Lovastatin protects mesenchymal stem cells against hypoxia- and serum deprivation-induced apoptosis by activation of PI3K/Akt and ERK1/2

Cell therapy with bone marrow-derived mesenchymal stem cells (MSCs) has been shown to have great promises in cardiac repair after myocardial infarction. However, poor viability of transplanted MSCs in the infracted heart has limited the therapeutic efficacy. Our previous studies have shown in vitro that rat MSCs undergo caspase-dependent apoptosis in response to hypoxia and serum deprivation (Hypoxia/SD). Recent findings have implicated statins, an established class of cholesterol-lowering drugs, enhance the survival of cells under various conditions. In this study, we investigated the effect of lovastatin on rat MSCs apoptosis induced by Hypoxia/SD, focusing in particular on regulation of mitochondrial apoptotic pathway and the survival signaling pathways. We demonstrated that lovastatin (0.01-1 microM) remarkably prevented MSCs from Hypoxia/SD-induced apoptosis through inhibition of the mitochondrial apoptotic pathway, leading to attenuation of caspase-3 activation. The loss of mitochondrial membrane potential and cytochrome-c release from mitochondria to cytosol were significantly inhibited by lovastatin. Furthermore, the antiapoptotic effect of lovastatin on mitochondrial apoptotic pathway was effectively abrogated by both PI3K inhibitor, LY294002 and ERK1/2 inhibitor, U0126. The phosphorylations of Akt/GSK3 beta and ERK1/2 stimulated by lovastatin were detected. The activation of ERK1/2 was inhibited by a PI3K inhibitor, LY294002, but U0126, a ERK1/2 inhibitor did not inhibit phosphorylation of Akt and GSK3 beta. These data demonstrate that lovastatin protects MSCs from Hypoxia/SD-induced apoptosis via PI3K/Akt and MEK/ERK1/2 pathways, suggesting that it may prove a useful therapeutic adjunct for transplanting MSCs into damaged heart after myocardial infarction.     

Specific LPA receptor subtype mediation of LPA-induced hypertrophy of cardiac myocytes and involvement of Akt and NFkappaB signal pathways

Lysophosphatidic acid (LPA) is a bioactive phospholipid with diverse functions mediated via G-protein-coupled receptors (GPCRs). In view of the elevated levels of LPA in acute myocardial infarction (MI) patients we have conducted studies aimed at identifying specific LPA receptor subtypes and signaling events that may mediate its actions in hypertrophic remodeling. Experiments were carried out in cultured neonatal rat cardiomyocytes (NRCMs) exposed to LPA and in a rat MI model. In NRCMs, LPA-induced hypertrophic growth was completely abrogated by DGPP, an LPA1/LPA3 antagonist. The LPA3 agonist OMPT, but not the LPA2 agonist dodecylphosphate, promoted hypertrophy as examined by 3[H]-Leucine incorporation, ANF-luciferase expression and cell area. In in vivo experiments, LPA1, LPA2 and LPA3 mRNA levels as well as LPA1 and LPA3 protein levels increased together with left ventricular remodeling (LVRM) after MI. In addition, LPA stimulated the phosphorylation of Akt and p65 protein and activated NF-kappaB-luciferase expression. Inhibitors of PI3K (wortmannin), mTOR (rapamycin), and NF-kappaB (PDTC or SN50) effectively prevented LPA-induced 3[H]-Leucine incorporation and ANF-luciferase expression. Furthermore, ERK inhibitors (U0126 and PD98059) suppressed LPA-stimulated activation of NF-kappaB and p65 phosphorylation whereas wortmannin showed no effect on NF-kappaB activation. Our findings indicate that LPA3 and/or LPA1 mediate LPA-induced hypertrophy of NRCMs and that LPA1 and LPA3 may be involved in LVRM of MI rats. Moreover, Akt and NF-kappaB signaling pathways independently implicate in LPA-stimulated myocardial hypertrophic growth.     

薄层层析-紫外分光光度法测定十全大补胶囊中芍药苷含量

目的 建立薄层层析-紫外分光光度法测定十全大补胶囊中芍药苷含量的方法。方法 使用硅胶GF254薄层板,醋酸乙酯：甲酸：冰醋酸：水(30：2：2：4)为展开剂,在紫外光灯(254nm)下定位,230nm波长处检测。结果 点样量在25～125μg范围内呈现良好的线性关系,其回归方程为Y=0．004658C-0．01959,r=0．9994,平均回收率为99．20％,RSD=1．19％。结论 本法准确,适用于该药的质量控制。     

膜蛋白结构研究方法新进展

膜蛋白是一类结构独特的蛋白质,执行很多基本的和重要的细胞生物学功能。了解膜蛋白在生物膜上的基本构象,对研究膜蛋白的精细拓扑结构、功能具有重要意义。但是膜蛋白的疏水特性使其需要与生物膜共同形成稳定的自然构象,至今在蛋白质组学的研究中对膜蛋白知之甚少。了解分子结构是了解生物大分子功能的一个重要途径。因此,本文对近来膜分子结构研究领域的进展作一简要概述。晶体学方法、单颗粒方法和原子力显微镜为膜蛋白的研究提供了大量的细节数据。固相核磁共振技术提供了跨膜α螺旋结构的方向约束数据和精确的分子间距离约束数据。直接位点标记的旋转电子顺磁共振可以得到更长的距离约束数据,但是目前的标记策略仍然具有局限性。位点特异的红外二色性分析可使得在脂双层中定向分析跨膜α螺旋束成为可能。     

PCR条件对扩增SARS-CoV多聚酶部分基因的影响

目的：对该实验室已建立的检测SARS冠状病毒多聚酶基因的套式RT-PCR方法进行优化。方法：从SAPS病人的嗽口水标本中提取RNA,调整套式PCR的退火温度,扩增SARS冠状病毒多聚酶部分基因。扩增出的阳性片段连接入pGEM-T载体中,测序后比较其与已知SARS冠状病毒的同源性。结果：通过改变PCR条件,成功从一SARS病人的嗽口水中扩增出SARS冠状病毒多聚酶部分基因。结论：针对不同标本优化PCR反应条件非常重要。     

杨树根系浸提液对3种牧草种子萌发及幼苗生长的影响

采用不同浓度(3、10和30mg/mL)的南林95杨〔PopulusdeltoidsBartr.cv.Lux×P.euramericana(Dode)Guineircv.I 45/51〕和NL 80351杨(P.deltoidsBartr.cv.Lux×P.deltoidsBartr.cv.Harvard)根系浸提液,处理杨树 牧草复合经营系统中3种主要牧草种子,探讨杨树根系浸提液对牧草种子萌发及幼苗生长的影响。结果表明:南林95杨和NL 80351杨根系浸提液对3种牧草种子的萌发及幼苗的生长有一定影响。这2个无性系根系浸提液对紫花苜蓿(MedicagosativaL.)、白三叶(TrifoliumrepensL.)和杂交狼尾草(PennisetumalopecuroidesL.×P.americanumL.)的种子发芽率、幼苗高生长、根长生长及种子活力主要表现为抑制作用,在浓度较高时影响尤为明显;但2个无性系根系浸提液对白三叶的高生长(南林95杨30mg/mL除外)表现出一定的促进作用;在3mg/mL浓度时,2个无性系根系浸提液对白三叶的种子活力也有一定的促进作用。     

鼎湖山马尾松、荷木混交林生态系统碳素积累和分配特征

选取鼎湖山保护区3个马尾松9Pinus massoniana),荷木（Schima superba)针阔混交林样地,研究其生态系统的碳素积累和分配特征。结果表明,鼎湖山马尾松,荷木混交林乔木尾生物量（thm^-2)为：174.41-270.11。平均227.36,且均以马尾松的生物量居多（占54.9%-84.4%)。林下层植物生物量和地表现存凋落物量（thm^-2)分别为7.41-28.28和7.06-11.56。平均14.41和9.03。三个混交林生态系统总碳贮量（thm^-2)分别为146.35,215.30和205.79。平均为189.15,其中植被层碳贮量贡献率最大,依次占62.9%,61.9%和69.9%。平均65.0%;土壤层贡献率次之,依次占34.3%,35.5%和28.5%。平均32.8%;而地表现存凋落物层的贡献最小,仅占2.8%,2.6%和1.6%。平均为2.3%。此外,本文还对该生态系统植被碳吸存潜力进行了讨论。     

蛹虫草菌丝生长研究

通过对 4种不同来源的蛹虫草菌种 ,在 5种不同配方培养基上菌丝生长情况进行了比较研究。结果表明 ,不同培养基配方对不同来源的蛹虫草菌种培养菌丝的长速和长势存在较大的差异。     

载脂蛋白E3的克隆及原核表达

载脂蛋白Ｅ(ａｐｏｌｉｐｏｐｒｏｔｅｉｎＥ ,ａｐｏＥ)由 2 99个氨基酸组成 ,分子量 34ｋＤ ,是维持人体正常脂质代谢的必需蛋白质 .它是乳糜微粒 (ＣＭ )、极低密度脂蛋白 (ＶＬＤＬ)和高密度脂蛋白 (ＨＤＬ)的组分 ,是极低密度脂蛋白受体的重要配体 ,是脂质进入细胞不可缺少的中介 .ａｐｏＥ有 3种同分异构体 :ａｐｏＥ2、ａｐｏＥ3、ａｐｏＥ4 ,分别具有不同的生理作用 .ａｐｏＥ4与血浆高胆固醇、心血管疾病和老年痴呆等疾病关联[1～ 3 ] .ａｐｏＥ2与Ⅲ型高脂血症有关 ,并对老年痴呆有防治作用[4,5] .ａｐｏＥ3是大多数健康人所具有…     

豆腐柴根提取物对小鼠非特异性免疫功能的影响

将不同剂量豆腐柴(Premna Microphylla Turcz)根提取物分别灌胃给药,通过小鼠刚果红吞噬试验,初步研究了豆腐柴根提取物对小鼠非特异性免疫功能的影响。结果表明,豆腐柴根提取物具有增强机体非特异性免疫功能的作用,其中以C组的效果最为显著。该研究为合理开发利用豆腐柴这一野生药物资源提供了科学的实验依据。