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Effects of lanthanide ions on hydrolysis of phosphatidylin-ositol in human erythrocyte membranes

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Chinese Science Bulletin 2001年第5期46卷 401-403页

作者： huixue liu Jian Hu Xiangtao liu Rongchang Li Kui Wang Beijing Med Univ Sch Pharmaceut Sci Dept Biol Chem Beijing 100083 Peoples R China

The effects of lanthanides on the hydrolysis of phosphatidylinositol in human erythrocyte membranes were studied. 3H-inositoI labeling chromatography and HPLC were used to determine inositol 1, 4, 5-triphosphatc and dia-cylglycerol separately, the hydrolytic products of phosphatidylinositol due to the reaction of lanthanide ions with human erythrocyte membranes. The unhydrolyzed phosphatidylinositol in membranes was also determined. The results indicate that the hydrolysis of phosphatidylinositol can be promoted by lanthanides (La3+, Ce3+, Y3+, Tb3+) and the effects of La3+ and Ce3+ are stronger than those of Y3+ and Tb3+.

关键词： lanthanids ions phosphatidylinositol second messengers.

Vanadyl ions binding to GroEL (HSP60) and inducing its depolymerization

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Chinese Science Bulletin 2007年第20期52卷 2775-2781页

作者： LEI WanHua liu huixue ZHONG LiJun YANG XiaoDa WANG Kui Department of Chemical Biology School of Pharmaceutical Sciences Peking University Health Science Center Beijing 100083 China Medical and Healthy Analysis Center Peking University Health Science Center Beijing 100083 China National Laboratory of Natural and Biomimetric Drugs School of Pharmaceutical Sciences Peking University Health Science Center Beijing 100083 China

Several vanadium compounds have been known for the hypoglycemic and anticancer effects. However, the mechanisms of the pharmacological and toxicological effects were not clear. In this work, we in- vestigated the potential targets of vanadium in mitochondria. Vanadyl ions were found to bind to mi- tochondria from rat liver with a stoichiometry of 244±58 nmol/mg protein and an apparent dissocia- tion constant (Kd) of (2.0±0.8)×10·16 mol/L. Using size exclusion chromatography, a vanadium-binding protein was isolated and identified to be the 60-kDa heat shock protein (HSP60) by mass spectrometry analysis and immunoassays. Additionally, binding of vanadyl ions was found to result in depolymeri- zation of homo-oligomeric HSP60 (GroEL). HSP60 is an indispensable molecular chaperone and in- volved in many kinds of pathogenesis of inflammatory and autoimmune diseases, e.g. type 1 diabetes. Our results suggested that HSP60 could be a novel important target involved in the biological and/or toxicological effects of vanadium compounds.

关键词：糖尿病线粒体钒疗效

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细胞线粒体状态的热力学分析

化学进展 2006年第6期18卷 841-848页

作者：刘莎刘会雪杨晓改杨晓达北京大学药学院化学生物学系北京100083

近年来,线粒体在一些重大的代谢性疾病如糖尿病和神经退行性病变中的作用备受关注。由于线粒体是细胞生产能量的“工厂”,线粒体的能量代谢理论研究带动了线粒体热力学研究的进展,也向化学热力学研究提出新的问题。本文在总结了化学渗... 详细信息

近年来,线粒体在一些重大的代谢性疾病如糖尿病和神经退行性病变中的作用备受关注。由于线粒体是细胞生产能量的“工厂”,线粒体的能量代谢理论研究带动了线粒体热力学研究的进展,也向化学热力学研究提出新的问题。本文在总结了化学渗透理论和近年来与线粒体有关的生物化学研究新结果的基础上,对线粒体的工作及其调节机制进行了分析,提出了线粒体工作状态受化学热力学机制控制的新观点。

关键词：线粒体化学渗透理论质子动势离子通道化学热力学

Amavadin induced PTP opening not through the promotion of ROS generation in rat kidney mitochondria

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Journal of Chinese Pharmaceutical Sciences 2014年第12期23卷 830-836页

作者：霍辰伊刘会雪北京大学医学部药学院化学生物学系北京100191

Amavadin is a natural vanadium compound that accumulates to high level in poisonous Amanita mushrooms. Recently, amavadin was found to have potential therapeutic effect in cancer treatment. However, its toxicity and the possible mechanism of actions are still not clear. In this study, we investigated the toxic effects of amavadin on rat kidney mitochondria and the possible mechanism. We found that amavadin induced significantly permeability transition pore （PTP） opening in the mitochondria. Amavadin y inhibited the generation of reactive oxygen species （ROS） in succinate buffer, and at high concentration of 200 gM it increased the ROS generation in malate buffer. With the addition of rotenone, the ROS generation in malate buffer was strongly enhanced than that induced by amavadin alone, but remained unchanged in succinate buffer. Results from the present study suggest that amavadin act upon electron transport chain downstream of rotenone, and the ubiquinone binding site in complex I is the most possible binding site.

关键词： Amavadin Vanadium compound Mitochondria Reactive oxygen species Permeability transition pore

Lanthanum chloride or citrate is absorbed mainly via M cells in gastrointestinal tracts with lanthanum phosphates as the transformed species

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Journal of Chinese Pharmaceutical Sciences 2018年第8期27卷 553-564页

作者： Huixia Huang huixue liu Xiaojie Ma Hui Guan Xiaogai Yang Department of Chemical Biology School of Pharmaceutical SciencesPeking University Health Science CenterBeijing 100191China

In the present study, we investigated the transformed species and the absorptive mechanism of rare earth elements（REEs） in gastrointestinal（GI） tract, using La Cl3 and La Cit as representative compounds. Artificial gastric and intestinal fluids were used to simulate the environment of the digestive tract in vivo. The inductively coupled plasma mass spectrometry（ICP-MS） result showed that more than 99.9% of La Cl3 and La Cit formed precipitation in artificial intestinal fluid, with the average size distribution of 200 nm（2-h incubation） increasing to 600 nm（24-h incubation） determined by dynamic light scattering（DLS）, indicating the aggregation of the particles. The Fourier transform infrared spectroscopy（FTIR） analysis demonstrated that the constituents of these particles were mainly in the form of lanthanum phosphates. To explore the transport mechanism of REEs in GI tract, the mice Peyer＇s patches（PPs） and intestinal epithelium were separated to evaluate the content of lanthanum by ICP-MS following oral administration with 2 or 100 mg/kg/day of La Cit for 7 d. The results showed that the amount of lanthanum phosphate particles absorbed by PPs was significantly greater than that of intestinal epithelium, indicating that lanthanum particles might be phagocytosed mainly by M cells located in the follicle-associated epithelium（FAE） overlying PPs. Furthermore, Caco-2 cell monoculture and Caco-2/Raji B cell coculture models were established to simulate intestinal epithelial cells and FAE, respectively. The result showed that the transport of lanthanum in Caco-2/Raji B coculture model was significantly higher than that in Caco-2 monoculture model（about 60 times higher）, and the level of lanthanum in the basal compartment of Caco-2 monoculture model was very low, supporting that M cells were the main route for lanthanum phosphate particles to be transported and absorbed. Taken together, these data suggested that La Cl3 and La Cit in GI tract were absorbed mainly via M cel

关键词： Lanthanum chloride (LaCl3) Lanthanum citrate (LaCit) Lanthanum phosphates Artificial intestinal fluid Peyer's patches M cells Caco-2 cell monoculture model Caco-2/Raji B cells coculture model Transport and absorption

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华北落叶松人工林林缘光照度研究

河北林果研究 2014年第4期29卷 356-358,363页

作者：刘会学雄县农业局河北雄县071800

林内光照环境是林分更新的基础。本试验以华北落叶松人工林为研究对象,采用30m的样带法,比较了林分内部及林缘光照度的变化规律。结果表明:华北落叶松人工林林缘相对光照度随距林缘距离的增加而减小,之后趋于稳定;密度为630株/hm2时,随... 详细信息

林内光照环境是林分更新的基础。本试验以华北落叶松人工林为研究对象,采用30m的样带法,比较了林分内部及林缘光照度的变化规律。结果表明:华北落叶松人工林林缘相对光照度随距林缘距离的增加而减小,之后趋于稳定;密度为630株/hm2时,随林缘距离的变化不太明显;林缘相对光照度与林分边缘不同距离呈明显的二次多项式关系;相对光照度能较好地反映林缘效应;随林分密度的增加林内光照度逐渐降低,相对光照度与密度呈显著幂函数关系。

关键词：华北落叶松林缘光照度

不同水合粒径的磷酸镧微粒对巨噬细胞IL-1β分泌水平的影响以及与其入胞途径的相关性

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中国稀土学报 2023年第3期41卷 631-640,I0006页

作者：孙冀芳刘会雪杨晓达杨晓改王夔北京大学药学院北京100191

基于稀土的应用现状,本研究以人源THP-1巨噬细胞(THP-M)为研究模型,选取了具不同水合粒径大小的LaPO_4微粒,研究了它们刺激THP-M IL-1β分泌水平的差异性以及入胞途径与IL-1β分泌水平的相关性。研究表明,LaCl_3在含不同血清的培养基中... 详细信息

基于稀土的应用现状,本研究以人源THP-1巨噬细胞(THP-M)为研究模型,选取了具不同水合粒径大小的LaPO_4微粒,研究了它们刺激THP-M IL-1β分泌水平的差异性以及入胞途径与IL-1β分泌水平的相关性。研究表明,LaCl_3在含不同血清的培养基中可形成水合粒径大小不同的磷酸镧微粒;水合粒径越大,刺激THP-M分泌IL-1β的水平也越强;水合粒径约为1μm的磷酸镧微粒主要通过吞噬作用和巨胞饮以及网格蛋白介导的内吞进入细胞,对其介导的IL-1β分泌贡献较大;但陷窝蛋白介导的内吞途径对具较小水合粒径微粒的细胞摄入以及刺激细胞分泌IL-1β的水平也有较大贡献。研究表明具不同水合粒径大小的稀土微粒可通过不同的跨膜途径进入细胞,从而引起其刺激细胞分泌IL-1β的效应的差异性。这提示微粒的大小以及在体内的入胞方式与稀土微粒引起的炎症反应相关,将为稀土的作用模式以及在胞内分布和转运的方式提供依据,有助于稀土的风险性评估和合理应用。

关键词：稀土磷酸盐微粒巨噬细胞 IL-1β 内吞

三七皂甙R1调节蛋白酪氨酸激酶2/信号转导与转录激活因子3信号通路对肺炎克雷伯菌感染的肺泡上皮细胞损伤的影响

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中国热带医学 2023年第7期23卷 754-760页

作者：刘会雪张佩佩安江科石家庄平安医院呼吸科河北石家庄050000 石家庄平安医院重症医学科河北石家庄050000

目的探讨三七皂甙R1(notoginsenoside R1, NGR1)对肺炎克雷伯菌(Klebsiella pneumoniae, Kp)感染的肺泡上皮细胞损伤及蛋白酪氨酸激酶2/信号转导与转录激活因子3(janus kinase signal transducers 2 and activator of transcription 3, JAK2/STAT3)信号通路的影响。方法 A549细胞分为5组:对照组(C组)、感染组(Infect组)、感染+低NGR1组(Infect+L-NGR1组)、感染+高NGR1组(Infect+H-NGR1组)、感染+高NGR1+JAK2/STAT3通路抑制剂组(Infect+HNGR1+SD-1029组)。采用CCK-8测定细胞增殖;ELISA试剂盒检测培养液白细胞介素-1β(interleukin-1β, IL-1β)、肿瘤坏死因子-α(tumor necrosis factor-α, TNF-α)、γ干扰素(interferon γ, IFN-γ)含量;流式细胞术检测细胞凋亡;RT-qPCR检测JAK2、STAT3表达;Western Blot检测JAK2/STAT3通路、自噬蛋白微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3, LC3)、自噬相关基因5(autophagy-relatedgene5, Atg5)自噬关键分子酵母Atg6同系物(autophagyrelated gene 6, Beclin1)、凋亡蛋白B细胞淋巴瘤/白血病-2蛋白(B-cell lymphoma 2, bcl-2)、Bcl-2相关x蛋白(bcl-2 associated protein, Bax)、半胱氨酸蛋白酶3(cysteinyl aspartate specific proteinase, Caspases3)蛋白表达。结果与C组相比,Infect组72 h细胞活力、Bcl-2、LC3-Ⅱ/Ⅰ、Atg5、Beclin-1蛋白水平、JAK2、STAT3 mRNA相对表达量及蛋白磷酸化水平显著降低(P<0.05),IL-1β、TNF-α、IFN-γ含量、细胞凋亡率、Bax、cleaved-Caspase-3蛋白水平显著升高(P<0.05)。与Infect组相比,Infect+L-NGR1组、Infect+H-NGR1组72 h细胞活力、Bcl-2、LC3-Ⅱ/Ⅰ、Atg5、Beclin-1蛋白水平、JAK2、STAT3 mRNA相对表达量及蛋白磷酸化水平显著上升(P<0.05),IL-1β、TNF-α、IFN-γ含量、细胞凋亡率、Bax、cleaved-Caspase-3蛋白水平显著下降(P<0.05)。与Infect+H-NGR1组相比,Infect+H-NGR1+SD-1029组72 h细胞活力、Bcl-2、LC3-Ⅱ/Ⅰ、Atg5、Beclin-1蛋白水平、JAK2、STAT3蛋白磷酸化水平显著降低(P<0.05),IL-1β、TNF-α、IFN-γ含量、细胞凋亡率、Bax、cleaved-Caspase-3蛋白水平显著升高(P<0.05)。结论 NGR1可激活JAK2/STAT3信号通路,促进肺泡上皮细胞自噬,抑制Kp诱导的肺泡上皮细胞炎症损伤和凋亡。

关键词：三七皂甙R1 JAK2/STAT3信号通路肺炎克雷伯菌肺泡上皮细胞炎症自噬

稀土离子(La^(3+),Gd^(3+),Yb^(3+),Ce^(3+))对线粒体的氧化损伤

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中国稀土学报 2007年第1期25卷 102-106页

作者：刘会雪杨晓达王夔北京大学药学院化学生物学系北京100083

主要考察了稀土离子对线粒体氧化损伤(膜脂质过氧化、膜蛋白氧化、线粒体DNA氧化)的作用。结果表明,稀土离子(La3+,Gd3+,Yb3+,Ce3+)浓度大于2×10-5mol.L-1时,对线粒体膜脂质过氧化、膜蛋白氧化均有明显的促进作用;在对Fe2+诱导的线粒... 详细信息

主要考察了稀土离子对线粒体氧化损伤(膜脂质过氧化、膜蛋白氧化、线粒体DNA氧化)的作用。结果表明,稀土离子(La3+,Gd3+,Yb3+,Ce3+)浓度大于2×10-5mol.L-1时,对线粒体膜脂质过氧化、膜蛋白氧化均有明显的促进作用;在对Fe2+诱导的线粒体氧化过程中,La3+,Gd3+和Yb3+能明显地增强Fe2+的氧化作用,而Ce3+对Fe2+的作用表现出明显的拮抗作用,此外Ce3+对Fe2+诱导的线粒体DNA氧化损伤表现出明显的拮抗作用,显示了Ce3+的特殊性。

关键词：镧线粒体氧化性损伤稀土