Notch1 promotes renal tubulointerstitial fibrosis in renal tissues of diabetic mice by inhibiting autophagy through Akt/mTOR signaling pathway

AIM: To observe the changes of Notch1 expression and autophagy in the renal tissues of diabetic mice, and to explore the regulatory effect of Notch1 on tubulointerstitial fibrosis by inhibiting autophagy in diabetic nephro-pathy. METHODS: The mice were randomly divided into normal control group (db/m mice) and diabetes group (db/db mice), with 8 rats in each group. After 12 weeks of feeding, the mice were sacrificed and the corresponding biochemical indexes were measured. The protein expression of Notch1 in the renal tubular epithelial cells was observed by immunohistochemical staining. The protein levels of Notch1, PTEN, p-Akt (Thr308), Akt, p-mTOR (Ser2448), mTOR, LC3, P62, collagen type Ⅰ (Col-Ⅰ) and collagen type Ⅲ (Col-Ⅲ) were determined by Western blot. RESULTS: Compared with the db/m mice, the blood glucose, glycosylated hemoglobin, serum creatinine, triglyceride and total cholesterol were increased in the db/db mice (P<0.01). Renal tubular epithelial cell vacuolar degeneration, renal tubular expansion and interstitial inflammatory cell infiltration in db/db mouse renal tissues with HE staining were observed. The images of Masson staining showed collagenous fiber-like substance deposition in the glomerular capillaries and renal interstitium, and disarrangement of tubular structure in the renal tissues of db/db mice. The protein expression levels of PTEN and LC3-Ⅱ were decreased (P<0.01 or P<0.05), while the protein levels of Notch1, P62, p-mTOR (Ser2448), p-Akt (Thr308), Col-I and Col-III were increased in the db/db mice as compared with the db/m mice (P<0.01). However, no significant change of total mTOR and Akt proteins between the 2 groups was found. CONCLUSION: Notch1 protein expression was increased, PTEN expression was significantly reduced, Akt/mTOR pathway was activated, autophagy was inhibited, and fibrosis was aggravated in the renal tissues of the diabetic mice.     

Effect of HMGA2 gene on high glucose-induced apoptosis of renal tubular epithelial cells by regulating Notch signaling pathway

AIM:To investigate the effect of HMGA2 down-regulation on apoptosis and Notch signaling pathway in renal tubular epithelial cells exposed to high glucose (HG). METHODS:D-glucose at 5, 10, 20 and 30 mmol/L was used to stimulate human renal tubular epithelial HK-2 cells for 2 h, and D-glucose at 30 mmol/L was used to stimulate the HK-2 cells for 10 min, 60 min and 120 min. The protein expression of HMGA2 was determined by Western blot. The HK-2 cells were divided into normal glucose (NG) group, HG group, HG+si-HMGA2 group and HG+NC group, in which siRNA was transfected by Lipofectamine^TM 2000 for 48 h. Flow cytometry was used to analyze the apoptotic rate, reactive oxygen species (ROS) assay kit was used to detect ROS content, and Western blot was used to detect the protein levels of Notch1, Hes1 and Bcl-2. The HK-2 cells were treated with the Notch signaling pathway inhibitor DAPT, and then the cells were divided into HG group, HG+DAPT group and HG+si-HMGA2+DAPT group. The apoptotic rate was analyzed by flow cytometry. RESULTS:Exposure of the HK-2 cells to D-glucose at different concentrations for different time significantly increased the expression of HMGA2 (P<0.05). Compared with NG group, the protein expression of HMGA2, Notch1 and Hes1 in HG group was increased, the expression of Bcl-2/Bax was decreased, the apoptotic rate was increased, and the content of ROS was increased obviously (P<0.05). Compared with HG group, the protein expression of HMGA2, Notch1 and Hes1 of HG+si-HMGA2 group was decreased, the expression of Bcl-2/Bax was increased, the apoptotic rate was decreased, and the content of ROS was decreased significantly (P<0.05). The apoptotic rate in HG+DAPT group was significantly lower than that in HG group, while the apoptotic rate in HG+si-HMGA2+DAPT group was significantly lower than that in HG+DAPT group (P<0.05). CONCLUSION:Down-regulation of HMGA2 expression inhibits the apoptosis of renal tubular epithelial cells by regulating Notch signaling pathway and decreasing ROS production.     

Study on the Hepatoprotective Effect of Oxalis coriniculata L. and Related Mechanism by Regulating Oxidative Stress and TLR-2/NF-κB Signaling Pathway

[Objectives]This study aimed to explore the protective effect of Oxalis coriniculata L.on rats with acute liver injury induced by carbon tetrachloride(CCl4)and ...     

外源水杨酸诱导的部分甜瓜自毒作用相关基因的表达分析

以甜瓜(Cucumis melo)品种‘新银辉’为材料,采取根灌法,以水杨酸(10μmol/L)处理甜瓜幼苗,通过Real-Time PCR对叶片中部分自毒作用相关基因的表达量进行定量分析,以探究外源水杨酸诱导对甜瓜自毒作用的影响。结果显示,经外源水杨酸诱导后:(1)在催化苯丙烷类代谢途径的3个初始反应的蛋白酶编码基因中,PAL和C4H随处理后时间增加,呈逐渐上调趋势,但PAL比C4H更快响应胁迫,而4CL转录水平没有显著变化;(2)在类黄酮生物合成的3个关键酶的基因中,CHI、F3H均显示出不同程度的下调,CHS于处理后1 d略微上调,然后恢复至处理前水平;(3)两个转录因子基因WD40、R2R3-MYB分别在不同处理时间后显著下调;(4)WD40与CHI的表达相关性较高。据上述结果 ,水杨酸诱导后:(Ⅰ)CHI表达可能与转录因子WD40蛋白相关;(Ⅱ)WD40、R2R3-MYB与PAL、C4H、4CL、CHS间不存在简单、明确的调控关系;(Ⅲ)柚皮苷、表儿茶素、芦丁等甜瓜主要化感物质合成减少。     

烟酰胺腺嘌呤二核苷酸(NAD)合成途径和水稻叶片早衰

叶片是植物进行光合作用的主要场所,其衰老由内源遗传发育信号和外界环境胁迫所启动,是一个非常复杂有序的调控过程。烟酰胺腺嘌呤二核苷酸(nicotinamide adenine dinucleotide,NAD)是脱氢酶的辅酶,在糖酵解、糖异生、三羧酸循环以及呼吸链等代谢中发挥着不可替代的作用。最新研究表明,水稻NAD生物合成参与调控沉默信息调控因子Sirtuins的生物活性、组蛋白H3K9去乙酰化、植物激素茉莉酸(JA)和叶片衰老。本文综述了有关水稻叶片衰老的细胞生理特征、Sirtuins酶活、NAD生物合成以及水稻早衰的OsSRT1-NAD调控途径和OsSRT1-Me OH-JA调控途径,以期阐明水稻叶片衰老的分子机理及其调控途径,为高产育种提供相应的理论参考。     

Mechanism of JAK2-STAT3 signaling pathway in ischemic postconditio-ning neuroprotection after cerebral ischemia in tree shrews

AIM: To investigate the regulatory effect of JAK2-STAT3 signaling pathway on the neuroprotection of ischemic postconditioning (IPoC) in tree shrews, and to explore the mechanisms of cerebral injury deterioration after inhibiting the JAK2-STAT3 pathway. METHODS: The model of thrombotic cerebral ischemia was induced by photochemical reaction in tree shrews and the IPoC was established at 4 h after ischemia followed by clipping ipsilateral common carotid artery on the ischemia side for 5 min (3 times). After IPoC and intracerebroventricular injection of AG490 (JAK2 inhibitor), the changes of cerebral infarction area were detected by TTC staining, and the histological and ultrastructural changes of cortical neurons were observed under light and electron microscopes, respectively. The protein levels of t-STAT3 and p-STAT3 in the cortical tissue were determined by Western blot. RESULTS: The neuronal pycnosis, mitochondrial swelling and vanish of the mitochondrial cristae were found in cortical cortex, and the infarction area was (24.78±3.30)% at 24 h after cerebral ischemia. Meanwhile, the phosphorylation level of STAT3 protein in the cortical tissue was significantly increased (P<0.01). The cortical neuronal damage and mitochondrial swelling were decreased after IPoC, the area of cerebral infarction was significantly reduced to (17.67±1.83)% (P<0.01), and the phosphorylation level of STAT3 protein was further increased (P<0.01). However, the neuronal damage was aggravated, the infarction area was expanded to (23.85±2.77)%(P<0.05) after treatment with AG490, and the phosphorylation level of STAT3 protein was also significantly reduced (P<0.05). CONCLUSION: IPoC may reduce cerebral injury by regulating the phosphorylation of STAT3 protein, and inhibition of JAK2-STAT3 signaling pathway may counteract the cerebral protective effect of IPoC and aggravate brain injury.     

基于REE示踪的土壤团聚体破碎转化路径定量表征

土壤团聚体破碎转化路径是坡面侵蚀过程研究的难点问题之一。目前团聚体破碎转化路径的定量表征仍不明晰,一定程度上限制了土壤侵蚀过程中泥沙分选搬运机制的深入研究。基于大样带调查选取6种不同质地的典型农耕地土壤为研究对象,结合稀土元素(Rare Earth Elements,REE)示踪方法,综合分析不同粒径土壤团聚体(5~2,2~1,1~0.5,0.5~0.25,<0.25 mm)和不同径流扰动周期(24 h,7天)对REE吸附和解吸能力的影响,探究REE示踪不同粒径土壤团聚体破碎转化的可行性,定量表征了土壤团聚体破碎转化路径。结果表明:REE与土壤团聚体的实际吸附浓度低于施放浓度,2~1,1~0.5,0.5~0.25,<0.25 mm土壤团聚体的REE吸附浓度与黏粒含量呈显著正相关(P<0.05);径流扰动影响对吸附于土壤团聚体的REE解吸作用十分微弱,解吸浓度仅占REE实际吸附浓度的0.001%~0.139%。5~2,2~1,1~0.5,05~0.25,<0.25 mm土壤团聚体经过湿筛后向各粒径转化的路径基本相同,向<0.25 mm微团聚体转化为土壤团聚体破碎的主要路径。相较于粉粒、黏粒含量较高的土壤团聚体,砂粒含量较高的土壤团聚体向1~0.5,0.5~0.25 mm粒径的转化贡献率整体偏低。基于REE示踪得到的>0.25 mm各粒径团聚体质量整体被低估,低估范围为-27.96%^-11.08%;而<0.25 mm团聚体质量则被高估,高估范围为3.65%~22.73%。基于各粒径土壤团聚体的REE量化值建立了校正关系,可将计算相对误差降低至0.04%~16.24%。     

黄芩苷通过抑制核因子-κB和激活核因子-E2相关因子2信号通路缓解脂多糖诱导的小鼠空肠炎症和氧化应激

本试验旨在探讨黄芩苷对脂多糖(LPS)诱导的小鼠空肠炎症和氧化应激的缓解作用及其分子机制。将36只小鼠随机分为对照组、阴性对照组、模型组以及低、中、高剂量黄芩苷组,每组6只。适应性生长7 d后开始试验。对照组小鼠不作处理,阴性对照组和模型组小鼠每天灌胃0.2 mL磷酸盐缓冲液(PBS),黄芩苷组小鼠每天灌胃黄芩苷药液0.2 mL(根据小鼠体重计算黄芩苷用量,低剂量组为100 mg/kg BW,中剂量组为200 mg/kg BW,高剂量组为400 mg/kg BW),连续灌胃7 d,第7天给模型组、黄芩苷组小鼠腹腔注射0.2 mL LPS(3.5 mg/kg BW)。注射LPS 24 h后处死小鼠,取空肠进行形态学观察,检测空肠中炎性因子mRNA表达量和含量以及核因子-κB(NF-κB)和核因子-E2相关因子2(Nrf2)/血红素氧合酶-1(HO-1)信号通路关键蛋白表达量的变化。结果显示：相比于对照组,模型组小鼠空肠组织绒毛结构损伤,绒毛高度与隐窝深度比值显著降低(P<0.05),空肠中Toll样受体4(TLR4)的mRNA表达量显著升高(P<0.05),炎性因子[白细胞介...     

秸秆沼气工程化对环境的影响及应对措施

秸秆沼气工程产生的沼液在储存和使用过程中会对周围环境构成污染,其主要表现为沼液储存过程中有害气体释放、沼液直排造成的水体污染、农田长期大量施用造成的重金属沉积和沼液渗滤造成的地下水恶化。完善秸秆沼气工程储液池结构减少有害气体排放、建设配套沼液后处理设施、科学施用沼肥、开发沼液增值产品是解决秸秆沼气工程环境危害和促使其健康发展的重要技术途径。     

MC1R c.676A>G与山羊皮肤色素沉积的关系及其对黑色素合成的影响

为探讨山羊MC1R基因c.676A>G突变与山羊皮肤色素沉积的关系,及其对黑色素合成和cAMP信号通路下游色素相关基因表达的影响：一方面采用直接测序法检测酉州乌羊及其杂交后代群体(120个个体)MC1R基因c.676A>G的突变情况,用色差仪测量不同基因型个体的腹部皮肤色度值,分析c.676A>G突变与皮肤色素沉积的关系;另一方面,构建野生型(MC1R c.676A)和突变型(MC1R c.676G)真核表达载体,采用脂质体介导法转染到小鼠皮肤黑色素瘤细胞(B16-F10)中进行过表达,利用酶标仪检测各组细胞黑色素含量的差异,并通过实时荧光定量PCR方法检测各组细胞中cAMP信号通路下游色素相关基因(MITF、TYR、TYRP1、DCT)的表达差异性。结果显示：酉州乌羊及其杂交后代群体中,c.676A>G位点均以GG基因型占优势,不同基因型群体的皮肤色度值差异不显著(P>0.05)。MC1R突变组(MC1R c.676G)细胞中的黑色素含量极显著(P<0.05)高于MC1R野生组(MC1R c.676A)。MITF、TYRP1和DCT基因在MC1R突...