Zerumbone attenuates MPP+-induced cytotoxicity in human neuroblastoma SH-SY5Y cells by inhibition of oxidative stress

AIM: To investigate the role of zerumbone (ZER) in 1-methyl-4-phenylpyridinium (MPP⁺)-induced cytotoxicity of human neuroblastoma SH-SY5Y cells. METHODS: Human neuroblastoma SH-SY5Y cells were cultured in vitro and the protective effect of ZER against MPP⁺-induced cytotoxicity was measured by CCK-8 assay. Flow cytometry was used to determine the apoptosis and reactive oxygen species (ROS). The expression of Parkinson disease protein 7 (PARK7) was knocked-down by using PARK7-specific short hairpin RNA (shRNA). The protein levels of PARK7, nuclear factor E2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) were determined by Western blot. RESULTS: MMP⁺ remarkably reduced the cell viability in a dose-dependent and time-dependent manner. The SH-SY5Y cell injury model was established by treatment with MPP⁺ at 600 μmol/L for 24 h. ZER up-regulated the protein levels of PARK7 and Nrf2 (P<0.05), alleviated apoptosis (P<0.05), and reduced ROS production (P<0.05) in the SH-SY5Y cell injury model. Meanwhile, N-acetyl-L-cysteine (NAC) had the similar functions. Moreover, significant reductions in the protein levels of Nrf2 and HO-1 (P<0.05), and obvious increases in apoptosis (P<0.05) and ROS level (P<0.05) were demonstrated in PARK7-knockdown cells. CONCLUSION: ZER protects SH-SY5Y cells against MPP⁺-induced cytotoxi-city, which may be related to activation of PARK7/Nrf2/HO-1 pathway, and subsequent attenuation of oxidative stress and apoptosis.     

Cell direct reprogramming：a new technique for treating diabetes

Diabetes is characterized by an absolute or relative deficiency in β-cell mass, which cannot be reversed with existing therapeutic strategies. The restoration of the endogenous islet β-cells can stabilize the level of blood glucose. The islet β-cells can be obtained from the directional differentiation of stem cells, but the process is complex and has the risk of teratomas generation. Cell direct reprogramming, one terminal differentiated cell can transdifferentiate into another kind of terminal differentiated cell, which is other than directional differentiation from stem cells. Direct reprogramming gives rise to the generation of islet β-cells from one terminal differentiated cell, may be preferable for diabetes therapy because of its unique advantage.     

Dexmedetomidine attenuates acute alcoholic hepatic injury in mice

AIM: To investigate the effects of dexmedetomidine (DEX) on acute alcoholic hepatic injury in mice and to explore the possible mechanisms. METHODS: Kunming mice (n=50) were randomly divided into 5 groups (n=10): normal saline control (NS) group, acute alcoholic hepatic injury model (E) group, low-dose (10 μg/kg) DEX (E+L) group, medium-dose (50 μg/kg) DEX (E+M) group and high-dose (100 μg/kg) DEX (E+H) group. The animals were sacrificed at 6 h after gavage of alcohol or normal saline. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), malondialdehyde (MDA), glutathione (GSH) and superoxide dismutase (SOD) were measured. The livers were removed for evaluation of histological characteristics and determining the content of tumor necrosis factor-α (TNF-α) amd interleukin-1β (IL-1β) in the liver tissues by ELISA. The expression levels of cytochrome P450 2E1 (CYP2E1) and nuclear factor-κB (NF-κB) in the liver tissues were evaluated by Western blot. RESULTS: Compared with NS group, the levels of ALT, AST and TG were obviously increased in E group, which were obviously decreased in E+M and E+H groups. Compared with NS group, the levels of TNF-α, IL-1β and MDA were obviously increase in E group, which were obviously decreased in E+M and E+H groups. Compared with NS group, the activity of SOD and the content of GSH were obviously decreased in E group, which were obviously increased in E+M and E+H groups. Compared with NS group, the expression of CYP2E1 and NF-κB was obviously increase in E group, which was obviously decreased in E+M and E+H groups. Compared with NS group, ethanol induced marked liver histological injury, which was less pronounced in E+M and E+H groups. CONCLUSION: DEX has a protective effect on mouse liver with acute alcoholic injury by the involvement in the processes of antioxidation and antiinflammation, and its mechanism may be associated with the inhibition of CYP2E1 and NF-κB expression.     

Cardioprotective effect of sevoflurane preconditioning on ROS production in rat heart with ischemia-reperfusion

AIM: To investigate the effects of sevoflurane preconditioning on the production of reactive oxygen species (ROS) and nitric oxide (NO), the activity of superoxide dismutase(SOD), glutathione peroxide(GPx) and catalase(CAT) in myocardial ischemia-reperfusion injury(IRI). METHODS: Sixty rats were randomly allocated to 8 groups. Following 2% sevoflurane preconditioning for 30 min, the left anterior descending artery was ligated for 30 min and then reperfused for 120 min in vivo. The infarction size of the hearts was measured with the staining of 2,3,5-triphenyltetrazoliumchloride. The myocardial apoptotic index was measured by the method of TUNEL. The ROS fluorescent probe dihydroethidium was used for the measurement of ROS. The myocardium was homogenized for the measurement of NO, SOD, GPx and CAT. To evaluate the effects of ROS and NO on the cardioprotection of sevoflurane preconditioning, ROS scavenger N-(2-mercaptopropionyl) glycine (2-MPG) or NOS inhibitor N^ω-nitro-L-arginine methyl ester (L-NAME) were employed to block their actions. RESULTS: Compared with control group, the production of ROS was induced by sevoflurane preconditioning before ischemia-reperfusion injury (12.0±0.8 vs 2.6±0.5, P<0.05) and decreased after ischemia-reperfusion injury (16.2 ±0.9 vs 24.9±1.3, P<0.05). 2-MPG decreased the elevation of ROS caused by sevoflurane preconditioning before ischemia-reperfusion injury (5.1±0.7 vs 12.0±0.8, P<0.05). No difference of ROS production between treating with 2-MPG+Sevo+IRI and with IRI (24.9±1.4 vs 24.9±1.3, P>0.05) was observed. Compared with control group, sevoflurane preconditioning also induced the generation of NO (34.5±3.2 vs 15.9±1.4, P<0.05) and the activity of SOD(1.5±0.5 vs 0.6±0.2, P<0.05), GPx(22.8±2.5 vs 12.7±2.2, P<0.05) and CAT(15.5±1.8 vs 11.2±1.4, P<0.05). 2-MPG blocked the increase in NO production and inhibited the activity of SOD,GPx,CAT. L-NAME also attenuated the activity of SOD,GPx,CAT. CONCLUSION: Sevoflurane preconditioning protects the rat heart against ischemia-reperfusion injury by reducing the infarction size and apoptosis. Production of ROS at sub-injury dose induced by sevoflurane preconditioning stimulates the myocardium to create SOD,GPx,CAT and NO, thus inhibiting the further formation of ROS and protecting the heart under the condition of ischemia-reperfusion.     

Effects of Chemical Components on the Amount of Green Tea Cream

The relationship between the amount of tea cream and the chemical components contents in the original green tea infusion was investigated. The results showed that the amounts of tea cream in the green tea infusions obtained from different cultivars and different parts of new shoots were varied. There were many chemical components participating in the formation of green tea cream. However, there were only the contents of caffeine (Y=0.85, P＜0.01) and polyphenols (Y=0.65,P＜0.05) in the original green tea infusion highly correlated with the amount of green tea cream. Stepwise regression analysis of overall chemical components indicated that the contents of caffeine and gallated catechins in the original green tea infusion had a significant effect (P＜0.0 1) on green tea cream levels. Cream (g L-1)=-172.071+ 0.129×Ccaffeine+0.024×Ggallated catechins (R2=0.936). The amount of green tea cream can be predicted by the contents of gallated catechins and caffeine in the original tea infusion. Principal component analysis also indicated that catechins, minerals, and polysaccharides were the important chemical components in the formation of green tea cream.     

Legumain对TNF-α和CHX共同诱导的细胞凋亡的拮抗作用

目的探讨Legumain（LGMN）对肿瘤坏死因子-α（TNF-α）和放线菌酮（CHX）共同诱导的细胞凋亡的抑制作用,为进一步研究Legumain在肿瘤细胞生长增殖的分子机制以及相关的肿瘤治疗提供实验基础和理论依据。方法采用Western Blot和水解多肽发色底物法分别测定人胚胎肾（HEK）293细胞株和小鼠成纤维细胞L929细胞株中的Legumain表达量,细胞增殖实验（MTT法）检测Legumain对由TNF-α和CHX共同介导的细胞增殖抑制的拮抗作用,激光共聚焦显微镜和流式细胞技术观察Legumain的抑制细胞凋亡效果。结果实验组中的HEK293＋Legumain细胞和采用AEPI预先处理的L929细胞对一定浓度范围的TNF-α和CHX共同介导的细胞增殖抑制有的拮抗作用（P＜0.01或0.05）;激光共聚焦显微镜观察,Annexin V-PI双染法结果表明实验组的HEK293＋Legumain细胞未发生细胞凋亡现象;流式细胞技术,JC-1染色法测定细胞线粒体膜电位结果表明实验组的HEK293＋legumain细胞的凋亡百分比明显低于对照组（＜0.01）。结论Legumain具有拮抗由TNF-α和CHX共同诱导的细胞凋亡作用。     

Effect of integrin α2 on adhesion of neuroblastoma cells to co llagen

AIM:To study the effect of integrin α2 on adhesion of SK-N-SH neuroblastoma cells to collagen.METHODS:Adhesion of the SK-N-SH cells to immobilized collagen w as tested with various concentration of Mg2+,Ca2+ and with 10 μg/L anti-α2 monoclonal antibody (mAb) 6F1.A570 was detected as adhesio n cell numbers.RESULTS:Mg2+-dependent adhesion of SK-N-SH cells to type I collagen was increased significantly,with peak adhesion at concentration of 1 mmol/L Mg2+.A570 with or without Mg2+ was 0.59±0.03 and 0.25±0.01 respectively (P<0.01).No effect of Ca2+ was found on SK-N-SH cell adhesion.6F1 (10 mg/L) blocked the adhesion completely.CONCLUSION:Integrin α2 regulates neuroblastoma cells adhesion t o collagen and suggest that it may play a role in tumor cell migration,invasion and metastasis.     

Effects of TRAF3 on inhibiting cyst formation induced by NF-κB signaling in kidney

AIM: To investigate the effects of tumor necrosis factor (TNF) receptor associated factor 3 (TRAF3) on the signaling pathway of NF-κB and the expression of Bax and Bid in renal collecting duct epithelial cells of polycystic kidney disease (PKD), and to observe the tubulogenesis of TRAF3 transgenics for exploring the protective effect of TRAF3 on the cystogenesis and development of PKD. METHODS: The signaling changes of NF-κB and expression of Bax and Bid in PKD renal collecting duct epithelial cells and TRAF3 transgenic PKD renal collecting duct epithelial cells were determined by the Western blot. The percentage of apoptotic cells was measured by flow cytometry with Annexin V staining. The caspase-3 activity was also measured. The 3D Matrigel culture was performed to examine abnormal tubulomorphogenesis in vitro. RESULTS: The over-expression of TRAF3 significantly inhibited the signaling pathway of NF-κB in PKD renal collecting duct epithelial cells and significantly downregulated the expression of Bax and Bid, and also decreased the cell apoptosis. More branch structures were observed in the TRAF3 transgenic PKD renal collecting duct epithelial cells. CONCLUSION: TRAF3 is a negative regulatory inhibitor for Bax and Bid by regulating the NF-κB signaling and may be a new target for inhibiting the cyst formation in PKD.     

Effect of L-Arg on plasma content of endothelin and expression of c-fos mRNA in the left ventricle of rats with renovascular hypertensive hypertrophy

AIM:To study the effect of L-Arg on plasma content of endothelin (ET) and the expression of proto-oncogene c-fos mRNA in the left ventricle of rats with renovascular hypertensive hypertrophy. METHODS: The level of c-fos mRNA were measured by in situ hybridization. The ET in plasma were measured by radioimmunoassay. RESULTS:After eight weeks of treatment with L-Arg, the expression of c-fos decreased markedly (P＜0.01). The ET content in plasma also decreased significantly by L-Arg(P＜0.01).CONCLUSION: Plasma ET content and the expression of c-fos in the left ventricle of rats with renovascular hypertensive hypertrophy could be decreased by L-Arg administration.