睾酮对脂多糖损伤的血管内皮细胞功能的影响

目的探讨睾酮对脂多糖损害的血管内皮细胞功能的影响。方法将培养3代的人脐静脉内皮细胞分为7组,对照组细胞不处理;脂多糖组加脂多糖10 mg/L刺激;5个不同浓度睾酮组,在脂多糖刺激同时,分别加入3×10~(10)、3×10~(-9)、3×10~(-8)、3×10~(-6),3×10~(-5)mol/L浓度睾酮(依次为A、B、C、D、E组)。硝酸还原酶法检测NO含量,RT-PCR检测内皮型一氧化氮合酶(eNOS)与单核细胞趋化因子-1(MCP-1)mRNA水平;ELISA法检测MCP-1蛋白。结果与脂多糖组比较,B组、C组及D组NO含量和eNOS mRNA均显著增加(P<0.05,P<0.01);与对照组MCP-1蛋白(374.16±10.2)ng/L比较,A组明显增加[(424.50±11.3)ng/L,P<0.05];随睾酮浓度增加,MCP-1蛋白逐渐减少,E组MCP-1蛋白显著减少[(292.29±12.6)ng/L,P<0.01]。与对照组MCP-1mRNA比较,B组、D组、E组明显减低(P<0.05.P<0.01)。结论生理浓度睾酮通过促进NO合成,减少MCP-1表达、改善脂多糖损害的血管内皮细胞功能而发挥抗动脉粥样硬化作用。     

Pulmonary delivery of siRNA against acute lung injury/acute respiratory distress syndrome

The use of small interfering RNAs (siRNAs) has been under investigation for the treatment of several unmet medical needs, including acute lung injury/acute respiratory distress syndrome (ALI/ARDS) wherein siRNA may be implemented to modify the expression of pro-inflammatory cytokines and chemokines at the mRNA level. The properties such as clear anatomy, accessibility, and relatively low enzyme activity make the lung a good target for local siRNA therapy. However, the translation of siRNA is restricted by the inefficient delivery of siRNA therapeutics to the target cells due to the properties of naked siRNA. Thus, this review will focus on the various delivery systems that can be used and the different barriers that need to be surmounted for the development of stable inhalable siRNA formulations for human use before siRNA therapeutics for ALI/ARDS become available in the clinic.     

LPSp对狂犬病疫苗佐剂效应及安全性研究

目的研究成团泛菌脂多糖(LPSp)作为狂犬病疫苗佐剂的可行性。方法将Balb/c小鼠32只随机分为空白对照组(注射生理盐水)、LPSp对照组(注射LPSp)、狂犬病病毒蛋白组(注射狂犬病病毒蛋白)、LPSp实验组(注射LPSp和狂犬病病毒蛋白),分别在第0、7、15、21天经背部皮下免疫接种共4次。在免疫接种后第7、14、21、30、45、60天采集小鼠眼内眦静脉血,分离血清,末次采血后处死小鼠。用酶联免疫吸附试验(ELISA)检测血清抗狂犬病病毒IgG,并进行血液学、血液生化、脏器指数等检查。结果免疫接种4次后,LPSp能明显增强狂犬病病毒蛋白免疫小鼠产生IgG的能力(P<0.05);实验期间各组小鼠未发现明显异常表现,体质量、血液学、血液生化和脏器指数等各组间未见明显差异(P>0.05)。结论 LPSp对狂犬病病毒蛋白具有较好佐剂效应;在本实验条件下,LPSp对小鼠安全性好。     

经侧脑室注射脂多糖诱导大鼠黑质部位小胶质细胞激活及多巴胺能神经元损伤的研究

目的观察经脑室注射脂多糖(LPS)后大鼠的黑质部小胶质细胞激活及多巴胺(DA)能神经元的变化,探讨脑内炎性反应在黑质DA能神经元慢性变性过程中的作用。方法健康雄性SD大鼠30只,随机分为生理盐水(NS)对照组和LPS组,分别向大鼠右侧脑室注射20μL NS或50μg LPS,40周后用免疫组织化学方法检测大鼠黑质小胶质细胞是否激活、激活的程度(OX-42及OX-6抗体水平),以及酪氨酸羟化酶(TH)阳性神经元的形态和数量。以Fluoro-Jade B(FJB)染色法检测黑质部位神经元变性情况。结果 (1)NS对照组大鼠黑质部位OX-42阳性小胶质细胞呈静息状态,染色浅。LPS组大鼠黑质部OX-42阳性小胶质细胞呈部分激活状态,染色深。两组大鼠黑质部位均未发现OX-6阳性小胶质细胞。(2)NS对照组大鼠黑质部位有大量深染的TH阳性神经元。LPS组大鼠黑质部位TH阳性染色神经元数目(99.11±20.31)比NS对照组(189.52±12.12)减少47.7%(P<0.01)。(3)两组大鼠黑质部位均未见FJB阳性染色神经元。结论经侧脑室单次注射LPS可能造成大鼠黑质部位小胶质细胞长期慢性激活及DA能神经元慢性迟发性功能性损伤。     

Stress Response and Apoptosis in Pro- and Antiinflammatory Macrophages

We showed that stress response and apoptosis in macrophages depend on the phenotype of their secretory activity and specific biological and physical characteristics of the factor inducing stress-response or apoptosis.     

“Latent” healthy human serum autoantibodies cross-reacting with DNA and bacterial lipopolysaccharides

Department of Radiation Biochemistry, Research Institute of Medical Radiology, Academy of Medical Sciences of the USSR, Obninsk. (Presented by Academician of the Academy of Medical Sciences of the USSR V. A. Nasonova.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 111, No. 5, pp. 516–518, May, 1991.     

3-O-C12-HSL通过抑制lncRNA CD48-AS和CD48的表达而阻碍Mo-DCs成熟

目的 筛选并阐明长链非编码RNA（lncRNA）CD48-AS与其反义互补分子CD48 mRNA在N-3-氧代十二 烷酰-L-同型丝氨酸内酯（3-O-C12-HSL）阻碍人单核细胞诱导的树突状细胞（Mo-DCs）成熟过程中发挥作用的机制。 方法 从健康人外周血中分离Mo-DCs,将未成熟的Mo-DCs细胞分为阴性对照组（0.1% DMSO）、脂多糖（LPS）阳性 对照组（100 μg/L的LPS）和实验组（100 μg/L LPS+40 μmol/L 3-O-C12-HSL）进行lncRNA芯片检测。筛选lncRNA表 达谱中差异表达 5 倍以上的自然反义 lncRNA 进行聚类分析,从中筛选差异具有统计学意义的自然反义 lncRNA CD48-AS。未成熟的 Mo-DCs 分为阴性对照组、LPS 阳性对照组以及 LPS+C5、C10、C25 实验组（分别加入 5、10、 25 μmol/L的3-O-C12-HSL）。利用荧光定量PCR检测lncRNA CD48-AS和反义分子CD48 mRNA在实验体系中的表 达水平;通过生物信息学分析lncRNA CD48-AS与CD48反义互补区及其蛋白编码功能;通过核糖核酸酶保护实验 （RPA）验证lncRNA CD48-AS是否与CD48形成二聚体,从而通过影响靶CD48的表达来发挥调控作用。最后检测 lncRNA CD48-AS在细胞内的定位。结果 3-O-C12-HSL处理Mo-DCs后lncRNA表达谱出现了特异性改变。3-OC 12-HSL可下调由LPS诱导的Mo-DCs中lncRNA CD48-AS及其反义靶分子CD48的表达。生物信息学分析lncRNA CD48-AS 不具备蛋白编码功能,为非编码 RNA;lncRNA CD48-AS 与 CD48 形成 RNA 二聚体从而减少 RNA 酶对 CD48 mRNA的降解,使得CD48 mRNA表达增加;lncRNA CD48-AS在Mo-DCs中主要定位在细胞核中,细胞质中表 达量较少。结论 3-O-C12-HSL可通过下调lncRNA CD48-AS,进而影响CD48的表达来阻碍Mo-DCs的成熟。     

Therapeutic Strategies to Protect the Central Nervous System against Shiga Toxin from Enterohemorrhagic Escherichia coli

Infection with Shiga toxin-producing Escherichia coli (STEC) may cause hemorrhagic colitis, hemolytic uremic syndrome (HUS) and encephalopathy. The mortality rate derived from HUS adds up to 5% of the cases, and up to 40% when the central nervous system (CNS) is involved. In addition to the well-known deleterious effect of Stx, the gram-negative STEC releases lipopolysaccharides (LPS) and may induce a variety of inflammatory responses when released in the gut. Common clinical signs of severe CNS injury include sensorimotor, cognitive, emotional and/or autonomic alterations. In the last few years, a number of drugs have been experimentally employed to establish the pathogenesis of, prevent or treat CNS injury by STEC. The strategies in these approaches focus on: 1) inhibition of Stx production and release by STEC, 2) inhibition of Stx bloodstream transport, 3) inhibition of Stx entry into the CNS parenchyma, 4) blockade of deleterious Stx action in neural cells, and 5) inhibition of immune system activation and CNS inflammation. Fast diagnosis of STEC infection, as well as the establishment of early CNS biomarkers of damage, may be determinants of adequate neuropharmacological treatment in time.     

Tanshinone IIA prevents LPS-induced inflammatory responses in mice via inactivation of succinate dehydrogenase in macrophages

Metabolic reprogramming is associated with NLRP3 inflammasome activation in activated macrophages, contributing to inflammatory responses. Tanshinone IIA (Tan-IIA) is a major constituent from Salvia miltiorrhiza Bunge, which exhibits anti-inflammatory activity. In this study, we investigated the effects of Tan-IIA on inflammation in macrophages in focus on its regulation of metabolism and redox state. In lipopolysaccharides (LPS)-stimulated mouse bone marrow-derived macrophages (BMDMs), Tan-IIA (10 μM) significantly decreased succinate-boosted IL-1β and IL-6 production, accompanied by upregulation of IL-1RA and IL-10 release via inhibiting succinate dehydrogenase (SDH). Tan-IIA concentration dependently inhibited SDH activity with an estimated IC₅₀ of 4.47 μM in LPS-activated BMDMs. Tan-IIA decreased succinate accumulation, suppressed mitochondrial reactive oxygen species production, thus preventing hypoxia-inducible factor-1α (HIF-1α) induction. Consequently, Tan-IIA reduced glycolysis and protected the activity of Sirtuin2 (Sirt2), an NAD⁺-dependent protein deacetylase, by raising the ratio of NAD⁺/NADH in activated macrophages. The acetylation of α-tubulin was required for the assembly of NLRP3 inflammasome; Tan-IIA increased the binding of Sirt2 to α-tubulin, and thus reduced the acetylation of α-tubulin, thus impairing this process. Sirt2 knockdown or application of Sirt2 inhibitor AGK-2 (10 μM) neutralized the effects of Tan-IIA, suggesting that Tan-IIA inactivated NLRP3 inflammasome in a manner dependent on Sirt2 regulation. The anti-inflammatory effects of Tan-IIA were observed in mice subjected to LPS challenge: pre-administration of Tan-IIA (20 mg/kg, ip) significantly attenuated LPS-induced acute inflammatory responses, characterized by elevated IL-1β but reduced IL-10 levels in serum. The peritoneal macrophages isolated from the mice displayed similar metabolic regulation. In conclusion, Tan-IIA reduces HIF-1α induction via SDH inactivation, and preserves Sirt2 activity via downregulation of glycolysis, contributing to suppression of NLRP3 inflammasome activation. This study provides a new insight into the anti-inflammatory action of Tan-IIA from the respect of metabolic and redox regulation.