Shikonin inhibits neuronal apoptosis induced by OGD through targeting PI3K/Akt signaling pathway

AIM: To explore the effect of shikonin on rat primary cortical neurons in oxygen-glucose deprivation (OGD)-induced injury model.METHODS: The neurons were pretreated with shikonin at different concentrations (0.02, 0.2, 2 and 20 μmol/L) followed by treatment with OGD. Lactate dehydrogenase (LDH) release assay and fluorescein diacetate/propidium iodide (FDA/PI) double staining were used to detect neuronal viability and apoptosis, and then the optimal concentration of shikonin was determined. LY294002 (PI3K/Akt signaling pathway inhibitor, 1 μmol/L) was added before the addition of shikonin, and the protein level of p-Akt (Ser473) in the neurons was determined by Wes-tern blot. LDH release assay and FDA/PI double staining were also used to detect neuronal viability and apoptosis.RESULTS: A certain concentration (0.2~20 μmol/L) of shikonin increased the viability of impaired neurons (P<0.05) and the protein level of p-Akt (Ser473) in the neurons (P<0.05). The effect of shikonin on neuronal p-Akt (Ser473) levels and the cell death were blocked by LY294002 (P<0.05).CONCLUSION: A certain concentration of shikonin reduces OGD-induced apoptosis of rat primary cortical neurons by activating PI3K/Akt signaling pathway.     

CUDC-907 induces DNA damage and autophagy in glioma cells

AIM:To investigate the effect of CUDC-907, a dual histone deacetylase (HDAC) and phosphatidylinositol 3-kinase (PI3K) inhibitor, on the DNA damage, cell cycle distribution and autophagy in human glioma U251 cells. METHODS:U251 cells were treated with CUDC-907 of different concentrations, and the cell viability was detected by MTT assay. The quantitative γ-H2AX foci were determined by laser scanning confocal microscopy. The cell cycle distribution of U251 cells was examined by flow cytometry. The protein expression was determined by Western blot analysis. RESULTS:CUDC-907 inhibited the cell viability and the phosphorylation of Akt and p70 ribosomal protein S6 kinase (p70s6K) in the U251 cells (P<0.05). In CUDC-907-treated cells, the number of γ-H2AX foci and protein expression of γ-H2AX were increased significantly (P<0.05). CUDC-907 also induced cell arrest in the G₂/M phase by up-regulating the expression of p21, and inhibiting the protein level of cyclin B1 and the phosphorylation of cell division cycle protein 2 (Cdc2). In addition, CUDC-907 triggered cell autophagy, and inhibition of autophagy increased CUDC-907-induced DNA damage of U251 cells. CONCLUSION:CUDC-907 significantly inhibits PI3K/Akt signaling pathway, induces DNA damage and arrests cell cycle in G₂/M phase. Blockage of autophagy promotes CUDC-907-induced DNA damage of U251 cells.     

Effect of FGFR1 expression knock-down on biological characteristics of infantile hemangioma endothelial cells

AIM: To study the effect of fibroblast growth factor receptor 1 (FGFR1) expression knock-down on the viability, apoptosis, invasion and migration of infantile hemangioma endothelial cells (HemECs). METHODS: FGFR1 was down-regulated by FGFR1 small interfering RNA (si-FGFR1) transfection. The viability of the cells was measured by CCK-8 assay. The apoptotic rate was analyzed by flow cytometry and the invasion and migration abilities were determined by Transwell assay. The protein levels of phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), and phosphorylated AKT (p-AKT) were examined by Western blot. RESULTS: Transfection of si-FGFR1 into HemECs had significant effects on inhibiting cell viability (P<0.05), promoting apoptosis (P<0.05), and decreasing cell invasion and migration abilities (P<0.05). The results of Western blot showed that knockdown of FGFR1 gene expression in the cells reduced the protein levels of PI3K and p-AKT (P<0.05), and had no significant effect on AKT protein level. CONCLUSION: Knock-down of FGFR1 expression changes the biological characteristics of endothelial cells in infantile hemangiomas by regulating PI3K/AKT signaling pathway.     

Research progress on autophagy in treatment of glioma

Autophagy is a transport pathway from the cytoplasm to the lysosome, which is a major intracellular degradation/recycling system ubiquitous in eukaryotic cells. Autophagy regulation has achieved some gratifying results in the treatment of glioma. It is currently an exciting field of clinical development. In chemotherapy or radiotherapy, autophagy-related drugs are currently used in vitro and in vivo for treating tumors with significant effects. Autophagy inducers and inhibitors may potentially block tumor formation and enhance the anti-cancer immune response. A more comprehensive understanding of the role of autophagy in different stages of glioma development may guide the development of new therapeutic strategies.     

Cepharanthine induces autophagy via PI3K/AKT/mTOR signaling pathway in ovarian cancer SKOV3 cells

AIM: To investigate the autophagy of human ovarian cancer SKOV3 cells induced by cepharanthine and to explore its mechanism. METHODS: The effect of cepharanthine on the viability of ovarian cancer SKOV3 cells was measured by CCK-8 assay. The SKOV3 cells were treated with cepharanthine, and then the formation of autophagosome was observed with acridine orange staining under fluorescence microscope. The protein levels of LC3, AKT, p-AKT, mTOR, p-mTOR and GAPDH in the SKOV3 cells treated with cepharanthine were determined by Western blot.RESULTS: Cepharanthine significantly inhibited the viability of ovarian cancer SKOV3 cells in a dose-dependent manner (P<0.05). The number of the intracellular acidic autophagosomes with bright red fluorescence was significantly increased after cepharanthine treatment in the SKOV3 cells. The expression of LC3-Ⅱ in SKOV3 cells was significantly enhanced after cepharanthine treatment. Furthermore, treatment with cepharanthine in the SKOV3 cells also resulted in a significant down-regulation of phosphorylated form of AKT and mTOR (P<0.01), while the total protein level was not changed. Combination of cepharanthine and 3-methyladenine resulted in a substantial decrease in the cell viability compared with using cepharanthine alone.CONCLUSION: Cepharanthine significantly inhibits the growth of human ovarian cancer SKOV3 cells and induces the autophagy, which may be correlated with down-regulation of PI3K/AKT/mTOR signaling pathway.     

水稻苗期根系铵钾离子吸收的交互作用研究

【目的】为探讨水稻幼苗根系NH_4~+、K~+吸收的交互作用,深化水稻养分吸收理论,【方法】采用溶液培养的方法,对低钾及高钾浓度下水稻在有铵和无铵时的K~+吸收动力学特征进行了研究,对不同钾浓度下水稻根系NH_4~+的吸收速率进行了比较。【结果】1)当K~+0.2 mmol/L时,水稻根系通过高亲和转运系统吸收K~+服从Michaelich-Menten动力学方程;NH_4~+的存在显著降低K~+的最大吸收速率(Vmax),且降幅随着NH_4~+浓度的增加而增大;NH_4~+对水稻根表载体与K~+的亲和力(Km)影响较小,在1.62 mmol/L NH_4~+浓度下,水稻品种齐粒丝苗和沪科3号的Km分别下降了12.33%和16.46%,远低于Vmax 47.30%和39.21%的降幅。2)当K~+0.5 mmol/L时,水稻根系K~+低亲和转运系统发挥作用,K~+吸收速率随浓度的增加而不断增加,呈不饱和特征;但在相同K~+浓度下,水稻根系的K~+吸收速率随NH_4~+浓度的增加而下降。3)水稻根系对NH_4~+的吸收速率随着NH_4~+浓度的增加而增加;在相同NH_4~+浓度下,水稻根系对NH_4~+的吸收速率受K~+浓度的影响很小。【结论】NH_4~+抑制水稻苗期根系K~+的高亲和转运和低亲和转运,NH_4~+对K~+高亲和吸收的影响主要是由于铵竞争细胞膜上的钾载体所致;外界K~+浓度的变化对水稻幼苗的NH_4~+吸收速率影响很小。水稻铵钾的交互作用主要表现在NH_4~+对K~+吸收的抑制作用。     

Potassium fertilization improves apple fruit (Malus domestica Borkh. Cv. Fuji) development by regulating trehalose metabolism

Potassium (K) fertilization and antioxidant enzymes both positively influence plant growth and development. However, it is not known whether K treatment improves fruit development via increasing soluble sugar. In this study, K-treated apple (Malus domestica Borkh.) fruit was harvested from 60 to 150 days after full bloom and was analyzed for ROS production and trehalose metabolism. The results show that K fertilization improved fruit firmness, increased growth according to several parameters, increased soluble sugar content, and decreased ROS production. The ascorbate metabolic pathway more effectively reduced ROS production than catalase and peroxidase (POD) did under K treatment. Trehalose-treated fruit also showed higher activity of ascorbate-related enzymes (DHAR, GR, and APX) compared with non-treated fruit. The changes of antioxidant enzyme activity in trehalose-treated fruit corresponded to those in K-treated fruit. Moreover, trehalase (TREH) activity in fruit was notably reduced by K treatment. This demonstrates that K influences ROS production via regulating trehalose content and TREH activity in fruit. This study provides new insight into the K mechanism which improves fruit development, including fruit firmness and size.     

追施不同形态氮肥对不同钾效率基因型棉花生长及产量品质的影响

【目的】探讨高、低供钾水平下,追施氮肥形态对棉花生长、钾素吸收利用以及产量、品质的影响。【方法】选择钾高效棉花品种辽棉18、冀棉958和钾低效棉花品种新棉99B为材料,进行营养钵培养试验,设置38.01mg·kg~(-1)和152.24 mg·kg~(-1)两个供钾水平,追施铵态氮肥(硫酸铵)和硝态氮肥(硝酸钙)两种形态氮素肥料。【结果】供钾不足会降低棉花果枝始节和单株成铃数,高钾处理棉花干物质积累量、钾累积量、钾利用指数以及产量显著高于低钾处理;与追施硝态氮肥相比,追施铵态氮肥会降低棉株高度、果枝数和单株成铃数,减少籽棉产量和总干物质积累量;追施铵态氮肥处理棉花对钾素的吸收和利用显著低于追施硝态氮肥处理。【结论】钾低效基因型品种棉花在钾素供应不足时对追施铵态氮肥更敏感,且棉花成熟越晚受到追施肥料氮素形态影响越大。     

Mechanism of quercetin improving rat coronary artery myogenic response under high glucose

AIM: To investigate the mechanism of quercetin improving rat coronary artery myogenic response under high glucose (HG) by measuring muscle tension of coronary arterial ring and recording voltage-gated K⁺ channel (Kv) current of coronary artery smooth muscle cells by whole cell patch clamp. METHODS: The coronary rings from the normal SD rats were acutely isolated, and then divided into 6 groups: (1) control group; (2) HG group; (3) HG+low dose (3 μmol/L) of quercetin group; (4) HG+moderate dose (10 μmol/L) of quercetin group; (5) HG+high dose (30 μmol/L) of quercetin group; (6) HG+C6303 (PKC inhibitor)+high dose of quercetin group. Determinations of coronary artery response to vasoconstrictor (60 mmol/L KCl or 0.1 mmol/L U46619) or vasodilator (ACh at 10^-9~10^-5 mol/L) were performed, and the percentage of coronary ring tension was calculated using the contraction as 100% caused by 60 mmol/L KCl. The rat coronary artery smooth muscle cells were acutely isolated for recording the Kv current using whole cell patch clamp. RESULTS: Compared with control group, the contraction amplitudes to 60 mmol/L KCl or 0.1 mmol/L U46619 were significantly increased under HG incubation. Quercetin intervention concentration-dependently reduced the coronary artery contraction amplitude. Incubation of PKC specific inhibitor C6303 attenuated the effect of quercetin. Compared with control group, the diastolic amplitude to ACh decreased significantly in HG group, and quercetin intervention concentration-dependently increased the coronary artery diastolic amplitude. Incubation of PKC specific inhibitor C6303 attenuated the effect of quercetin. Compared with control group, HG incubation inhibited Kv current of coronary artery vascular smooth muscle cells significantly, and quercetin intervention attenuated the inhibitory effect of HG on Kv current intensity. Incubation of PKC specific inhibitor C6303 attenuated the effect of quercetin. CONCLUSION: Quercetin has a protective effect on myogenic response of coronary artery under HG and the effects is related to the increase in Kv current and the activation of PKC in vascular smooth muscle cells.