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.     

Panax notoginseng saponins suppress oxygen-glucose deprivation/reoxygenation-induced pyroptosis of SH-SY5Y cells

AIM To investigate the effect of Panax notoginseng saponins (PNS) on pyroptosis of SH-SY5Y cells induced by oxygen-glucose deprivation/reoxygenation (OGD/R). METHODS The OGD/R was conducted to induce ischemia/reperfusion injury in SH-SY5Y cells. The effects of PNS on the viability (detected by CCK-8 assay) and membrane permeability [indicated by lactate dehydrogenase (LDH) leakage and propidium iodide (PI) staining positive cell proportion] of OGD/R-induced SH-SY5Y cells were observed. The protein levels of gasdermin D (GSDMD), GSDMD N-terminal fragment (GSDMD-N), caspase-1 and caspase-4, and the release of interleukin-1β (IL-1β) and IL-18 in the cells were also determined. RESULTS After exposure to OGD/R, the viability of SH-SY5Y cells dramatically decreased (P<0.01), while the LDH leakage, the PI staining positive cell proportion, the protein levels of GSDMD, GSDMD-N, caspase-1 and caspase-4, and the release of IL-1β and IL-18 were significantly increased (P<0.01). However, PNS treatment enhanced the viability of SH-SY5Y cells inhibited by OGD/R (P<0.01), but reduced the leakage of LDH and the percentage of PI staining positive cells (P<0.05 or P<0.01). Moreover, PNS reversed the increases in the protein levels of GSDMD, GSDMD-N, caspase-1 and caspase-4 and the release of IL-1β and IL-18 in OGD/R-induced SH-SY5Y cells (P<0.05 or P<0.01). CONCLUSION Treatment with PNS alleviates OGD/R-induced injury in SH-SY5Y cells. Its mechanism may be related to inhibition of SH-SY5Y cell pyroptosis induced by OGD/R.     

基于电项针对失眠大鼠下丘脑免疫细胞因子影响的实验研究

目的 观察不同频率电项针对睡眠剥夺大鼠下丘脑内免疫因子白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）含量的影响，明确电项针对失眠的治疗作用，并探讨其发挥促眠效应的可能作用机制。方法 将50只健康雄性Wistar大鼠随机分成空白组、模型组、2 Hz刺激组、50 Hz刺激组、100 Hz刺激组，每组10只。采用对氯苯丙氨酸（PCPA）化法制作失眠大鼠模型。造模成功后，各频率刺激组连续1周进行电项针干预治疗，1次/日，30 min/次，空白组和模型组每日在相应时间仿照刺激组固定躯体，不进行任何处置。各组于治疗结束后处死大鼠后迅速分离下丘脑组织，运用酶联免疫吸附法（ELISA）测定IL-1β、IL-6和TNF-α含量。结果 模型组大鼠下丘脑中IL-1β、IL-6和TNF-α浓度明显低于空白组，差异具有统计学意义（P<0.05）。与模型组比较，各频率刺激组IL-1β、IL-6和TNF-α浓度均升高，差异具有统计学意义（P<0.05）；各频率刺激组之间比较，2 Hz刺激组下丘脑IL-1β、IL-6和TNF-α含量高于50 Hz和100 Hz组（P<0.05），50Hz组和100Hz组间比较无统计学差异（P>0.05）。结论 不同频率电项针治疗失眠均有效，各频率组电项针治疗比较，2Hz刺激组的促眠作用最好，其促眠作用的机制可能与调节下丘脑内免疫因子IL-1β、IL-6和TNF-α的含量相关。     

适度饥饿浅黄恩蚜小蜂对烟粉虱和温室白粉虱的寄生和取食选择

为评价释放前经历饥饿对浅黄恩蚜小蜂Encarsia sophia(GiraultDodd)寄生取食粉虱能力的影响,以3龄Q隐种烟粉虱Bemisia tabaci Q和温室白粉虱Trialeurodes vaporariorum若虫为寄主,在2种粉虱单独或同时存在的情况下,比较释放前经适度饥饿、初羽化未饥饿和初羽化喂饲蜂蜜水3种处理的浅黄恩蚜小蜂对2种粉虱寄主的寄生和取食选择情况。结果表明,在2种粉虱单独存在时,经适度饥饿6 h的浅黄恩蚜小蜂寄生的烟粉虱和温室白粉虱显著多于其它处理,而且能取食更多的温室白粉虱,经适度饥饿的寄生蜂在24 h内通过寄生和取食杀死烟粉虱和温室白粉虱的总量分别为12.5头和12.9头。在2种粉虱同时存在时,适度饥饿寄生蜂取食2种粉虱的总量明显高于其它处理,但不同处理间无显著差异,适度饥饿寄生蜂通过寄生和取食杀死2种粉虱的数量最多为11.5头,显著高于未饥饿处理的6.5头。表明释放前经历适度饥饿可以明显提高浅黄恩蚜小蜂寄生和取食粉虱若虫的能力。     

Improved porcine model for Shiga toxin‐producing Escherichia coli infection by deprivation of colostrum feeding in newborn piglets

Porcine edema disease (ED) is a toxemia caused by enteric infection with Shiga toxin 2e (Stx2e)‐producing Escherichia coli (STEC). ED occurs most frequently during the weaning period and is manifested as emaciation associated with high mortality. In our experimental infection with a specific STEC strain, we failed to cause the suppression of weight gain in piglets, which is a typical symptom of ED, in two consecutive experiments. Therefore, we examined the effects of deprivation of colostrum on the sensitivity of newborn piglets to STEC infection. Neonatal pigs were categorized into two groups: one fed artificial milk instead of colostrum in the first 24 h after birth and then returned to the care of their mother, the other breastfed by a surrogate mother until weaning. The oral challenge with 10¹¹ colony‐forming units of virulent STEC strain on days 25, 26 and 27 caused suppression of weight gain and other ED symptoms in both groups, suggesting that colostrum deprivation from piglets was effective in enhancing susceptibility to STEC. Two successive STEC infection experiments using colostrum‐deprived piglets reproduced this result, leading us to conclude that this improved ED piglet model is more sensitive to STEC infection than the previously established models.     

小麦小分子RNA TaMIR1129介导植株抵御低氮逆境功能研究

本研究针对迄今有关小麦小分子RNA(miRNA)家族成员介导植株氮素吸收和利用机理尚少见报道的现状,对TaMIR1129的表达特征和介导植株抵御低氮逆境功能进行了研究。结果表明,TaMIR1129呈低氮胁迫诱导表达,表现为随氮浓度降低(0.02~6mmol/L)和处理时间延长(0~48h)表达水平不断增高特征。此外,低氮诱导的高表达水平在恢复供氮后表达下调。表明该miRNA对介质中氮素应答呈典型的时间及浓度依赖特征。TaMIR1129作用2个靶基因,包括Molybdenum cofactor sulfurase(TaMCS)和Major facilitator family transporter(TaMFFT),上述基因应答低氮特征与TaMIR1129相反。遗传转化结果表明,超表达TaMIR1129具有显著增强植株抵御低氮逆境的能力。表现为与野生型对照相比,转基因系Sen 1和Sen 2低氮处理后植株形态增大,干质量增加,氮累积量增多。表明TaMIR1129与作用靶基因构建miRNA/target模块在介导植株抵御低氮逆境中发挥重要作用。     

Protective effect of progesterone on PC12 cells injured by oxygen-glucose deprivation

AIM: To investigate the effects of progesterone on the cell viability and expression of glucose transporter type 3(GLUT3) in PC12 cells injured by oxygen-glucose deprivation (OGD) in attempt to prove the neuroprotection of progesterone (PROG) against the hypoxic-ischemic injury in cultured cells in vitro. METHODS: Well-differentiated PC12 cells induced by nerve growth factor were randomly divided into 3 groups. In normal group, the cells were cultured without OGD treatment. In OGD group, the culture medium was replaced by glucose-free medium and the cells were transferred to a humidified incubation chamber flushed by a gas mixture of 95% N₂ and 5% CO₂ for 30 min. After that, the cells were fed with glucose-supplemented medium and cultured under normoxic condition for 24 h. In PROG+OGD group, the cells were given the same treatments as those in OGD group except that the medium contained progesterone at concentration of 10 nmol/L. Cellular morphological changes were observed after OGD for 30 min. The cell viability was assessed by WST-8 assay. The degree of the cell damage was evaluated by determining lactate dehydrogenase (LDH) leakage. The expression of GLUT3 at mRNA and protein levels was examined by RT-PCR and Western blotting, respectively. RESULTS: Progesterone attenuated the cellular swelling, decreased the leakage of LDH and improved the viability of PC12 cells injured by OGD (P<0.01). The expression of GLUT3 at mRNA and protein levels in PC12 cells in PROG+OGD group was significantly higher than that in OGD group (P<0.05). CONCLUSION: Progesterone has protective effect on in vitro cultured PC12 cells injured by OGD. The mechanism may be related to the up-regulation of GLUT3 protein.     

持续气道正压通气治疗阻塞型睡眠呼吸暂停低通气综合征合并2型糖尿病的疗效观察

目的通过观察持续气道正压通气（CPAP）治疗对合并2型糖尿病（T2DM）的阻塞型睡眠呼吸暂停低通气综合征（OSAHS）患者的胰岛素抵抗（IR）的影响。方法 46例合并T2DM的OSAHS患者随机分为2组,对照组（22例）行保守治疗,而实验组（24例）加用CPAP治疗。观察所有患者治疗前及治疗后1、3、6个月的临床指标变化,采用稳态模式评估（HOMA）指数评价IR。结果实验组患者治疗1个月后,其最低SaO 2水平明显高于治疗前及同一时段的对照组（P〈0.01）;治疗3个月后,其呼吸暂停低通气指数明显低于治疗前及同一时段的对照组（P〈0.01）;治疗6个月后,其空腹胰岛素及HOMA水平明显低于治疗前及同一时段的对照组（P〈0.01）。结论长期CPAP治疗可显著改善合并T2DM的OSAHS患者的IR。