The protection of selenium on cadmium-induced inhibition of spermatogenesis via activating testosterone synthesis in mice.

Selenium (Se) is an essential trance element in testis. However, the potential protective effects of Se against cadmium (Cd)-induced reproductive toxicity remained to be elucidated. Male ICR mice were orally administered by gavage with Na2SeO3 (0.1, 0.2, 0.4 mg/kg BW) for 1h prior to CdCl2 (5 mg/kg BW) alone or in combination for 15, 25 or 35 days. Cd exposure caused a significant decrease in body weight, sperm concentration and motility as well as plasma testosterone level which was accompanied by decreased antioxidant enzymatic activity of SOD and GSH-Px and by increased lipid peroxidation (as malondialdehyde, MDA). Se pretreatment compensated deficits in the sperm parameters (concentration, motility and morphology) induced by Cd. Se (0.4 mg/kg BW) treatment significantly increased serum testosterone level that was reduced by Cd (on 15th, 25th and 35th day) (P<0.01). Se treatment ameliorated Cd-induced reduction in testicular steroidogenic acute regulatory (StAR) and 17ß-hydroxysteroid dehydrogenase (17ß-HSD) activities. The present study suggest that the protective potential of Se against Cd-induced reprotoxicity might be due to up-regulation StAR and testosterone synthetic enzyme activity, which could be useful for increasing testosterone synthesis for achieving optimum protection in sperm quality and spermatogenesis.

Liquiritigenin inhibits serum-induced HIF-1α and VEGF expression via the AKT/mTOR-p70S6K signalling pathway in HeLa cells.

Liquiritigenin (LQ) is a non-toxic dietary flavonoid with chemopreventive and anticancer properties. However, the mechanism of its antiangiogenesis remains unclear. Hypoxia-inducible factor-1α (HIF-1α) and its downstream target, vascular endothelial growth factor (VEGF), play a critical role in tumour angiogenesis and represent an attractive chemotherapeutic target. In this study, we investigated the effect of LQ on the molecular mechanism of angiogenesis. We found that LQ inhibited VEGF expression at both mRNA and protein levels. Liquiritigenin did not affect HIF-1α expression at the mRNA level, but it dramatically inhibited both serum- and mimicked hypoxic-induced HIF-1α protein accumulation in HeLa cells. Furthermore, we showed that LQ inhibited serum-induced expression of HIF-1α by reducing its stability and decreased the synthesis in a dose-dependent manner. Mechanistically, we demonstrated that LQ inhibited HIF-1α and VEGF expression involved in blocking the protein kinase B (PKB/Akt) signalling pathway, and the mechanisms correlated with dephosphorylation of the mammalian target of rapamycin (mTOR) and its effector ribosomal protein S6 kinase (p70S6K). In addition, LQ inhibited VEGF-induced formation of capillary-like structures in human umbilical vein endothelial cells (HUVEC). Taken together, our study provided valuable insights into the mechanism of antiangiogenic effect of LQ.

Effect of Wujijing Oral Liquid on menstrual disturbance of women.

OBJECTIVE: Wujijing Oral Liquid (WJJ) contained principally the flesh essence of the black-boned chicken. As a kind of food and medicine in China, it was used to treat the menstrual disturbance traditionally, but the exact mechanism of the action was not yet clear. The clinical effects of the WJJ on the symptoms of the menstrual disturbance and the reproductive hormones were studied in this paper. MATERIALS AND METHODS: The 53 women with the menstrual disturbance were selected as the study object, and then they were randomly divided to receive either WJJ 10mL twice daily (n=28) or the placebo (n=25) from the 1st day after menstrual flow for 2 menstrual cycles. On the 1st day after the discontinuation of the medication but before the treatment, the scores for the menstrual pattern and the related symptoms were obtained and the blood samples were collected to test the reproductive hormones. The serum levels of the follicle-stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL) and estradiol (E2) were examined by enzyme-linked immunosorbent assay (ELISA). The levels of progesterone (P) and testosterone (T) in serum were measured by the radioimmunoassay. RESULTS: The score for the primary and related symptoms of the menstruation was increased significantly among patients treated with the WJJ. The differences on the FSH, PRL, and E2 levels of patients were significant before and after the treatment with WJJ. Comparing the WJJ group and the placebo group, the levels of P and T differed significantly after treatment. The oral liquid of WJJ was found to be safe, as it did not cause any change in the hepatic and renal functional parameters. CONCLUSION: The oral liquid of Wujijing could improve the menstrual disturbance and were generally safe and well tolerated. The possible mechanism could be associated with its effects in reinforcing the kidney and regulating the hypothalamus-pituitary-ovary axis (HPOA).

The protection of selenium on ROS mediated-apoptosis by mitochondria dysfunction in cadmium-induced LLC-PK(1) cells.

Selenium, an essential trace element, showed the significant protective effects against liver and kidney damage induced by some heavy metals. However, the mechanism how selenium suppresses cadmium (Cd)-induced cytotoxicity remains unclear. In this study, we investigated the protective mechanism of selenium on Cd-induced apoptosis in LLC-PK(1) cells via reactive oxygen species (ROS) and mitochondria linked signal pathway. Studies of PI and Annexin V dual staining analysis demonstrated that 20 microM Cd-induced apoptosis as early as 18 h. A concomitant by the generation of ROS, the loss of mitochondrial membrane potential, cytochrome c (cyt c) release, activation of caspase-9, -3 and regulation of Bcl-2 and Bax were observed. N-acetylcysteine (NAC, 500 microM), a free radical scavenger, was used to determine the involvement of ROS in Cd-induced apoptosis. During the process, selenium played the same role as NAC. The anti-apoptosis exerted by selenium involved the blocking of Cd-induced ROS generation, the inhibition of Cd-induced mitochondrial membrane potential collapse, the prevention of cyt c release, subsequent inhibition of caspase activation and the changed level of Bcl-2 and Bax. Taken together, we concluded that Cd-induced apoptosis was mediated by oxidative stress and selenium produced a significant protection against Cd-induced apoptosis in LLC-PK(1) via ameliorating the mitochondrial dysfunction.

Effect of liquiritigenin, a flavanone existed from Radix glycyrrhizae on pro-apoptotic in SMMC-7721 cells.

Liquiritigenin is a flavanone existed in Radix glycyrrhizae. The objective of this study is to explore the effects of liquiritigenin on SMMC-7721 cells and its possible mechanism. The viability of liquiritigenin treat cells was decreased in a dose-dependent manner assayed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenylterazolium bromide (MTT), and apoptotic morphological changes also be observed, such as chromatin condensation and nuclear fragmentation. Assessment of apoptotic cells by flow cytometry indicated that cells fell into apoptosis after 0.4mM liquiritigenin treatment. In addition, a concomitant time-dependent increase in caspase-3 activity was also observed. The level of p53 protein increased and Bcl-2 protein decreased time-dependently. Further studies found the induction of apoptosis by liquiritigenin was accompanied with the production of reactive oxygen species (ROS), disruption of mitochondrial membrane potential and depletion of antioxidant enzymes. The significant ROS generation was firstly found at 3h and being time-dependent until 9h. A time-dependent decrease in membrane potential occurred, and significant loss appeared at 9h and 12h. Furthermore, pretreatment of N-acetyl-cysteine (NAC), ROS production and apoptosis induced by liquiritigenin were both suppressed. In sum, this paper indicated the cytotoxicity of liquiritigenin on SMMC-7721 cells may via effect on generation of ROS, later lead to cell apoptosis.

[Determination of inorganic elements in rat serum, and vegetable and fruit ferment liquid by ICP-MS].

In the present paper, the contents of thirteen inorganic elements in rat serum, and vegetable and fruit ferment liquid (VFFL) were measured by ICP-MS in order to study the anti-tumor effect of VFFL. Serum or VFFL was digested in nitric and perchloric acids at room temperature and then heated until dryness. The residue was dissolved with 1% (phi) nitric acid prior to ICP-MS analysis. The element contents were quantitated by using 45Sc, 103Rh and 187Re as the internal standards, respectively, according to the rule of close mass number. Certificate references bovine serum (GBW(E)090006) and tea (GBW070605) were employed to validate the proposed method, and the analysis results of most elements in two certificate references were in agreement with their reference values. The intra-day and inter-day precisions of the method in terms of relative standard deviation (RSD) were mainly below 10% and below 15%, respectively. The spiked recoveries for most of studied elements were 80%-110% in rat serum and 90%-120% in VFFL. This method was rapid, highly sensitive, and especially suitable to being applied to small quantity of biological samples with greatly different elements contents. Therefore, we measured the content of thirteen elements in the sera of rats, where in were induced liver cancer by revulsant, and the rate were fed with different dosage of VFFL in intragastric infusion at the same time. It was preliminarily found that the concentrations of some elements in sera of different experiment groups of rats were significantly different, implying the potential anti-tumor effects of VFFL.

Cytoprotective effects of selenium on cadmium-induced LLC-PK1 cells apoptosis by activating JNK pathway.

Extensive studies have indicated that the apoptosis pathway appears to be associated with intracellular reactive oxygen species (ROS) production in cadmium-induced nephrotoxicity, however, the precise cellular mechanism remains unclear. The purpose of this study was to determine the relationships between the activation of phosphorylated c-jun N-terminal kinase (JNK) and cadmium-induced apoptosis, and assess the possible cytoprotective mechanism of selenium. Our study clearly revealed cadmium treatment caused apoptosis in LLC-PK1 cells, which was partially suppressed by pretreatment with selenium, an antioxidant nutrient. Further studies found the phosphorylation of JNK kinase increased with exposure to cadmium for 3 h, even remained elevated at 9 h in the time course study, and the activation of phosphorylated JNK was detected in a dose-dependent manner. In addition, a concomitant time-dependent increase in caspase-3 activities was observed by cadmium treatment. During the process, selenium played the same role as N-acetyl-L-cysteine (NAC), a free radical scavenger. Pretreatment of cells with selenium partially suppressed of the phosphorylation of JNK, coupled with caspase-3 activation involved in cadmium-induced apoptosis. In conclusion, our studies provided a molecular linkage between the phosphorylation of JNK and cadmium-induced LLC-PK1 cells apoptosis, and demonstrated selenium also contributed a potentially protection to prevent cadmium-cytotoxicity.

Inhibitory effects of vegetable and fruit ferment liquid on tumor growth in Hepatoma-22 inoculation model.

The aim of this study was to determine the anti-tumor effect of vegetables and fruits ferment liquid (VFFL) in human hepatoma-22(H22)-bearing mice. Mice bearing H22 were randomly divided into four groups, that is a control group and three VFFL groups (16.7, 33.3 and 66.6 ml/kg). Inhibition rates of tumor, thymus and spleen index were observed. The apoptosis was analyzed by flow cytometry and the apoptotic body was observed under an electron microscope. A survival study was performed on the same model for the duration of 60 days. For this survival study, the mice were divided into five groups, which included a control group, three VFFL groups (16.7, 33.3 and 66.6 ml/kg) and a CP group. Tumor inhibition rates for VFFL16.7, 33.3 and 66.6ml/kg were 25.7%, 35.0 % (p<0.05) and 49.1 % (p<0.01) respectively at 30d, increasing in proportion to the concentration of VFFL given. Thymus and spleen indices of the VFFL groups were also higher than that of the control group. The apoptotic rates in VFFL 16.7, 33.3 and 66.6 ml/kg groups were 20.5%, 24.0% and 15.8% respectively, while it was only 6.82% in control group. In particular, the apoptotic body in the 66.6 ml/kg group exhibited typical apoptotic characteristics, e.g., condensation of nucleus, chromatin fragmentation, and shrinkage of cytoplasm. For the survival study, the mice in the VFFL 66.6ml/kg group exhibited significantly extended survival rates compared with the mice in the control group (p<0.05). This study concludes that VFFL possesses anti-tumor properties, which it exhibits by inducing apoptosis and prolonging life in H22 tumor-bearing mice.