几种活血中药对ApoE缺陷小鼠动脉粥样硬化斑块的影响

目的：从形态学角度观察几种常用活血中药：丹参、赤芍、川芎、三七、桃仁和酒大黄对ApoE基因缺陷小鼠动脉粥样硬化斑块稳定性的影响。 方法： 观察主动脉根部，取4个切面，HE染色和MASSON染色，利用计算机图像分析仪测量斑块面积、血管横截面积、脂质中心面积、胶原面积，最小纤维帽厚度，计算校正斑块面积(斑块面积/血管横截面积)及脂质中心面积占斑块面积百分比，校正胶原面积(胶原面积/血管横截面积)。 结果： 与模型组比较：酒大黄组可显著降低经血管横截面积校正以后的斑块面积；丹参组、桃仁组和酒大黄组可明显减少脂质中心面积及脂质中心在AS斑块中的百分比(P＜0.05，P＜0.01)；三七组亦可明显减少脂质中心在AS斑块中的百分比(P＜0.05)。赤芍组、川芎组、桃仁组和酒大黄组的最小纤维帽较厚(P＜0.05，P＜0.01)。 结论： 提示上述活血中药可能干预ApoE缺陷小鼠动脉粥样硬化斑块的形态结构，稳定斑块，其作用与上述各味中药活血化瘀功效强弱并不完全一致。     

红藤对LPS诱导流产小鼠的保胎作用及子宫巨噬细胞的影响

目的:探讨红藤对细菌脂多糖(LPS)诱导流产小鼠的保护作用及其免疫药理机制。方法:昆明种小鼠55只,随机分为对照组(A组,10只)和实验组。实验组又分为LPS处理组(B组)、红藤处理组(C组)、红藤及LPS双处理组(D组),每组各15只。观察妊娠结果,并用酶组织化学、免疫组化、ELISA方法检测子宫非特异性酯酶阳性(α-NAE+)、CD14+、CD204+巨噬细胞和TNF-α的变化。结果:小鼠流产率在B组为100%(P<0.01),胚胎吸收率为100%(P<0.01);而D组流产率降低为13.33%,胚胎吸收率降至12.56%;孕鼠子宫α-NAE+、CD14+巨噬细胞数量,B组子宫各部位与A组相比均显著增多(P<0.01);D组环肌外侧与A组相比显著增多(P<0.01),但环肌内侧、功能层无明显变化,C、D组CD204+巨噬细胞数量与A组、B组相比均显著增多(P<0.01);子宫TNF-α含量B组与A组相比显著增加(P<0.01),而D组接近正常水平。结论:红藤可以对抗LPS所致的小鼠流产,通过影响孕鼠子宫巨噬细胞的数量、分布和亚群,抑制TNF-α的分泌,是其保护胚胎正常发育的机理之一。     

CD68和CD206阳性巨噬细胞在乳腺癌组织中的浸润及对预后影响

目的探讨乳腺癌间质中CD68~+和CD206~+肿瘤相关巨噬细胞(tumor associated macrophages,TAMs)的浸润与临床病理特征的关系及其对预后的影响。方法应用免疫组化Max Vision法检测172例乳腺癌和50例乳腺良性病变组织中CD68和CD206的表达,比较CD68~+TAMs和CD68~+/CD206~+TAMs浸润密度与乳腺癌临床病理特征的关系及其对预后的影响。结果CD68~+TAMs和CD68~+/CD206~+TAMs在乳腺癌的浸润密度均较良性病变组织升高(P均0.000 1);CD68~+/CD206~+TAMs高密度表达分别与肿瘤直径增大、淋巴结分期及临床分期增高等有关(P0.000 1,P=0.007,P0.000 1)。CD68~+/CD206~+TAMs高密度浸润组患者的无病生存率及总生存率均较低密度浸润组降低(P=0.013,P=0.003)。结论 CD68~+TAMs和CD68~+/CD206~+TAMs在乳腺癌组织中有较高的浸润密度,CD68~+/CD206~+TAMs可能与乳腺癌的发生、发展密切相关,并有望成为预测乳腺癌预后的重要潜在标志物。     

黄芪注射液对急性脑梗死CD3/HLA-DR及CD3/CD16+56的影响

细胞免疫介导的炎症反应与急性脑梗死(ACI)缺血一再灌流过程中神经元损伤、变性、程序性凋亡密切相关。黄芪具有抗菌消炎、抑制血小板凝集、增强机体耐氧、免疫调节等作用，但对ACI免疫损伤保护的研究较少。我们观察了黄芪注射液(AI)对ACI患者外周血淋巴细胞CD3／HLA-DR与CD3／CD16 56表达的影响，现报告如下。     

TNF-α对小鼠肺泡巨噬细胞CD14和清道夫受体表达的影响

本研究旨在通过观察TNF-α对肺泡巨噬细胞(AM)清道夫受体(SR)、CD14表达的影响，探讨TNF-α在内毒素血症时AM由早期防御性(清除、灭活内毒素)向后期效应性(释放大量的致炎与抗炎因子)转变中的作用。分离培养小鼠AM，以不同剂量TNF-α(0，0．001，0．01，0．1，1，10，100μg／L)刺激细胞16h或以100μg／L TNF-α刺激细胞不同时间(0，2，4，6，8，12，16，24h)，采用免疫细胞化学及RT-PCR方法观察SR、CD14表达变化。结果显示，以低至0．1μg／L TNF-α刺激16h或100μg／L TNF-α刺激6h以上就能显著增强CD14并抑制SR蛋白的表达，实验同时显示，CD14mRNA、SRmRNA表达也显著增强，SRmRNA表达变化与SR蛋白表达趋势不一致。研究结果提示，TNF-α能通过增强CD14蛋白表达并抑制SR蛋白表达而在内毒素血症时小鼠AM由免疫防御型向炎症效应型转变中发挥重要作用。     

6种活血中药及芎芍胶囊对ApoE基因缺陷小鼠动脉粥样硬化斑块胶原沉积及其代谢的影响

目的：探讨6种常用活血中药及芎芍胶囊对于ApoE基因缺陷小鼠动脉粥样斑块胶原沉积及其代谢的影响。方法：6-8周龄ApoE基因缺陷鼠100只，雌雄各半，体重18-20g。基础饲料喂养6-8周后，改为“西方膳食类”饲料（含脂肪21％（wt／wt）、胆固醇0.15％（wt／wt））继续喂养13周，随机分成9组：模型组、对照组（辛伐他汀）、丹参组、赤芍组、川芎组、三七组、酒大黄组、桃仁组、和芎芍胶囊组，灌胃给药13周。无菌条件下取出心脏和主动脉，10％甲醛固定，脱水，常规石蜡包埋切片。苦味酸-天狼星红染色，在偏振光下，观察Ⅰ、Ⅲ型胶原；免疫组化染色检测主动脉根部动脉粥样斑块及周围血管壁MMP-1、TIMP-1，利用IPP图像分析软件计算Ⅰ、Ⅲ型胶原含量变化。IPP图像分析软件计算表达阳性面积及阳性面积占总面积比。     

胰岛CD68、CD22阳性细胞表达及血清IL-12、IL-4水平的改变对1型糖尿病小鼠的影响

目的探讨1型糖尿病(T1DM)小鼠胰岛表达CD68、CD22的改变,血清白介素-12(IL-12)、白介素-4(IL-4)水平的变化及可能的机制。方法正常雄性C57BL/6J小鼠104只,随机分为正常对照组、盐水对照组及实验组。小剂量多次注射链脲佐菌素(MLD-STZ)建立1型糖尿病小鼠模型,分别于第3、7、10、14、21、28天测空腹血糖,取胰尾组织及血清,通过免疫组化SABC法、酶联免疫吸附法(ELISA)及形态计量法进行研究。结果与正常及盐水对照组相比较,实验组小鼠胰岛数目减少,胰岛面积减小。实验组小鼠胰岛CD68阳性细胞的面数密度(NA)于第3天起明显高于正常对照组和盐水对照组(P0.01),但第10天的NA在实验组相对较低。实验组胰岛CD22阳性细胞的NA值于第3天即高于正常对照组和盐水对照组(P0.01),至第7天最高。血清ELISA结果显示,实验组小鼠血清IL-12水平自第7天开始增高(P0.05);而血清IL-4水平与正常及盐水对照组相比有所降低,差异有统计学意义(P0.05)。结论 CD68及CD22阳性细胞在T1DM早期即浸润胰岛,有可能通过抗原递呈等作用促进了T1DM的发生。实验组小鼠血清IL-12增高,IL-4减少,提示Th1/Th2细胞失衡,推测是引发T1DM的重要因素之一。     

CD14抑制肽对内毒素诱导U937细胞表达TNF-α的影响

目的 观察CD14抑制肽(CD14 inhibitory peptide,CD14-IP)对内毒素诱导的U937细胞表达TNF-α的影响.方法 U937细胞用佛波脂(PMA)诱导成熟后分5组:正常对照组、LPS组、高剂量抑制肽组、中剂量抑制肽组和低剂量抑制肽组.LPS组给予终浓度为100 ng/mL的LPS和100 ng/mL的LBP,高、中和低剂量抑制肽组除给予LPS和LBP外,分别给予终浓度为10 μg/mL、1.0 μg/mL和0.1 μg/mL的CD14-IP.ELISA测定细胞培养上清NNF-α的浓度.进一步观察不同时间应用CD14-IP(1.0 μg/mL)对LPS诱导U937细胞TNF-α和NTF-α mRNA表达的影响.用RT-PCR测定细胞TNF-α mRNA的表达.结果 LPS组和抑制肽各组TNF-α浓度较正常组明显增高(P<0.05),高、中剂量抑制肽组TNF-α水平比LPS组明显降低(P<0.05),高、中剂量抑制肽组之间TNF-α浓度无明显差别(P>0.05),低剂量抑制肽组TNF-α浓度与LPS组无统计学差异(P>0.05).不同时间应用CD14-IP时,CD14-IP早期应用对TNF-α和TNF-α mRNA的表达抑制作用显著.结论 CD14-IP能显著减少LPS诱导的U937细胞TNF-α和TNF-α mRNA的表达,早期应用效果较好,可能对LPS所致急性肺损伤有保护作用.     

胃癌患者PBMCs中CD4+CD25+T细胞体外增殖及对CD4+CD25-T细胞增殖的影响

目的:探讨胃癌患者外周血单个核细胞（PBMCs）中的CD4+CD25+T细胞体外增殖及对CD4+CD25-T细胞增殖的影响。 方法:以免疫磁性分离方法 （MACS）分选出胃癌患者外周血单个核细胞中的CD4+CD25+T及CD4+CD25-T细胞后，用流式细胞仪分析细胞的纯度及活力；再以小鼠抗人CD3单抗、小鼠抗人CD28单抗及rh IL-2作为共刺激因子，观察与CD4+CD25-T细胞共培养时，CD4+CD25+T细胞对CD4+CD25-T细胞增殖的抑制效应。 结果:（1）分选后健康对照组及胃癌患者PBMC 中CD4+CD25+ T细胞纯度分别为83.8%±1.84%、84.13%±2.77％，两者相比，无显著差异（P＞0.05）;(2)经MACS 分选后正常对照组与胃癌患者CD4+CD25+ T细胞活力分别为98.52%±0.72%、97.80%±0.95%，两者相比，无显著差异（P＞0.05）；(3)无论是健康对照还是胃癌患者的CD4+CD25+T均具有明显抑制效应性T细胞如CD4+CD25-T细胞的增殖，随着CD4+CD25+T细胞数的增加，这种抑制增殖的能力也相应增加，当CD4+CD25+∶〖KG-*2〗CD4+CD25-T达 1∶〖KG-*2〗1时，抑制率最大达到50％。 结论:MACS分选法能够分选出高纯度及活力的CD4+CD25+T细胞，分选后CD4+CD25+T细胞在体外均能抑制CD4+CD25-T细胞增殖，且这种抑制效应呈一定效靶比关系。     

Comparison of astrocytic and myocytic metabolic dysregulation in apolipoprotein E deficient and human apolipoprotein E transgenic mice

The accumulation of tubular aggregates in type II skeletal muscle fibres and fibrillo-granular inclusions in hippocampal protoplasmic astrocytes are characteristic lesions of apolipoprotein E deficient mice. Moreover these inclusions reacted immunocytochemically with an antibody specific to fragment 17-24 of the published sequence of Alzheimer's amyloid peptide. In an effort to evaluate the role of apolipoprotein E in the formation of these abnormal structures, we examined the tibialis anterior muscle and the hippocampus of several groups of animals including: (i) apolipoprotein E "knockout" mice which had been whole body irradiated with 1200 rads and bone marrow replenished with apolipoprotein E sufficient marrow; and (ii) three transgenic murine strains that had been genetically engineered to express either human apolipoprotein E2, E3 or E4 protein on an apoE deficient background. The results of this study showed that the presence of murine apolipoprotein E (even in subnormal levels in the serum) in irradiated bone marrow replenished mice and in all three (E2, E3 or E4) human apoE transgenic strains was sufficient to prevent the aggregation of sarcoplasmic tubules in the tibialis anterior type II muscle fibres. Similarly apolipoprotein E "knockout" bone marrow replenished mice and all three transgenic strains expressing the different human apolipoprotein E alleles reduced the number of the astrocytic inclusions in the hippocampus to levels not significantly different to those observed in control C57Bl6J animals.The data obtained in this study indicate that neurological and neuromuscular abnormalities found in apoE deficient mice are reversed when apoE protein is replaced in the circulation, either by bone marrow transplantation of normal apoE sufficient marrow, or by gene therapy with the apoE gene, albeit of human origin and irrespective of the allele used.     

Cardiovascular functional phenotypes and pharmacological responses in apolipoprotein E deficient mice

This is an overview of recent findings, mainly from our laboratory, describing the cardiovascular functional phenotypes and pharmacological responses in mice genetically deficient in apolipoprotein E (apoE-KO). ApoE-KO mice are hyperlipidemic and spontaneously develop atherosclerosis. We have detected several new cardiovascular functional phenotypes in apoE-KO mice: hyperglycemia, age-dependent aortic stiffening, cardiac hypertrophy and increased cardiac output. Angiotensin II (Ang II) promoted vascular inflammation and atherosclerosis, increased vascular stiffness, and induced abdominal aortic aneurysm (AAA) in apoE-KO mice, in which activation of NF-kappaB mediated pro-inflammatory genes plays an important role. Inhibition of nitric oxide (NO) synthesis with N(omega)-nitro-L-arginine methyl ester (L-NAME) significantly inhibited NO-mediated vascular responses and accelerated atherosclerosis in apoE-KO mice, supporting a protective role of NO against atherosclerosis. Estrogen attenuated atherosclerosis in apoE-KO mice, even in those with atherosclerosis being accelerated by Ang II, hyperglycemia, or L-NAME, demonstrating an anti-atherosclerotic effect of estrogen. Simvastatin paradoxically increased lipid and atherosclerosis in apoE-KO mice, but it decreased lipid and atherosclerosis in LDLR-KO mice, indicating that anti-atherosclerotic effect of simvastatin requires the presence of an intact apoE.     

3,4,5,6-四羟基(口山)酮抑制ApoE基因缺陷小鼠动脉粥样硬化形成及机制

目的：研究3,4,5,6-四羟基口山酮抑制ApoE基因缺陷小鼠动脉粥样硬化(AS)形成的机制及其与内源性一氧化氮合酶(NOS)抑制剂非对称性二甲基精氨酸(ADMA)的关系。方法：32只小鼠分为4组(n=8):正常对照组(C57BL/6J小鼠，等体积溶媒灌胃)；模型组(ApoE基因缺陷小鼠，等体积溶媒灌胃)；低剂量口山酮组［ApoE基因缺陷小鼠，口山酮10mg/（kg·d）灌胃］；高剂量口山酮组［ApoE基因缺陷小鼠，口山酮20mg/（kg·d）灌胃］；检测主动脉脂质斑块面积、血脂、红细胞变形性和血浆ADMA的水平。结果：与模型组比较，口山酮可以显著降低血浆ADMA水平和斑块面积，降低血浆总胆固醇(TC)、血浆总甘油三酯(TG)、血浆低密度胆固醇(LDL-C)，显著升高血浆高密度胆固醇(HDL-C)和改善红细胞变形性。结论：3,4,5,6-四羟基口山酮具有抗AS作用, 其作用机制与改善脂质代谢和降低血浆ADMA的水平有关。     

Regional difference of lipid distribution in brain of apolipoprotein E deficient mice

According to recent knowledge, apolipoprotein E (apo E) plays a significant role in the homeostasis of intracellular cholesterol level in various tissues. Apo E deficient mice develop hyperlipidemia, and suffer from atherosclerosis in extracerebral blood vessels and neurodegeneration in the central nervous system. Furthermore, Walker et al. (Am. J. Path., 1997;151:1371–1377) demonstrated cerebral xanthomas of various sizes in the brain of apo E deficient mice. In the present study, it is illustrated that in the homozygous apo E deficient mice of advancing age, a great number of foamy macrophages extravasate from microvessels in thalamus and fimbria hippocampi, and scatter in the perivascular regions and migrate toward the ependyma, fimbria hippocampi, hippocampus, and thalamus. Here, it must be pointed out that under hyperlipidemia, although foamy macrophages made clusters in the perivascular region, the cerebral microvessels did not develop atherosclerosis. On the other hand, in the other cerebral regions such as cerebral cortex, caudoputamen, globus pallidus, and substantia nigra, macrophages did not appear and microvessels retained normal shapes, but the fluorescent granular perithelial (in short, FGP) cells accompanied by these vessels contained a certain amount of lipids. That is, in the cerebral cortex and caudoputamen, lipid components are detected in FGP cells and microglia, while in the globus pallidus and substantia nigra, they are mainly localized in astrocytes. The reason why the astrocytes in such defined regions contain, specifically, a high quantity of lipid components remains unsettled. Axonal degenerations are often represented in thalamus, globus pallidus, and substantia nigra. On the other hand, in the specimens of Wild‐type mice, lipid components were observed only in FGP cells, and the vascular architecture took a normal profile. Any lipid laden macrophages and the axonal degenerations could not be detected through the cerebral parenchyma. Furthermore, it is also a noticeable finding that immunohistochemically, the FGP cells express a positive reaction against the antibody of apo E in the Wild‐type mice, but those of homozygous apo E deficient mice are immunonegative. FGP cells are not only provided with the scavenger receptor, but also contribute to the lipid metabolism in the brain. Anat Rec 256:165–176, 1999. © 1999 Wiley‐Liss, Inc.     

Regional difference of lipid distribution in brain of apolipoprotein E deficient mice.

According to recent knowledge, apolipoprotein E (apo E) plays a significant role in the homeostasis of intracellular cholesterol level in various tissues. Apo E deficient mice develop hyperlipidemia, and suffer from atherosclerosis in extracerebral blood vessels and neurodegeneration in the central nervous system. Furthermore, Walker et al. (Am. J. Path., 1997;151:1371-1377) demonstrated cerebral xanthomas of various sizes in the brain of apo E deficient mice. In the present study, it is illustrated that in the homozygous apo E deficient mice of advancing age, a great number of foamy macrophages extravasate from microvessels in thalamus and fimbria hippocampi, and scatter in the perivascular regions and migrate toward the ependyma, fimbria hippocampi, hippocampus, and thalamus. Here, it must be pointed out that under hyperlipidemia, although foamy macrophages made clusters in the perivascular region, the cerebral microvessels did not develop atherosclerosis. On the other hand, in the other cerebral regions such as cerebral cortex, caudoputamen, globus pallidus, and substantia nigra, macrophages did not appear and microvessels retained normal shapes, but the fluorescent granular perithelial (in short, FGP) cells accompanied by these vessels contained a certain amount of lipids. That is, in the cerebral cortex and caudoputamen, lipid components are detected in FGP cells and microglia, while in the globus pallidus and substantia nigra, they are mainly localized in astrocytes. The reason why the astrocytes in such defined regions contain, specifically, a high quantity of lipid components remains unsettled. Axonal degenerations are often represented in thalamus, globus pallidus, and substantia nigra. On the other hand, in the specimens of Wild-type mice, lipid components were observed only in FGP cells, and the vascular architecture took a normal profile. Any lipid laden macrophages and the axonal degenerations could not be detected through the cerebral parenchyma. Furthermore, it is also a noticeable finding that immunohistochemically, the FGP cells express a positive reaction against the antibody of apo E in the Wild-type mice, but those of homozygous apo E deficient mice are immunonegative. FGP cells are not only provided with the scavenger receptor, but also contribute to the lipid metabolism in the brain.     

In vivo imaging of atherosclerotic plaques in apolipoprotein E deficient mice using nonlinear microscopy

Structural proteins such as elastin and collagen can be readily imaged by using two-photon excitation and second-harmonic generation microscopic techniques, respectively, without physical or biochemical processing of the tissues. This time- and effort-saving advantage makes these imaging techniques convenient for determining the structural characteristics of blood vessels in vivo. Fibrillar collagen is a well-known element involved in the formation of atherosclerotic lesions. It is also an important component of the fibrous cap responsible for structural stability of atherosclerotic plaques. High resolution in vivo microscopic imaging and characterization of atherosclerotic lesions in animal models can be particularly useful for drug discovery. However, it is hindered by the limitations of regular microscope objectives to gain access of the tissues of interest and motional artifacts. We report a technique that facilitates in vivo microscopic imaging of carotid arteries of rodents using conventional microscope objectives, and at the same time avoids motional artifacts. As a result, collagen, elastin, leukocytes, cell nuclei, and neutral lipids can be visualized in three dimensions in live animals. We present and discuss in vivo imaging results using a flow cessation mouse model of accelerated atherosclerosis.     

Anti-oxidant and anti-inflammatory effects of apigenin in a rat model of sepsis: an immunological,biochemical, and histopathological study

Objective: We hypothesize that apigenin may inhibit some cellular process of sepsis-induced spleen injury and simultaneously improve inflammation and oxidative stress. Therefore, the aim of this study was to investigate the potential protective effects of apigenin in a polymicrobial sepsis rat model of by cecal ligation and puncture.

Materials and methods: 64 female Wistar albino rats were divided into 8 groups. The pro-inflammatory (tumor necrosis factor-alpha, interleukin-6, and interleukin-1-beta) and anti-inflammatory (tumor growth factor-beta and interleukin-10) cytokine levels were measured by enzyme-linked immunosorbent assay. CD3, CD68, and nuclear factor kappa B (NF-κB) positivity rates were detected by immunohistochemical methods. Oxidative stress parameters were measured by tissue biochemistry.

Results: Sepsis caused a significant increase in TNF-alpha, IL-1-beta, IL-6, and TGF-beta levels whereas it reduced IL-10 level. Additionally, it led to an increase in CD3, CD68, and NF-κB positivity rates as well as oxidative stress parameters levels. However, apigenin inhibited the inflammation process, increased the IL-10 level and normalized the oxidative stress parameters.

Discussion and conclusion: Pretreatment with apigenin results in a significant reduction in the amount of inflammatory cells. The beneficial effect of apigenin on spleen injury also involved inhibition of NF-κB pathway, suppression of proinflammatory cytokines, and induction of anti-inflammatory cytokine production. Additionally, it led to a decrease in oxidative stress in spleen tissue. Taking everything into account, apigenin may be an alternative therapeutic option for prevention of sepsis-induced organ.     

ILC2 transfers to apolipoprotein E deficient mice reduce the lipid content of atherosclerotic lesions

The use of immunodeficient mice transplanted with human hematopoietic stem cells is an accepted approach to study human-specific infectious diseases such as HIV-1 and to investigate multiple aspects of human immune system development. However, mouse and human are different in sialylation patterns of proteins due to evolutionary mutations of the CMP-N-acetylneuraminic acid hydroxylase (CMAH) gene that prevent formation of N-glycolylneuraminic acid from N-acetylneuraminic acid. How changes in the mouse glycoproteins’ chemistry affect phenotype and function of transplanted human hematopoietic stem cells and mature human immune cells in the course of HIV-1 infection are not known. We mutated mouse CMAH in the NOD/scid-IL2Rγc−/− (NSG) mouse strain, which is widely used for the transplantation of human cells, using the CRISPR/Cas9 system. The new strain provides a better environment for human immune cells. Transplantation of human hematopoietic stem cells leads to broad B cells repertoire, higher sensitivity to HIV-1 infection, and enhanced proliferation of transplanted peripheral blood lymphocytes. The mice showed no effect on the clearance of human immunoglobulins and enhanced transduction efficiency of recombinant adeno-associated viral vector rAAV2/DJ8. NSG-cmah−/− mice expand the mouse models suitable for human cells transplantation, and this new model has advantages in generating a human B cell repertoire. This strain is suitable to study different aspects of the human immune system development, provide advantages in patient-derived tissue and cell transplantation, and could allow studies of viral vectors and infectious agents that are sensitive to human-like sialylation of mouse glycoproteins.     

Differential neurotrophic effects of apolipoprotein E in aged transgenic mice.

The present study seeked to determine whether the neurodegenerative and cognitive alterations in aged apolipoprotein E-deficient mice are differentially reversed by transgenic overexpression of human apolipoprotein-E3 vs. apolipoprotein-E4 in the background of deficient endogenous apolipoprotein E. These studies showed dendritic alterations in pyramidal neurons of apolipoprotein-E4 transgenic mice, similar to the ones observed in apolipoprotein E-deficient mice. However, these mice had a preserved density of synaptophysin-immunoreactive presynaptic terminals. In contrast, mice overexpressing apolipoprotein-E3 showed no synapto-dendritic alterations. Analysis of behavioral performance in the Morris water maze showed that while apolipoprotein E-deficient mice performed poorly, overexpression of apolipoprotein-E3 and, to a lower extent apolipoprotein-E4, resulted in an improved performance. This study supports the contention that, compared with apolipoprotein-E4, apolipoprotein-E3 might have a greater neurotrophic in vivo effect in aged mice.     

Predisposition to atherosclerosis and aortic aneurysms in mice deficient in kinin B1 receptor and apolipoprotein E

Kinin B1 receptor is involved in chronic inflammation and expressed in human atherosclerotic lesions. However, its significance for lesion development is unknown. Therefore, we investigated the effect of kinin B1 receptor deletion on the development of atherosclerosis and aortic aneurysms in apolipoprotein E-deficient (ApoE^−/−) mice. Mice deficient both in ApoE and in kinin B1 receptor (ApoE^−/−-B₁^−/−) were generated and analyzed for their susceptibility to atherosclerosis and aneurysm development under cholesterol rich-diet (western diet) and angiotensin II infusion. Kinin B1 receptor messenger RNA (mRNA) expression was significantly increased in ApoE^−/− mice after Western-type diet. Although no difference in serum cholesterol was found between ApoE^−/−-B₁^−/− and ApoE^−/− mice under Western-type diet, aortic lesion incidence was significantly higher in ApoE^−/−-B₁^−/− after this treatment. In accordance, we observed increased endothelial dysfunction in these mice. The mRNA expression of cyclic guanosine monophosphate-dependent protein kinase I, CD-11, F4/80, macrophage colony-stimulating factor, and tumor necrosis factor-alpha were increased in the aorta of double-deficient mice following Western-type diet, whereas the levels of peroxisome proliferator-activated receptor gamma protein and the activity of matrix metalloproteinase-9 activity were decreased. In addition to the increased atherosclerotic lesions, the lack of kinin B₁ receptor also increased the incidence of abdominal aortic aneurysms after angiotensin II infusion. In conclusion, our results show that kinin B₁ receptor deficiency aggravates atherosclerosis and aortic aneurysms under cholesterolemic conditions, supporting an antiatherogenic role for the kinin B₁ receptor.