姜黄素对缺氧再给氧大鼠心脏微血管内皮细胞ICAM-1表达的影响

目的：研究姜黄素对缺氧再给氧大鼠心脏微血管内皮细胞粘附分子ICAM－1表达的影响。方法：培养大鼠心脏微血管内皮细胞，建立细胞缺氧再给氧模型，采用酶联免疫吸附试验（ELISA）测定内皮细胞粘附分子ICAM－1的蛋白表达，Northern blot分析测定ICAM－1mRNA的表达。结果：缺氧后内皮细胞ICAM－1的蛋白和mRNA表达明显升高，缺氧再给氧后增高更明显，姜黄素组ICAM－1的蛋白和mRNA的表达较缺氧再给氧组明显下降，呈剂量依赖效应。结论：姜黄素能明显下调缺氧再给氧大鼠心脏微血管内皮细胞ICAM－1的表达，抑制内皮细胞的激活。     

基于网络药理学和分子对接技术探讨生脉注射液抗新型冠状病毒肺炎的作用机制

目的采用网络药理学与分子对接技术探讨生脉注射液的活性成分和治疗新型冠状病毒肺炎(COVID-19)的潜在作用机制。方法利用TCMSP及BATMAN-TCM数据库筛选生脉注射液的活性化合物,通过TCMSP及Targetnet在线数据库预测作用靶点,通过Cytoscape3.7.1构建活性成分-作用靶点网络图;在GeneCards及OMIM数据库中以"coronavirus pneumonia"为关键词搜索冠状病毒肺炎相关疾病靶点,与生脉注射液化合物靶点进行交集筛选出共同靶点作为研究靶点,将共同靶点导入STRING数据库获取数据后在Cytoscape 3.7.1软件中构建蛋白质-蛋白质相互作用网络图;利用R语言进行GO(gene ontology)功能、KEGG(Kyoto encyclopedia of genes and genomes)通路富集分析,预测其作用机制,并构建"成分-靶点-通路"网络图;通过DiscoveryStudio 2.5软件对关键靶点进行分子对接分析。结果生脉注射液筛选得到22个活性化合物,分别为邻苯二甲酸二辛酯、β-谷甾醇、当归酰基戈米辛O、戈米辛A、戈米辛R、五味子丙素、内南五味子酯乙、长南酸、南五味子内酯、香蒲木脂素B、新杜松烷酸A、新杜松烷酸B、新杜松烷酸C、新南五味子木脂宁、五味子内酯A、五味子内酯E、五味子酸、尿苷、薯蓣皂苷元、鸟嘌呤核苷、N-反式阿魏酰酪胺、豆甾醇。相应作用靶点224个,与COVID-19的共同靶点16个,分别为CASP3、CASP8、PTGS2、BCL2、BAX、PRKCA、PTGS1、PIK3CG、F10、NOS3、DPP4、NOS2、TLR9、ACE、ICAM1、PRKCE,关键靶点涉及CASP3、PTGS2、NOS2、NOS3、ICAM1。GO功能富集分析得到生物过程(BP)条目771个,细胞组成(CC)条目11个,分子功能(MF)条目79个。KEGG通路富集分析筛选得到67条(P0.05)信号通路,主要涉及糖尿病并发症AGE-RAGE信号通路、凋亡通路、P53信号通路、小细胞肺癌通路等。分子对接结果显示与关键靶点对接较好的成分有五味子内酯E、豆甾醇、N-反式阿魏酰酪胺。结论生脉注射液中的活性化合物五味子内酯E、豆甾醇、N-反式阿魏酰酪胺等能作用于CASP3、PTGS2、NOS2、NOS3等靶点调节多条信号通路发挥抗炎、免疫调节、抗休克、增加血氧饱和度等作用,从而可能发挥对COVID-19的治疗作用。     

短小棒状杆菌制剂对小白鼠ICAM1/LFA1基因表达的研究

目的采用短小棒状杆菌制剂对小白鼠ICAM1/LFA1基因表达的研究,从而进一步研究短小棒状杆菌制剂对免疫调节和抗肿瘤机理深入研究提出依据。方法本试验采用realtime PCR方法,以GAPDH和-βactin分别作为各组织样品的内源控制基因。结果短小棒状杆菌制剂短小棒状杆菌制剂小白鼠腹腔注射之后,ICAM1基因在脾脏组织中上调,5 h出现高峰,短小棒状杆菌制剂小白鼠腹腔注射之后3-5 h LFA-1基因持续性上调。不同组织的LAF-1基因检测结果显示,小肠和脾脏均表现为高表达,尤以脾脏突出。结论短小棒状杆菌制剂的免疫机理和治疗肿瘤的作用与早期激动免疫细胞的第一信号有直接相关。     

过氧化物酶体增殖物激活受体β/δ激动剂GW0742对硝酸甘油诱导偏头痛大鼠的作用

目的 观察过氧化物酶体增殖物激活受体（peroxisome proliferators-activated receptor β/δ,PPARβ/δ）激动剂GW0742对硝酸甘油诱导偏头痛大鼠的影响.方法 48只（SD大鼠）随机分为模型组、GW0742组和正常对照组.模型组按Tassorelli Cristina法复制大鼠偏头痛模型,GW0742组于造模后30min给药.取三叉神经颈复合体,采用免疫组织化学和Western blot法测定PPARβ/δ及白细胞介素-6（interleukin-6,IL-6）、细胞间粘附分子（intercellular adhesion molecule-1,ICAM-1）、基质金属蛋白酶-9（Matrix metallopeptidase 9,MMP-9）的表达.结果 模型组三叉神经颈复合体中PPARβ/δ的表达明显高于对照组,差异有统计学意义,同时模型组大鼠IL-6、ICAM-1、MMP-9高于对照组,给予GW0742大鼠IL-6、ICAM-1、MMP-9表达明显下降.结论 PPARβ/δ在偏头痛时高度表达,PPARβ/δ激动剂对偏头痛有保护作用.     

免疫性间质性肾炎动物模型肾组织炎症细胞浸润与细胞间粘附分子-1的表达

目的 探讨自身免疫性间质性肾炎的发病机理。方法 用正常小牛肾小管基底膜(bTBM)制造抗TBM型BN大鼠间质肾炎模型,并用单克隆抗体免疫组化ABC法、间接免疫荧光技术,检测炎症细胞及细胞间粘附分子-1(ICAM-1)等。结果 用bTBM免疫后第9天～3周,均可见肾间质有炎症细胞浸润及肾小管上皮细胞有ICAM-1强烈表达,与病理损害程度有同步现象。结论 在免疫性间质性肾炎发生发展中,由ICAM-1介入的细胞免疫起重要作用,但不能除外体液免疫的参与。     

Intercellular adhesion molecule-3 is the predominant co-stimulatory ligand for leukocyte function antigen-1 on human blood dendritic cells

Dendritic cells (DC) are potent stimulators of primary T lymphocyte responses to foreign antigen. The initial DC-T lymphocyte interaction involves the binding of the adhesion molecule leukocyte function antigen-1 (LFA-1; CD11a/CD18) on the T lymphocyte to an intercellular adhesion molecule (ICAM) on the DC. Although blood and tonsil DC express ICAM-1 (CD54) and ICAM-2 (CD102) on their surface, anti-ICAM-1 and anti-ICAM-2 monoclonal antibodies (mAb) have little inhibitory activity on the DC-stimulated mixed leukocyte reaction (MLR). We therefore examined the expression of the more recently identified LFA-1 ligand, ICAM-3 (CD50), in comparison to ICAM-1 and ICAM-2 on blood DC and sought a functional role for ICAM-3 in DC-mediated T lymphocyte responses. Resting blood DC expressed significantly more ICAM-3 than ICAM-1 or ICAM-2 as assessed by flow cytometry. Treatment of resting DC with interferon-γ led to increased expression of ICAM-1; however, ICAM-2 and ICAM-3 levels remained relatively constant. Solid-phase recombinant chimeric molecules ICAM-1-, ICAM-2- and ICAM-3-Fc were able to co-stimulate CD4⁺ T lymphocyte proliferation in conjunction with suboptimal solid-phase CD3 mAb 64.1. However, the anti-ICAM-3 mAb CAL 3.10 inhibited a DC-stimulated MLR to a greater extent than anti-ICAM-1 or anti-ICAM-2 reagents and appeared to act by blocking the DC ICAM-3- T lymphocyte LFA-1 interaction. As ICAM-3 is the predominant LFA-1 ligand on resting blood DC, we postulate that DC may utilize ICAM-3 for initial DC-T lymphocyte interactions, and that ICAM-1, which is up-regulated upon DC activation, and/or ICAM-2, may contribute to DC migration or later phases of the T lymphocyte activation process.     

Anti-atherosclerotic effects of Polygonum aviculare L. ethanol extract in ApoE knock-out mice fed a Western diet mediated via the MAPK pathway

Ethnopharmacological relevance

Polygonum aviculare L. has been used in traditional Korean medicine to treat obesity and symptoms associated with hypertension. The effectiveness or mechanism of Polygonum aviculare L. ethanol extract (PAE) on atherosclerosis disease has not been examined experimentally. This study investigated the protective effect of PAE in atherosclerotic mice.

Materials and methods

ApoE KO mice were fed a Western diet (WD) alone or with PAE or a statin for 12 weeks, followed by analysis of bodyweight, serum lipid levels, and blood pressure. Staining of the aorta and adipose tissue, expression levels of adhesion molecules, and the MAPK pathway were also examined. Cell viability, NF-κB activity, and protein levels of adhesion molecules were assessed in vitro.

Results

ApoE KO mice fed PAE (50 and 100 mg/kg) or statin (10 mg/kg) gained less body weight, and has less adipose tissue and lower serum lipid levels and blood pressures than the WD group. Aorta ICAM-1, VCAM-1, and NF-κB levels were decreased by PAE in a dose-dependent manner, consistent with the in vitro observations. PAE and statin decreased atherosclerotic plaque and adipocyte size versus the WD group. Furthermore, PAE decreased phosphorylation of MAPK pathway components in the aorta of PAE-treated mice, suggesting that PAE's anti-atherosclerotic effects are mediated via a MAPK pathway-dependent mechanism.

Conclusions

PAE may protect against the development of atherosclerotic disease. The beneficial effects are associated with lowering bodyweight, serum lipids, blood pressure, adhesion molecular protein levels, atherosclerotic plaque, and adipocyte size, involving the MAPK pathway.