-exposed NB41A3 mouse neuroblastoma cells. MgSO4 increased the viability of NB41A3 cells treated with CoCl2 in a dose-dependent manner. MgSO4 treatment was shown to lead to an increase in the anti-apoptotic Bcl-2 family proteins, with a concomitant decrease in the pro-apoptotic proteins. MgSO4 also attenuated the CoCl2-induced disruption of mitochondrial membrane potential (ΔΨ(m)) and reduced the release of cytochrome c form the mitochondria to the cytosol. Furthermore, exposure to CoCl2 caused activation of the hypoxia-inducible factor 1α (HIF-1α). On the other hand, MgSO4 markedly reduced CoCl2-induced HIF-1α activation and suppressed HIF-1α downstream protein BNIP3. MgSO4 treatment induced ERK1/2 activation and attenuated CoCl2-induced activation of p38 and JNK. Addition of the ERK1/2 inhibitor U0126 significantly reduced the ability of MgSO4 to protect neurons from CoCl2-induced mitochondrial apoptotic events. However, incubation of cultures with the p38 and JNK inhibitors did not significantly affect MgSO4-mediated neuroprotection. MgSO4 appears to suppress CoCl2-induced NB41A3 cell death by activating ERK1/2/ MAPK pathways, which further modulates the role of Bcl-2 family proteins and mitochondria in NB41A3 cells. Our data suggest that MgSO4 may act as a survival factor that preserves mitochondrial integrity and inhibits apoptotic pathways.

Protective effects of Cholestin on ethanol induced oxidative stress in rats.

BACKGROUND: Male Wistar rats were divided into seven groups as follows: group A, basal diet; group B, basal diet with Cholestin at 0.1667 g kg⁻¹ body weight (BW); groups C-F, oral feeding of ethanol at 7.9 g kg⁻¹ BW; groups D-F, Cholestin in diet at 0.1667, 0.3333 and 0.5 g kg⁻¹ BW respectively; group G, silymarin in diet at 200 mg kg⁻¹ BW. RESULTS: The results showed that treatment with Cholestin for 8 weeks reduced the impact of ethanol toxicity on serum markers of liver damage: aspartate aminotransferase (AST), alanine aminotransferase (ALT) and alkaline phosphatase (ALP). The antioxidant system was significantly enhanced: plasma and hepatic thiobarbituric acid-reactive substance (TBARS) levels were lowered while hepatic superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), ethanol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) activities and non-enzymatic antioxidants (vitamin E, vitamin C and GSH) were elevated. CONCLUSION: Cholestin shows a protective effect against hepatotoxicity indices in ethanol-fed rats comparable to that of silymarin, as supported by the evaluation of liver histopathology. The data suggest that Cholestin exerts its hepatoprotective effect by decreasing lipid peroxidation and improving antioxidants status, thus proving itself as an effective antioxidant in ethanol-induced oxidative damage in rats.

Effects of garlic oil on interleukin-6 mediated cardiac hypertrophy in hypercholesterol-fed hamsters.

Hypercholesterol diets are the major causes of cardiac hypertrophy and various cardiac disorders. The purpose of this study is to evaluate the effects of garlic oil on cardiac hypertrophy induced by hypercholesterol diets. Golden Syrian hamsters were fed with 2% cholesterol or 2% cholesterol plus 1% garlic oil for 2 months. Heart architecture changes were measured by hematoxylin-eosin staining and the molecular mechanism was determined by western blotting. Garlic oil reduced whole-heart weight to bone weight ratio, and left ventricle weight to bone weight ratio in the cholesterol-fed group. Moreover, the garlic oil group showed significantly reduced interleukin-6, phosphorylated (p)-extracellular signal-regulated kinase-5, p-mitogen-activated protein kinase-5, calcineurin, nuclear transcription factor of nuclear factor of activated T-cells-3 and p-GATA binding protein 4 when compared with the cholesterol group. However, no changes were observed in gp-130, signal transducer and activator of transcription-3, p-P38 and p-Jun N-terminal kinases protein levels in all groups. The results show that garlic oil may be useful in the treatment of hypertrophy-associated cardiovascular diseases.

Apicidin-resistant HA22T hepatocellular carcinoma cells strongly activated the Wnt/ß-catenin signaling pathway and MMP-2 expression via the IGF-IR/PI3K/Akt signaling pathway enhancing cell metastatic effect.

The IGF-IR/PI3K/Akt signaling pathway inhibited GSK3-ß activity by phosphorylation and this promoted ß-catenin nuclear localization. Our previous study indicated that ß-catenin mRNA level was significantly higher in tumor areas than in non-tumor ones, especially in late pathologic stage tumors. However, ß-catenin inhibition resulted in significantly suppressed migration and invasion ability of HA22T cells. Thus, Wnt/ß-catenin pathway over-activation might be involved in metastatic enhancement of apicidin-resistant HA22T cell metastasis. Apicidin-resistant (AR) HA22T cells showed higher ß-catenin nuclear accumulation and significantly decreased GSK-3-ß protein level, in relation to parental cells. Results also indicated that AR cells increased abundantly in Tbx3, a downstream target of Wnt/ß-catenin that it is implicated in liver cancer. AR cells also inhibited the MEK/ERK/PEA3 pathway which promoted MMP-2 activation. But, apicidin-resistant effect was totally reversed by LY294002 and AG1024. In conclusion, Apicidin-R HA22T cells activated the Wnt/ß-catenin pathway and induced, MMP-2 expression via IGF-IR/PI3K/Akt signaling further enhancing cell the metastatic effects.

A study to evaluate the potential of an in silico approach for predicting dipeptidyl peptidase-IV inhibitory activity in vitro of protein hydrolysates.

A total of 294 edible protein sequences and 5 commercial proteases listed in the BIOPEP database were analyzed in silico. The frequency (A), a parameter in silico described previously, was examined further to calculating the ratio of truncated peptides with Xaa-proline and/or Xaa-alanine to all peptide fragments in a protein hydrolyzed with a protease, using the BIOPEP database. Then the in vitro DPP-IV inhibitory activity was determined using the same 15 protein and protease combinations to evaluate their relationship. The result shows that A values considering the number of Xaa-proline+Xaa-alanine exhibited a strong correlation with in vitro DPP-IV inhibition rates by Pearson's correlation analysis (r=0.6993; P<0.05). Therefore, the in silico approach is effective to predict DPP-IV inhibitory activities in vitro of protein hydrolysates.

Isolation of prolyl endopeptidase inhibitory peptides from a sodium caseinate hydrolysate.

Prolyl endopeptidase (PEP) has been associated with neurodegenerative disorders, and the PEP inhibitors can restore the memory loss caused by amnesic compounds. In this study, we investigated the PEP inhibitory activity of the enzymatic hydrolysates from various food protein sources, and isolated and identified the PEP inhibitory peptides. The hydrolysate obtained from sodium caseinate using bromelain (SC/BML) displayed the highest inhibitory activity of 86.8% at 5 mg mL(-1) in the present study, and its IC50 value against PEP was 0.77 mg mL(-1). The F-5 fraction by RP-HPLC (reversed-phase high performance liquid chromatography) from SC/BML showed the highest PEP inhibition rate of 88.4%, and 9 peptide sequences were identified. The synthetic peptides (1245.63-1787.94 Da) showed dose-dependent inhibition effects on PEP as competitive inhibitors with IC50 values between 29.8 and 650.5 µM. The results suggest that the peptides derived from sodium caseinate have the potential to be PEP inhibitors.

In silico, in vitro and in vivo analyses of dipeptidyl peptidase IV inhibitory activity and the antidiabetic effect of sodium caseinate hydrolysate.

The frequency (A), a novel in silico parameter, was developed by calculating the ratio of the number of truncated peptides with Xaa-proline and Xaa-alanine to all peptide fragments from a protein hydrolyzed with a specific protease. The highest in vitro DPP-IV inhibitory activity (72.7%) was observed in the hydrolysate of sodium caseinate by bromelain (Cas/BRO), and the constituent proteins of bovine casein also had relatively high A values (0.10-0.17) with BRO hydrolysis. 1CBR (the <1 kDa fraction of Cas/BRO) showed the greatest in vitro DPP-IV inhibitory activity of 77.5% and was used for in vivo test by high-fat diet-fed and low-dose streptozotocin-induced diabetic rats. The daily administration of 1CBR for 6 weeks was effective to improve glycaemic control in diabetic rats. The results indicate that the novel in silico method has the potential as a screening tool to predict dietary proteins to generate DPP-IV inhibitory and antidiabetic peptides.

Andrographis paniculata extract attenuates pathological cardiac hypertrophy and apoptosis in high-fat diet fed mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Andrographis paniculata (Burm. f.) Nees (Acanthaceae) has a considerable medicinal reputation in most parts of Asia as a potent medicine in the treatment of Endocrine disorders, inflammation and hypertension. AIM OF THE STUDY: Water extract of A. paniculata and its active constituent andrographolide are known to possess anti-inflammatory and anti-apoptotic effects. Our aim is to identify whether A. paniculata extract could protect myocardial damage in high-fat diet induced obese mice. MATERIALS AND METHODS: The test mice were divided into three groups fed either with normal chow or with high fat diet (obese) or with high fat diet treated with A. paniculata extract (2g/kg/day, through gavage, for a week). RESULTS: We found that the myocardial inflammation pathway related proteins were increased in the obese mouse which potentially contributes to cardiac hypertrophy and myocardial apoptosis. But feeding with A. paniculata extract showed significant inhibition on the effects of high fat diet. CONCLUSION: Our study strongly suggests that supplementation of A. paniculata extract can be used for prevention and treatment of cardiovascular disease in obese patients.

Ameliorative effect of Pracparatum mungo extract on high cholesterol diets in hamsters.

This study was designed to test the lipid-lowering and antioxidative activities of Pracparatum mungo extract (PME), in comparison to its components berberine and glycyrrhizin in cholesterol-fed hamsters. The PME and berberine significantly lowered the atherogenic index compared to glycyrrhizin and the control (P < 0.05). The hepatic HMG-CoA reductase activity was significantly lower in the PME and berberine groups than in the glycyrrhizin group (P < 0.05), while the hepatic ACAT activity was significantly decreased by all treatments with respect to the high cholesterol fed group (P < 0.05). The overall potential of the antioxidant system was significantly enhanced by the PME, berberine and glycyrrhizin supplements as the plasma and hepatic TBARS levels were lowered while the hepatic superoxide dismutase activity and glutathione levels, HMG-CoA reductase, LDL receptor, PPAR, SREBP-2 and CYP7A1 mRNA expressions were increased by the treatments of PME and berberine in comparison with the high cholesterol fed hamsters (P < 0.05). Collectively, these results suggest that the supplementation of PME, berberine and glycyrrhizin increased antioxidant activity in hamsters. Furthermore, we observed that PME and berberine groups promoted the excretion of neutral and acidic sterols (P < 0.05), that could contribute to explain the lower plasma and hepatic cholesterol levels found in the treated animals.

Effect of taurine in chronic alcoholic patients.

A study was undertaken to investigate the dietary effect of taurine in chronic alcoholic patients. The 30 chronic alcoholic patients with 2 to 5 times greater than normal activities of aspartate transaminase (AST) or alanine transaminase (ALT) were selected and equally divided into taurine and control groups. In the taurine group, each patient took 6 g taurine per day, divided into 3 doses, for three months, and then stopped treatment for 1 month. In the control group, patients took a placebo without taurine for 4 months. It was found that the AST and ALT activities and levels of cholesterol, triglyceride (TG), bilirubin, and thiobarbituric acid reactive substances (TBARS) of serum plasma in the taurine group were all decreased, but increased alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) activities and serum vitamins concentrations. Except for the level of TG, all were significantly different after taking taurine for 2 or 3 months. It indicated that taurine plays an important role in the properties of antioxidation and has some improvement on the liver tests of chronic alcoholic patients.

Effects of yam peel extract against carbon tetrachloride-induced hepatotoxicity in rats.

The phenolic acid and flavonoid profiles in yam peel extract were determined by HPLC. Quercetin, hesperidin, and apigenin were predominant components in yam peel extract. Male Wistar rats were orally treated with yam peel extract (100.02, 266.72, and 433.42 mg/kg) or silymarin (200 mg/kg) daily, with administration of CCl4 (1 mL/kg, 20% CCl4 in olive oil) twice a week. Yam peel extract for 8 weeks significantly reduced the impact of CCl4 toxicity on the serum markers of liver damage, aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP). The overall potential of the antioxidant system was significantly enhanced by the yam peel extract supplements as the plasma and hepatic thiobarbituric acid reactive substances (TBARS) levels were lowered, whereas the hepatic superoxide dismutase (SOD) and catalase (CAT) activities and glutathione peroxidase (GSH-Px) protein level were elevated. Yam peel extract decreased the level of nitric oxide (NO) production, tumor necrosis factor-alpha (TNF-α), and nuclear factor-kappa B (NF-κB) in CCl4. These results point out that yam peel extract can inhibit lipid peroxidation, enhance the activities of antioxidant enzymes, and decrease the TNF-α/NF-κB level, nitric oxide production, and inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expressions. Therefore, it was speculated that yam peel extract protects rats from liver damage through its anti-inflammation capacity.

Effect of cholestin on toxicity of vitamin A in rats.

A study was undertaken to investigate the effect of cholestin on the toxicity of vitamin A in male wistar rats. The rats were divided into six groups and fed different diets with or without supplement of 1% cholestin and 25,000-50,000 (IU) vitamin A for 2 months. Hence, the symptoms of vitamin A toxicity in rats included loss of body weight, hepatotoxicity and nephrotoxicity. However, these toxic effects of vitamin A were significantly reduced when the rats fed a diet supplemented with cholestin. Furthermore, the level of vitamin A in the serum of rats treated with cholestin and vitamin A was higher than that of the rats treated with vitamin A alone. It indicated that cholestin might play a role in reducing the toxic effect of vitamin A in rats.

Dietary taurine reduces zinc-induced toxicity in male Wistar rats.

Taurine is an agent for treating the heavy metal intoxication and presence of metals such as zinc, copper, and iron may have a role in heavy metal toxicity, a study was undertaken to investigate the effect of taurine on the toxicity of zinc in male Wistar rats. The rats were divided into 8 groups and fed different diets with or without supplement of 5% taurine and 150 to 600 ppm zinc for 2 mo. It was found that the body weight of rats, the ratios of liver and kidney weight to body weight, and the level of glutathione in the liver were decreased with increasing the dose of zinc. The levels of zinc in the liver, kidney, and plasma, the levels of malondialdehyde in the plasma, the levels of thiobarbiture acid-reactive substances in the liver, the activities of aspartate transaminase, alanine transaminase in the plasma, the levels of blood urea nitrogen and creatinine in the plasma of rats were increased with the increasing dose of zinc. Hence, symptoms of zinc toxicity in rats included loss of body weight, hepatotoxicity, and nephrotoxicity. However, these toxic effects of zinc were significantly reduced when the rats fed diet with supplement of taurine. Furthermore, the level of zinc in the feces of rats treated with taurine and zinc was higher than that of rats treated with zinc alone. It indicated that taurine thereby leading to a decreased absorption of dietary zinc and promoted excretion.