草莓CIPK基因家族的鉴定与表达分析

从拟南芥数据库中获得CIPK基因家族注册号，运用生物信息学分析的方法，在蔷薇科森林草莓（Fragaria vesca）数据库中得到CIPK基因家族成员19个，可分为6个亚族。该基因家族分布在草莓7条染色体中的6条上。其编码蛋白的氨基酸数157 ~ 1 196，理论等电点3.91 ~ 9.34，分子量18 667.68 ~ 133 714.31 D。基因结构分析表明，有11条基因只有1个外显子，其余基因外显子数2 ~ 15。亚细胞定位结果表明，该基因家族成员主要在细胞质、细胞核和叶绿体上表达。蛋白质二级结构预测表明，该基因家族成员主要以α–螺旋、β–转角和不规则卷曲为主。对上游2 kb区域启动子顺式作用元件分析表明，该基因家族成员对逆境胁迫应答MYB响应明显，除FvCIPK02、FvCIPK15、FvCIPK17外，其他基因均对脱落酸应答元件ABRE响应明显。qRT-PCR数据分析表明，FvCIPK16、FvCIPK10和FvCIPK09分别在PEG、ABA和NaCl处理下，草莓试管苗中的相对表达量最高，分别是对照的18.4倍、29倍、13倍，说明FvCIPK16强响应干旱胁迫，FvCIPK10强响应ABA诱导，FvCIPK09强响应高盐胁迫；另外发现各处理的FvCIPK03相对表达量均下调，推测FvCIPK03在植物逆境胁迫中起负调节作用。

Effects of alpha-lipoic acid on MPTP-induced Parkinson disease in mice

AIM: To investigate the neuroprotective effect of alpha-lipoic acid (ALA) in mice with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced Parkinson disease (PD). METHODS: After establishment of the MPTP-induced PD mouse model, the effects of ALA on the motor deficits in PD mice were evaluated by open-field test, hanging wire test and rotarod test. The effects of ALA on dopaminergic neurons in substantia nigra and striatum of the PD mice were detected by immunohistochemical staining. The effects of ALA on the release of dopamine (DA) and its metabolite 3,4-dihydroxyphenylacetic acid (DOPAC) in striatum of PD mice were measured by high-performance liquid chromatography. The effects of ALA on activation of microglia in PD mice and the expression of inducible nitric oxide synthase (iNOS) in microglia were examined by immunofluorescence staining. The effects of ALA on the expression of tyrosine hydroxylase (TH), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and cyclooxygenase-2 (COX-2) in striatum of PD mice were determined by Western blot. RESULTS: ALA treatment significantly increased the total moving distance, average velocity and hanging wire test score (P < 0.05), but significantly decreased the falling lantency of rotarod test (P < 0.01) ALA treatment significantly increased the expression levels of TH in substantia nigra and striatum (P < 0.01), and the release levels of DA and DOPAC in striatum (P < 0.01). ALA treatment significantly decreased microglia activation and iNOS positive microglia in substantia nigra (P < 0.01). ALA treatment significantly increased the expression level of TH, significantly decreased the expression levels of IL-1β, TNF-α and COX-2 in the striatum (P < 0.01). CONCLUSION: ALA plays a neuroprotective role in the mice with MPTP-induced PD. ALA reduces microglial cell activation, dopaminergic neuron damage and neuroinflammation, and attenuates motor deficits in MPTP-induced PD mice.