CXCL12 Retargeting of an Oncolytic Adenovirus Vector to the Chemokine CXCR4 and CXCR7 Receptors in Breast Cancer

Breast cancer is the most frequently diagnosed cancer in women under 60, and the second most diagnosed cancer in women over 60. While significant progress has been made in developing targeted therapies for breast cancer, advanced breast cancer continues to have high mortality, with poor 5-year survival rates. Thus, current therapies are insufficient in treating advanced stages of breast cancer;new treatments are sorely needed to address the complexity of advanced-stage breast cancer. Oncolytic virotherapy has been explored as a therapeutic approach capable of systemic administration, targeting cancer cells, and sparing normal tissue. In particular, oncolytic adenoviruses have been exploited as viral vectors due to their ease of manipulation, production, and demonstrated clinical safety profile. In this study, we engineered an oncolytic adenovirus to target the chemokine receptors CXCR4 and CXCR7. The overexpression of CXCR4 and CXCR7 is implicated in the initiation, survival, progress, and metastasis of breast cancer. Both receptors bind to the ligand, CXCL12 (SDF-1), which has been identified to play a crucial role in the metastasis of breast cancer cells. This study incorporated a T4 fibritin protein fused to CXCL12 into the tail domain of an adenovirus fiber to retarget the vector to the CXCR4 and CXCR7 chemokine receptors. We showed that the modified virus targets and infects CXCR4- and CXCR7-overexpressing breast cancer cells more efficiently than a wild-type control vector. In addition, the substitution of the wild-type fiber and knob with the modified chimeric fiber did not interfere with oncolytic capability. Overall, the results of this study demonstrate the feasibility of retargeting adenovirus vectors to chemokine receptor-positive tumors.

Relationship between expression of chemokine receptors CCR3, CCR5 and CXCR3 on CD4^＋ T cells and spontaneous abortion in mice

Background Previous studies have shown that local immune cells in the feto-maternal interface are recruited from peripheral blood, and that chemokines and their receptors play an initial and key role in this recruitment process. In this study, we aimed to determine whether spontaneous abortion is associated with the expression of chemokine receptors CCR3, CCR5, and CXCR3 on CD4^＋ T cells. Methods Peripheral blood, spleen, and thymus were collected from the spontaneous abortion mouse model CBA/JxDBA/2 （SA group, n=14）, the normal pregnant mouse model CBA/JxBALB/c （NP group, n=13）, and normal non-pregnant CBA/J mice （NNP group, n=11）. The number of chemokine receptors CCR3, CCR5, and CXCR3 expressed on CD4^＋ T cells was measured by double-label flow cytometry （FCM） method. Results In peripheral blood, the SA group had significantly lower CCR3 expression （P 〈0.01） and higher CCR5 and CXCR3 expression （P 〈0.01） on CD4^＋ T cells than did the NP group. But comparing these chemokines between the SA and NNP groups, there was no significant difference （P 〉0.05）. In spleen, the SA group expressed significantly lower CCR3 expression （P 〈0.01） and higher CCR5 and CXCR3 expression （P 〈0.05） on CD4^＋ T cells than did the NP group. When compared with the NNP group, the SA group had significantly higher CCR3 expression （P 〈0.01）, but was not statistically different with regards to the other two chemokines （P 〉0.05）. In thymus, the SA group had significantly lower CCR3 expression （P 〈0.05） and higher CXCR3 expression （P 〈0.05） on CD4^＋ T cells than the NP group, with no significant difference in CCR5 expression （P 〉0.05）. Compared with the NNP group, the SA group had higher CCR3 expression （P 〈0.01）, but there was no statistical difference in CXCR3 and CCR5 expression （P 〉0.05） between the two groups. Conclusion The abnormal expression of CCR3, CCR5 and CXCR3 on CD4^＋ T cells may play an important role in the pathogenesis of spontaneous abortion.

Association between polymorphism of MC4R rs489693 gene and disorder of glucose and lipid metabolism in schizophrenia patients treated with olanzapine

Objective:To investigate the effect of Melanocortin four receptor,MC4R rs489693 polymorphism on glucose and Lipid metabolism in schizophrenia patients treated with Olanzapine for 12 weeks.Methods:171 patients with schizophrenia were divided into AA Group(N=12),AC group(N=59)and CC Group(N=100)according to the polymorphism of MC4R gene at 489693 locus detected by DNA sequencing.Blood Glucose and lipid levels were measured before and 12 weeks after treatment,the differences of variables among the 3 groups were compared,and the incidence of glucose and lipid abnormalities after treatment was statistically analyzed.Results:After 12 weeks of treatment,the net increase of blood glucose in AA group was greater than that in CC group(P<0.05),and the net increase of cholesterol and triglyceride in AA group was greater than that in AC group and CC group(all P<0.05),and the incidence of Blood Glucose and at least one dyslipidemia in AA Group was higher than that in AC and CC group(all P<0.01).Conclusion:The rs489693 gene polymorphism of MC4R gene is related to the disorder of glucose and lipid metabolism in schizophrenia treated with olanzapine.

缺氧诱导因子1α、血管内皮生长因子及CXCR4受体在甲状腺乳头状癌中的表达及与肿瘤转移的关系

甲状腺乳头状癌（papillary thyroid carcinoma,PTC）占甲状腺恶性肿瘤的80%以上,大多数生长缓慢预后良好,但仍有一部分表现为明显的侵袭性行为,包括淋巴结转移、远处转移、治疗抵抗及致死性,寻找甄别及治疗该类PTC的方法是临床亟待解决的问题之一[1]。缺氧诱导因子1α（hypoxia-inducible factor 1,alpha Subunit,HIF-1α）、血管内皮生长因子（vascular endothelial growth factor,VEGF）及CXCR4受体在体内外试验中均证实参与肿瘤的侵袭与转移过程，

Correlation of the changes of TLR4/NF-κB pathway function in intrauterine adhesion tissue with the characteristics of cytokine secretion and collagen metabolism

Objective:To study the correlation of the changes of Toll-like receptor 4 (TLR4) / nuclear factorκB (NF-κB) pathway function in intrauterine adhesion (IUAs) tissue with the characteristics of cytokine secretion and collagen metabolism.Methods:The patients with IUAs who were treated in our hospital between February 2015 and March 2018 were selected as the IUAs group, and the patients who underwent hysteroscopy due to infertility and were pathologically confirmed to have normal endometrium during the same period were selected as the control group. The expression levels of TLR4/NF-κB pathway molecules and collagen metabolism genes as well as the contents of cytokines and collagen metabolism markers in the adhesion tissues of IUAs group and the normal endometrial tissue of control group were measured.Results: TLR4, NF-κB, a disintegrin and metalloproteinase 15 (ADAM15), ADAM17, matrix metalloproteinase 9 (MMP9) and plasminogen activator inhibitor 1 (PAI-1) mRNA expression as well as transforming growth factor-β1 (TGF-β1), Smad2/3, insulin-like growth factor 1 (IGF-1), IGF-1 receptor (IGF-1R), basic fibroblast growth factor (bFGF), periostin/osteoblast-specific factor 2 (Postn), type I collagen (Col-I) and actin-α (α-SMA) contents in the adhesion tissues of IUAs group were significantly higher than those of control group while urokinase-type plasminogen activator (uPA) mRNA expression was significantly lower than that of control group;TLR4 and NF-κB mRNA expression were positively correlated with TGF-β1, Smad2/3, IGF-1, IGF-1R, bFGF, Postn, Col-I,α-SMA, ADAM15, ADAM17, MMP9 and PAI-1, and negatively correlated with uPA.Conclusion:The excessive activation of TLR4/NF-κB pathway in IUAs is associated with the cytokine secretion and collagen metabolism abnormalities.

补髓生血颗粒剂对慢性再生障碍性贫血患者骨髓造血祖细胞c-kit、CXCR_4受体表达的影响

目的 :探讨补髓生血颗粒剂对慢性再生障碍性贫血 (CAA)患者骨髓造血祖细胞c -kit、CXCR4受体表达水平的影响。方法 :采用免疫荧光法及流式细胞技术分别对使用该颗粒剂治疗前后的CAA患者骨髓造血祖细胞c-kit、CXCR4 受体表达水平进行检测 ,并与再障生血片对照组及正常组作比较研究。结果 :补髓生血颗粒剂组临床疗效与再障生血片组疗效存在显著差异(P <0 .0 5 ) ;两组疗前c -kit受体表达高于正常组 ,治疗后均表达下调 ;补髓生血颗粒剂组治疗后CXCR4 受体表达与再障生血片组存显著性差异(P <0 .0 5 )。结论 :慢性再障骨髓造血衰竭并非c -kit受体低表达所致 ;补髓生血颗粒剂在临床疗效与CXCR4 受体表达上调作用上优于再障生血片组 。

母体血清CXCL12、CXCR4、VEGF及PLGF的水平与子痫前期及其合并FGR的相关性研究

目的通过检测孕产妇血清中CXC类趋化因子配体12(CXCL12)、CXC类趋化因子受体4(CXCR4)、血管内皮生长因子(VEGF)、胎盘生长因子(PLGF)的水平,探索CXCL12、CXCR4联合VEGF、PLGF在胎盘形成过程中所起的作用,进一步探讨与子痫前期(PE)及其合并胎儿生长受限(FGR)发病的关系。方法选择2018年5月至2019年5月在安徽省妇幼保健院住院分娩的孕妇为研究对象。诊断子痫前期患者103例为子痫前期组(PE组),其中子痫前期不合并胎儿生长受限患者66例(PEFGR组),子病前期合并胎儿生长受限患者37例(PE+FGR组),选择同时期住院分娩的正常产妇51例为正常对照组(N组)。采用酶联免疫吸附法(ELISA)检测母体血清中CXCL12、CXCR4、VEGF及PLGF的水平,并进行统计学分析。结果PE组母体血清中CXCL12、CXCR4、VEGF、PLGF的水平均较N组下降,差异均具有统计学意义(t=12.9、22.2、15.9、10.3,均P<0.05)。N组、PE-FGR组、PE+FGR组母体血清中CXCL12、CXCR4、VEGF、PLGF的水平差异均具有统计学意义(F=113.0、339.9、149.5、64.1,均P<0.05),进一步两两比较后发现,N组、PE-FGR组、PE+FGR组的四个血清指标均依次下降,差异具有统计学意义(均P<0.05)。结论CXCL12、CXCR4联合VEGF、PLGF在胎盘形成过程中起到重要作用;母体血清中CXCL12、CXCR4、VEGF、PLGF水平的降低可能与子痫前期及其合并FGR的发病有着重要关系。

The prognostic value of C-X-C motif chemokine receptor 4 in patients with sporadic malignant peripheral nerve sheath tumors

Background: Recent studies indicate that C-X-C motif chemokine receptor 4(CXCR4) and its ligand, C-X-C motif chemokine ligand 12(CXCL12), stimulate expression of the cell cycle regulatory protein Cyclin D1 in neurofibromatosis 1-associated malignant peripheral nerve sheath tumor(MPNST) cells and promote their proliferation. In this study, we measured the expression of CXCR4, CXCL12, and Cyclin D1 proteins in sporadic MPNST tissues from Chinese patients and investigated their prognostic values.Methods: CXCR4, CXCL12, and Cyclin D1 protein expression in samples from 58 Chinese patients with sporadic MPNST was assessed with immunohistochemical staining.Their prognostic values were evaluated with Kaplan-Meier analysis and a log-rank test. Multivariate Cox regression analysis was used to identify independent prognostic factors.Results: High expression of CXCR4, CXCL12, and Cyclin D1 was observed in 19(32.8%), 32(55.2%), and 16(27.6%)samples, respectively. CXCR4 expression was positively correlated with CXCL12 expression(r = 0.334, P = 0.010) and Cyclin D1 expression(r = 0.309, P = 0.018). Patients with high CXCR4 expression showed longer overall survival than those with low CXCR4 expression(χ~2 = 4.642, P = 0.031).Conclusion: High CXCR4 expression may define a specific subtype of sporadic MPNST with favorable prognosis.

Expressions of CCR7 and CXCR4 Are Associated with Differentiation in Gastrointestinal Cancer

Purpose: The chemokine receptors CCR7 and CXCR4 have been shown to play an important role in cancer invasion and metastasis. This study was aimed to investigate CCR7 and CXCR4 expressions and evaluate the association between their expressions and the clinicopathological features in gastrointestinal cancer. Method: 27 paired tissue samples from patients who had curative surgery for gastrointestinal cancer were obtained. Quantitative real-time PCR, immunochemistry assay and western blot analysis were carried out to investigate the expressions of CCR7, CXCR4 expressions in gastrointestinal cancer. Results: The cancer tissues expressed significant higher level of CCR7 (P = 0.000) and CXCR4 (P = 0.000) protein than the adjacent normal mucosa. Expressions of CCR7 (P = 0.002) and CXCR4 (P = 0.003) protein in cancer tissues exhibited significant correlation with differentiation in gastrointestinal cancer. Conclusion: Expressions of CCR7 and CXCR4 protein were associated with differentiation in gastrointestinal cancer. CCR7 and CXCR4 may be predictive factors for poor prognosis in patients with gastrointestinal cancer.

银杏叶提取物基于TLR4/NF-κB通路对骨质疏松大鼠的干预效果及作用机制

目的分析银杏叶提取物基于TLR4/NF-κB通路对骨质疏松大鼠的干预效果及作用机制。方法选取周龄在4~6周的SPF健康雌性SD大鼠40只,随机分对照组(A组)、骨质疏松组(B组)、阳性药物己烯雌酚组(C组)和银杏叶提取物(D组),适应性饲养1周后,第2周进行建模,饲养3个月后起给予药物干预,C组大鼠每日给予己烯雌酚0.05 mg·kg^(-1)·d^(-1),D组大鼠给予银杏叶提取物,以200 mg·kg^(-1)·d^(-1)的剂量给药,给药方式均采用灌胃给药。C组、D组进行药物干预6周,与此同时,A组、B组给予10 mL·kg^(-1)·d^(-1)0.9%氯化钠溶液灌胃。给药周期满6周后,麻醉大鼠成功,处死大鼠,取出股骨组织,检测并观察各组大鼠碱性磷酸酶(alkaline phosphatase,ALP)、骨保护素(osteoprotegerin,OPG)、25-羟基维生素D 3(25-hydroxyvitamin D_(3),25OHD)、抗酒石酸盐酸性磷酸酶(Tartrate-resistant acid phosphatase,TRACP)、钙离子(calcium ion,Ca^(2+))、镁离子(magnesian ion,Mg^(2+))、超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)、过氧化氢酶(catalase,CAT)水平及toll样受体4(toll-like receptoR4,TLR4)、核因子κB(nucleaRfactoRkappa-B,NF-κB)蛋白表达。结果与A组比较,B组、C组、D组ALP、TRACP、MDA、TLR4、NF-κB水平升高,OPG、25OHD、Ca^(2+)、Mg^(2+)、SOD、GSH-Px、CAT水平降低,差异有统计学意义(P<0.05);与B组比较,C组、D组ALP、TRACP、MDA、TLR4、NF-κB水平降低,OPG、25OHD、Ca^(2+)、Mg^(2+)、SOD、GSH-Px、CAT水平升高,差异有统计学意义(P<0.05);与C组比较,D组ALP、TRACP、MDA、TLR4、NF-κB水平降低,OPG、25OHD、Ca^(2+)、Mg^(2+)、SOD、GSH-Px、CAT水平升高,差异有统计学意义(P<0.05)。结论银杏叶提取可能会通过抑制TLR4/NF-κB通路活化来改善骨质疏松大鼠骨代谢水平及氧化应激反应,影响破骨细胞吸收,同时会调节机体Ca^(2+)、Mg^(2+)水平,未来对临床骨质疏松的治疗有重要意义。

多囊卵巢综合征患者血清TLR4表达水平与糖脂代谢的关系

目的探究多囊卵巢综合征(PCOS)患者血清Toll样受体4(TLR4)表达水平与糖脂代谢的相关性。方法选取本院接受治疗的135例PCOS患者作为观察组,另选取在本院妇科门诊进行健康体检且各项检查均正常的66例志愿者作为对照组。比较两组的糖脂代谢指标、氧化应激指标和TLR4水平;采用Pearson相关系数法分析观察组血清TLR4与糖脂代谢及氧化应激指标的相关性,并以受试者工作特征(ROC)曲线分析血清TLR4对于PCOS的诊断效能。结果观察组的空腹血糖(PFG)、空腹胰岛素(FINS)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、丙二醛(MDA)和TLR4水平均显著高于对照组,总抗氧化活性(TAA)水平低于对照组(P<0.05)。Pearson相关性分析结果显示,观察组的血清TLR4与FPG、FINS、TG、LDL-C、MDA均呈正相关,与TAA呈负相关(P<0.05)。ROC曲线分析显示,血清TLR4>0.86 ng/mL时对PCOS发生具有一定诊断价值[曲线下面积(AUC)=0.836,95%CI:0.777~0.895],诊断灵敏度为88.15%,特异度为65.15%。结论PCOS患者体内TLR4异常升高,其水平与机体糖脂代谢及氧化应激均存在一定联系,且对PCOS有一定的诊断价值。

宫腔粘连组织中TLR4/NF-κB通路功能改变与细胞因子分泌、胶原代谢特征的相关性

目的:研究宫腔粘连(IUAs)组织中Toll样受体4(TLR4)/核因子κB(NF-κB)通路功能改变与细胞因子分泌、胶原代谢特征的相关性。方法:选择在我院就诊的IUAs患者作为IUAs组,同期因不孕行宫腔镜检查且经病理证实子宫内膜正常的患者作为对照组。测定IUAs组粘连组织及对照组正常子宫内膜组织中TLR4/NF-κB通路分子、胶原代谢基因的表达水平及细胞因子、胶原代谢标志分子的含量。结果:IUAs粘连组织中TLR4、NF-κB、解整合素金属蛋白酶15(ADAM15)、ADAM17、基质金属蛋白酶9(MMP9)、纤溶酶原激活剂抑制剂-1(PAI-1)的mRNA表达量以及转化生长因子-β1(TGF-β1)、Smad2/3、胰岛素样生长因子-1(IGF-1)、IGF-1受体(IGF-1R)、碱性成纤维细胞生长因子(bFGF)、骨膜蛋白成骨细胞特异性因子2(Postn)、I型胶原(Col-I)、肌动蛋白-α(α-SMA)的含量明显高于对照组,尿激酶纤溶酶原激活剂(uPA)的mRNA表达量明显低于对照组;TLR4、NF-κB的mRNA表达量与TGF-β1、Smad2/3、IGF-1、IGF-1R、bFGF、Postn、Col-I、α-SMA、ADAM15、ADAM17、MMP9、PAI-1呈正相关,与uPA呈负相关。结论:IUAs组织TLR4/NF-κB通路的过度激活与细胞因子分泌及胶原代谢异常有关。

TLR4/NF-κB信号通路介导肥胖症患者血清的致炎作用

目的探讨肥胖症患者血清对THP-1单核细胞分泌炎症因子的作用及探讨TLR4/NF-κB信号通路在肥胖症患者血清促炎症作用中的贡献。方法分别用15例单纯性肥胖及伴不同代谢紊乱(血脂紊乱、高血压、高血糖)患者血清孵育THP-1单核细胞株48 h后以及用TLR4单克隆抗体阻断TLR4/NF-κB信号通路后,测定THP-1细胞表面TLR4和细胞内NF-κB p65磷酸化的表达水平及其分泌白细胞介素1β和肿瘤坏死因子α的能力。免疫印迹法检测TLR4蛋白和细胞内NF-κB p65磷酸化蛋白水平;用RT-Q-PCR检测TLR4 mRNA的表达水平;用酶联免疫吸附法检测细胞上清液中白细胞介素1β和肿瘤坏死因子α的水平。结果与正常对照组比较,单纯性肥胖和肥胖症伴代谢紊乱患者血清干预后,THP-1细胞TLR4、NF-κB p65磷酸化蛋白及TLR4 mRNA表达水平以及分泌白细胞介素1β和肿瘤坏死因子α的能力显著升高(P<0.05);THP-1细胞TLR4、NF-κB p65磷酸化蛋白及TLR4mRNA表达水平以及分泌白细胞介素1β和肿瘤坏死因子α的能力随肥胖症代谢紊乱组分的增加而增高,且当TLR4/NF-κB信号通路被阻断后,分泌白细胞介素1β和肿瘤坏死因子α的能力降低。结论单纯性肥胖及肥胖症伴不同代谢紊乱患者的血清可不同程度地诱导THP-1细胞TLR4/NF-κB信号通路的活化,TLR4/NF-κB信号通路在肥胖症患者血清促THP-1分泌炎症因子的作用中起重要作用。

SDF-1/CXCR7生物学轴与肿瘤的研究进展

历来认为趋化因子受体4（chemokine receptor 4,CXCR4）是趋化因子SDF-1的唯一受体,SDF-1/CXCR4生物学轴在肿瘤的发生发展过程中起重要作用,最近研究发现SDF-1存在另一个受体CXCR7,并且SPF-1/CXCR7生物学轴同样对肿瘤的发生发展起着重要的作用.本文就SPF-1/CXCR7生物学轴在肿瘤中的增殖、侵袭、转移以及肿瘤治疗等方面的研究作一综述.

黑素皮质素受体-4基因rs489693位点多态性与慢性精神分裂症患者体质量指数及糖脂代谢水平的关系

目的:探讨慢性精神分裂症患者黑素皮质素受体-4基因(MC4R) rs489693多态性与患者体质量指数(BMI)、糖脂代谢的相关性。方法:135例慢性精神分裂症患者根据DNA测序法检测MC4R基因rs489693位点的多态性分为CC组67例、AC组50例、AA组18例,并测定3组患者体质量、腰围、BMI,血清总胆固醇、三酰甘油、高密度脂蛋白、低密度脂蛋白、血糖、糖化血红蛋白、尿酸及催乳素水平,使用方差分析比较各组间各变量的差异。结果:rs489693基因分布频率均符合Hardy-Weinberg平衡检验(χ2=2. 91,P=0. 09)。CC组、AC组及AA组在体质量(F=4. 041,P=0. 025)、BMI(F=3. 337,P=0. 045)、血清总胆固醇(F=3. 184,P=0. 036) 3项变量差异有统计学意义;事后多重比较发现AA组体质量、BMI、血清总胆固醇3项变量显著高于CC组(P均<0. 05); Logistic回归分析发现BMI与基因型AA有关(OR=22. 071,95%CI:1. 803～270. 22; P=0. 015)。结论:MC4R基因rs489693的A等位基因是慢性精神分裂症患者发生肥胖和高血脂的风险因素。

TLR4与脂代谢的研究进展

Toll样受体4(TLR4)是人类最早发现的一种Toll样受体相关蛋白,普遍分布于多种免疫细胞的膜表面,通过识别脂多糖等外源性配体激活My D88依赖性和非依赖性信号通路,参与炎症反应和免疫应答。近年来,诸多学者对TLR4信号通路介导的炎症反应、脂代谢调控及其相互作用关系进行了系统研究,相关研究表明,TLR4除了在先天性免疫过程中扮演重要角色外,还参与着机体脂代谢的调节过程。文章主要通过阐述TLR4的结构、分布、配体、信号通路及其与脂代谢的相互关系来介绍TLR4的最新研究进展。

Toll样受体4对氧诱导视网膜新生血管发生的促进作用及其机制

背景研究发现，Toll样受体（TLRs）在视网膜新生血管的形成中发挥重要作用，但其作用机制尚不清楚。目的探讨TLR4对小鼠氧诱导视网膜病变（OIR）中新生血管生成的作用及其机制。方法将18只7日龄新生C57BL／6J小鼠以随机数字表法按小鼠窝别分为常氧组、OIR组及0IR＋脂多糖（LPS）-EB组，常氧组小鼠在正常氧环境中饲养，OIR组及OIR＋LPS—EB组小鼠于生后第7天（P7）与母鼠置于体积分数（75±2）％高氧氧箱中饲养5d以诱导OIR模型，于P12在正常氧环境中饲养，OIR＋LPS-EB组P12小鼠腹腔内注射LPS-EB，剂量为1mg／kg，OIR组同法注射PBS。各组摘取P17小鼠眼球于质量分数4％多聚甲醛中固定并行视网膜铺片和Lectin染色，计算视网膜无血管区和新生血管区面积占全视网膜的百分比。各组制备P17小鼠眼后节组织冰冻切片并行免疫荧光检测，计数小鼠5mm视网膜全长活化小胶质细胞数目；采用CDllb和TLR4免疫荧光双标记法检测各组小鼠视网膜小胶质细胞中CDllb、TLR4的表达及其分泌血管内皮生长因子（VEGF）、白细胞介素1p（IL-1β）和肿瘤坏死因子-a（TNF-a）的水平。结果常氧组P17小鼠视网膜血管分布和形态正常，OIR组和OIR＋LPS—EB组P17小鼠视网膜中央均出现无血管区和新生血管簇，OIR＋LPS-EB组小鼠视网膜无血管区面积比为（18．47±1．32）％，新生血管面积比为（3．29±0．85）％，分别大于OIR组的（15．78±1．44）％和（1．77±0．19）％，差异均有统计学意义（t=3．36，P=0．01；t=4．22，P=0．00）。免疫荧光结果显示，常氧组P17小鼠视网膜中活化小胶质细胞数量极少，OIR组和OIR＋LPS—EB组P17小鼠视网膜中活化小胶质细胞荧光强度增强，活化小胶质细胞数量增加，其中OIR＋LPS-EB组小鼠视网膜中小胶质细胞数明显多于OIR组，差异有统计学意义[（95．50±4．77）／5mm与（74．83±4．17）／5mm，t=8．00，P〈0．01]。常氧组小鼠视网膜中TLR4阳性小胶质细胞数量极少，OIR组和OIR＋LPS—EB组小鼠视网膜中TLR4荧光增强，TLR4阳性小胶质细胞数量明显增加，OIR＋LPS—EB组小鼠视网膜中TLR4阳性小胶质细胞数目明显多于OIR组，差异有统计学意义[（49．50±6．38）／5mm与（28．17±6．24）／5mm，t=5．86，P〈0．01）]。常氧组P17小鼠视网膜中CDllb和VEGF／IL-1β／TNF-a共表达的小胶质细胞数量分别为（1．17±0．75）／5mm、0和0，而OIR＋LPS—EB组小鼠视网膜中共表达CDllb和VEGF／IL·1β／TNF—d的小胶质细胞数量多于OIR组，差异有统计学意义[（44．50±8．78）／5mm与（28．50±5．61）／5mm，F=44．07，P〈0．01；（24．10±6．49）／5mm与（16．00±3．46）／5nlm，F=11．31，P〈0．01；（33．83±14．82）／5mm与（23．00±2．83）／5mm，t=19．92，P〈0．01]。结论TLR4可促进OIR小鼠视网膜新生血管的生成，其作用机制可能与TLR4激活视网膜小胶质细胞中相关的下游信号通路，促进血管生长因子及炎性因子的释放有关。

用于多巴胺D_4受体体内研究的3-[4-(4-[^(18)F]氟苯甲基)哌嗪-1-基]甲基-1H-吡咯并[2,3-b]吡啶的合成及生物学评价

目的 7 氮杂吲哚的衍生物L 74 5 ,870是一个新的、高亲和性 (Ki=0 4 3nmol·L-1)的多巴胺D4受体选择性配体 ,我们放化合成了其类似结构的新化合物 3 [4 (4 [18F]氟苯甲基 )哌嗪 1 基 ] 甲基 1H 吡咯并 [2 ,3 b]吡啶 ([18F]C) ,并作了大鼠体内生物学评价。方法 ([18F]C)的放化合成是通过 3 (哌嗪 1 基 )甲基 1H 吡咯并 [2 ,3 b]吡啶和 4 氟[18F]苯甲醛的胺烷基化反应完成 ,放化产率为 9 0 %～12 0 % ,放化纯度大于 98% ,比活度高于 37GBq·mol-1。结果 额叶皮质、海巴和延髓等脑区有较高的放射性摄取率 ,分别为 0 4 3%ID/ g和 0 35 %ID/g ;而纹状体、小脑处放射性摄取率较低。结论 大鼠体内的组织分布和代谢物研究表明 :[18F]C在大鼠脑组织区域有特异性分布 。

慢性情绪应激对高脂饮食大鼠炎症反应和主动脉Toll样受体4表达的影响

目的通过观察慢性情绪应激对高脂饮食大鼠脂代谢、炎症反应和主动脉内皮细胞TLR4表达的影响,探讨慢性情绪应激在动脉粥样硬化病变形成中的作用、机制。方法雄性Wistar大鼠40只,随机分为正常对照组(NC)、无应激组(NS)、生理应激组(PS)和情绪应激组(ES),每组各10只,并制作慢性情绪应激模型。于末次应激结束后采集血标本,全自动生化分析仪检测血清TC、TG、HDL-C、LDL-C和hs-CRP,ELISA法测定血清ox-LDL、放免法检测TNF-α水平;HE染色观察大鼠主动脉形态学变化;免疫组化法(S-P)测定主动脉内皮细胞TLR4表达。结果 (1)ES组大鼠较其他3组出现了明显的脂代谢紊乱和炎症反应。与PS组、NS组、NC组比较,ES组TC、LDL-C、ox-LDL水平明显升高[TC:(5.30±0.69)mmol/L比(3.94±0.42)mmol/L、(3.82±0.48)mmol/L、(2.07±0.26)mmol/L;LDL-C:(1.57±0.22)mmol/L比(1.18±0.13)mmol/L、(1.11±0.11)mmol/L、(0.75±0.11)mmol/L;ox-LDL:(65.18±6.51)μg/dl比(45.65±2.70)μg/dl、(38.35±2.27)μg/dl、(14.99±1.46)μg/dl,均为P<0.01];ES组HDL-C[(0.94±0.14)mmol/L]低于NS组[(1.09±0.14)mmol/L,P<0.05],低于NC组[(1.16±0.18)mmol/L,P<0.01];与PS组、NS组、NC组比较,ES组hs-CRP、TNF-α水平升高[hs-CRP:(1.748±0.082)mg/L比(1.485±0.067)mg/L、(1.381±0.067)mg/L、(0.757±0.069)mg/L;TNF-α:(2.447±0.083)μg/L比(2.189±0.099)μg/L、(2.181±0.085)μg/L、(1.772±0.075)μg/L,均为P<0.01];(2)ES组大鼠主动脉出现早期动脉粥样硬化性改变;(3)ES组大鼠主动脉内皮细胞TLR4表达明显上调,其阳性细胞单位面积平均吸光度值(A)高于PS组、NS组和NC组[(0.334±0.010)比(0.250±0.012)、(0.238±0.015)、(0.082±0.008),均为P<0.01]。结论慢性情绪应激可能在动脉粥样硬化形成的早期,部分通过激活TLR4释放炎性细胞因子引起机体的炎症反应,进而导致动脉粥样硬化病变形成。

模拟代谢性内毒素血症对肝损伤小鼠肝组织TLR4信号通路的影响

目的观察持续4 w的模拟代谢性内毒素血症对小鼠肝脏组织病理和4型Toll样受体(TLR4)信号通路的影响。方法将30只雄性C57BL/6J小鼠随机分为正常组(n=10)和内毒素组(n=10),均予以普通饲料喂养,和高脂组(n=10),予以高脂饲料喂养。内毒素组小鼠同时经腹部皮下植入的微泵注入内毒素300μg·kg-1·d-1,连续4 w,正常组小鼠皮下微泵持续注入生理盐水作为对照。4 w后测定小鼠血清内毒素水平,对肝组织切片行HE染色后进行非酒精性脂肪性肝病评分(NAS),采样Real time PCR法测定小鼠肝组织TLR4及其下游的My D88和TRIF-related adaptor molecule(TRAM)m RNA水平。结果内毒素组小鼠血清内毒素水平(0.62±0.04 EU/ml)显著高于正常组(0.50±0.06 EU/ml,P<0.05)和高脂组(0.49±0.05 EU/ml,P<0.05);内毒素组小鼠肝组织主要表现为单纯性脂肪变性,NAS积分为(2.30±0.49),高脂组小鼠肝组织炎症较明显,NAS积分为(4.20±1.61),显著高于内毒素组(P<0.05);与正常组相比,内毒素组小鼠肝组织TLR4 m RNA水平上调5.12倍(P<0.01),TRAM m RNA水平上调3.46倍(P<0.01),而My D88 m RNA水平与正常组比,无显著差别。结论持续4 w的模拟代谢性内毒素血症可诱导小鼠肝脏单纯性脂肪变性,TLR4 m RNA和TRAM m RNA水平上调,而My D88 m RNA水平无显著变化。