自体血清与胎牛血清培养兔关节软骨细胞的生物学特性

背景：目前培养软骨细胞多使用胎牛血清。但近年异种血清培养组织向临床应用的安全性受到质疑,因此自体血清培养越来越受到重视。 目的：比较体积分数10%兔自体血清与体积分数10%胎牛血清培养兔关节软骨细胞生物学特性的差异。 方法：兔自体血清培养液制备后,分离培养兔关节软骨细胞,分别在体积分数10%自体血清和体积分数10%胎牛血清中进行单层传代培养至1,3,5代,采用光镜观察细胞形态变化,绘制生长曲线评估细胞增殖速度,观察甲苯胺蓝染色、Ⅰ,Ⅱ型胶原免疫组织化学染色结果,以流式细胞仪分析细胞Ⅰ,Ⅱ型胶原以及CD26,CD44的表达变化。 结果与结论：①在细胞形态上自体血清和胎牛血清培养的软骨细胞差异不大。②自体血清培养的软骨细胞较胎牛血清培养的软骨细胞生长速度更快。③甲苯胺蓝染色结果示,无论是自体血清还是胎牛血清所培养的细胞,染色随代龄的增加逐渐变浅,细胞传至第5代时两组几乎均无异染。对于1代和3代细胞而言,自体血清培养的软骨细胞较胎牛血清所培养的软骨细胞较为深染。④Ⅰ型胶原的表达随代数的增加而增加,而Ⅱ型胶原的表达则随代数的增加而减少。在3代时自体血清培养的软骨细胞Ⅰ型胶原的表达水平低于胎牛血清培养的软骨细胞(P < 0.05);在1代和3代时自体血清培养的软骨细胞Ⅱ型胶原的表达水平高于胎牛血清培养的软骨细胞(P < 0.05)。⑤CD26表达呈现先升高后降低的趋势,而CD44的表达不随传代数的增加而改变。提示同异体体积分数10%的胎牛血清相比,体积分数10%的自体血清培养的兔软骨细胞生长速度快,且在大部分指标上可以较好地保持细胞表型。     

人肾小管上皮细胞血清饥饿法同步化方法的研究

目的：探讨采用血清饥饿法建立将人肾小管上皮细胞（HKC）的同步化方法。方法：将HKC分为6个不同低血清浓度组及对照组，分别采用胎牛血清浓度为0、1、2、3、4、5和100mL／L的DMEM培养液培养24h。根据流式细胞术结果选择的最佳同步化HKC的胎牛血清浓度。将HKC的同步化处理时间分别延长为24h、48h和72h，选择最佳同步化时间。用MTT法绘制生长曲线观察血清饥饿处理对HKC生长的影响。采用免疫细胞化学方法验证血清饥饿同步化处理HKC的效果。结果：选择无血清（0mL／L）和1mL／L为HKC最佳同步化的胎牛血清浓度，选择48h为HKC同步化最佳时间。细胞生长曲线表明采用1mL／L胎牛血清的DMEM培养液培养48h后恢复正常培养的HKC细胞，其生长曲线更接近于正常培养的细胞。Brdu进一步证明，用1mL／L胎牛血清的DMEM培养液培养48h可以使90％以上的HKC细胞处于G0／G1期。结论：用1mL／L胎牛血清的DMEM培养液培养48h可获得90％以上的G0／G1期HKC。     

梯度血清递减体外培养人脐带间充质干细胞

背景：人脐带间充质干细胞的扩增与培养条件密切相关,而含体积分数10%~20%胎牛血清的培养基可促进细胞生长。 目的：建立梯度血清递减扩增培养脐带间充质干细胞的培养技术。 方法：采用胶原酶Ⅱ消化分离获得脐带间充质干细胞悬液,并通过贴壁培养进行纯化,细胞贴壁后采用梯度血清递减方法,即第1代80%含血清培养基,20%无血清培养基;第2代50%含血清培养基,50%无血清培养基;第3代20%含血清培养基,80%无血清培养基;第4代100%无血清培养基。另一种传代中始终采用含体积分数10%胎牛血清的α-MEM完全培养液。用流式细胞仪检测细胞的表面标记,进行成骨诱导试验,同时与经典的含10%胎牛血清的α-MEM培养体系进行比较。 结果与结论：梯度血清递减培养法与经典α-MEM培养体系所获得的细胞在扩增能力、细胞形态、免疫表型等方面相似。细胞在两种培养体系中均能保持良好的分化潜能。但梯度血清浓度法培养间充质干细胞可使用更少量胎牛血清,提高临床应用安全性。     

新生牛血清促细胞生长效应的研究

为探索我国新生牛血清的质量标准,本文着重在新生牛血清促进人外周血淋巴细胞和羊水细胞生长效应方面与美国GIBCO胎牛血清进行了比较实验研究。取30个批号的混合新生牛血清,每批号分别配制RPMI1640和HAMF—10两种培养基。通过外周血淋巴细胞培养,测定细胞分裂指数和分裂指数比;通过羊水细胞培养,测定细胞附着平和相对附着率。结合其他物理、化学、生物等项检测标准,最后确定新生牛血清等级。     

角质细胞无血清培养基促进大鼠毛囊干细胞的增殖

背景：以往研究培养毛囊干细胞多使用DMEM/F12+体积分数10%胎牛血清培养基,而进年来开发出的角质细胞无血清培养基也可应用于毛囊干细胞的培养。 目的：观察3种不同培养基对大鼠毛囊干细胞增殖情况及干细胞纯度的影响。 方法：取大鼠触须部皮肤组织,体式显微镜下分离出毛囊组织,中性蛋白酶Ⅱ与胰蛋白酶和乙二胺四乙酸混合液“两步酶法”消化。所得细胞悬液按细胞数平均分为3份,分别使用角质细胞无血清培养基、DMEM/F12+体积分数10%胎牛血清及角质细胞无血清培养基+体积分数10%胎牛血清共3种培养基培养,Ⅳ型胶原差速贴壁法筛选毛囊干细胞,进行传代培养。 结果与结论：培养毛囊干细胞传至第3代,锥虫蓝染色计数法检测结果显示,此3组间细胞活率差异无显著性意义(P > 0.05)。CCK8比色法检测细胞生长曲线显示,培养前2 d,此3组细胞生长均较缓慢;培养4 d,细胞生长进入对数生长期,3种培养基培养的细胞增殖活性：角质细胞无血清培养基+10%胎牛血清组>DMEM/F12+10%胎牛血清>角质细胞无血清培养基(P < 0.05)。流式细胞仪检测显示,角质细胞无血清培养基组CD34的表达高于角质细胞无血清培养基+10%胎牛血清组(P < 0.05)。DMEM/F12+10%胎牛血清组中CD34、β1-整合素(CD29)及CK15标记物的表达低于其他2组(P < 0.05)。结果表明,角质细胞无血清培养基较DMEM/F12+体积分数10%胎牛血清能培养出纯度更高的毛囊干细胞,并且在此培养基培养的基础上加入血清,能够促进毛囊干细胞的增殖。     

同种异体与自体血清对骨髓基质干细胞的增殖效应

背景：传统的细胞培养方法大多采用胎牛血清作为营养支持,其存在传播疾病的风险和免疫排斥反应。寻找胎牛血清替代物并建立规范、安全、高效的成人骨髓基质干细胞体外培养体系刚刚起步,有待进一步研究。 目的：探讨AB血清、自体血清替代胎牛血清体外培养人骨髓基质干细胞的效果。 方法：从成人骨髓穿刺液中分离人骨髓基质干细胞,分别用含体积分数10%AB血清、自体血清和胎牛血清的基础培养基培养人骨髓基质干细胞,取第3代细胞进行相关指标检测。 结果与结论：相差显微镜下见AB血清组细胞最先达到汇合;流式细胞仪检测细胞表面抗原,各组均符合骨髓干细胞表型,纯化程度良好;Alamar Blue法显示AB血清组平均荧光强度最高,促增殖效率最高;Annexin V-FITC法显示各组细胞凋亡率均在5%以下,生长状态良好;碱性磷酸酶染色、钙结节和油红O染色结果显示,各组细胞均保持了成骨、成脂分化能力。AB血清对人骨髓基质干细胞的增殖作用较胎牛血清、自体血清明显增强。AB血清有望替代胎牛血清建立符合骨组织工程临床应用要求的体外成人骨髓基质干细胞培养体系。 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

适合人肌母细胞的无血清培养基

背景：肌母细胞体外培养大多使用添加胎牛血清培养基,存在很多不足,开发无血清培养基,更加稳定、安全、经济,有广泛的应用前景。 目的：初步探索适合人肌母细胞培养的无血清培养基。 方法：配制含胎牛血清培养基 (DMEM/F12 1∶1添加胰岛素样生长因子1、碱性成纤维细胞生长因子和体积分数20%胎牛血清),间充质干细胞无血清培养基组(无血清培养基、2%血清替代物、2 mmol/L L-谷氨酰胺,人脐带间充质干细胞,细胞纯度大于99%,UltraCULTURETM添加血清替代物,谷氨酰胺)。自制无血清培养基组(GibcoTM添加胰岛素样生长因子1,碱性成纤维细胞生长因子,谷氨酰胺)。3种培养基分别对人肌母细胞分别进行培养,观察肌母细胞的形态、免疫组织化学的鉴定,计算克隆的形成率,绘制细胞生长的曲线,MTT检测肌母细胞的活性,比较3种培养基培养肌母细胞增殖、分化的差异。 结果与结论：各组细胞形态上无明显差异;各组免疫组织化学鉴定肌母细胞纯度均达99%;含胎牛血清培养基组和自制无血清培养基组克隆形成率显著高于间充质干细胞无血清培养基组(P < 0.01),含胎牛血清培养基组克隆形成率显著高于自制无血清培养基组(P < 0.01);含胎牛血清培养基组与自制无血清培养基组细胞数远高于间充质干细胞无血清培养基组;自制无血清培养基组细胞活性显著高于含胎牛血清培养基组和间充质干细胞无血清培养基组(P < 0.01)。自制无血清培养基组可有效用于肌母细胞的培养,在促进肌母细胞早期贴壁、增殖上还需进一步优化。  中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程      

藻酸钙凝胶固定法能增进无血清培养基中杂交瘤产生抗体的稳定性

虽然在杂交瘤细胞培养中加入胎牛血清能促进细胞生长,但胎牛血清价格昂贵。其组分尚未完全弄清,再加之其质量不稳定,它还常常是微生物污染的来源,血清带入到培养液中的蛋白质,使得抗体的纯化困难。这些都是使用胎牛血清的不利因素。人们进行了大量的研究以期减少或停止使用血清。现已有好几种无血清的杂交瘤细胞培养基应市。然而,研究发现,杂交瘤细胞在无血清培养基中长期培养会丧     

严重烧伤大鼠血清对脂肪间质干细胞生物学特性的影响

目的 探讨严重烧伤大鼠血清对体外培养的大鼠脂肪间充质干细胞(ADSCs)生物学特性的影响.方法 取雄性SD大鼠双侧腹股沟脂肪组织,采用胶原酶消化法、分离纯化大鼠ADSCs,取第3代细胞,采用流式细胞仪检测细胞表面标志物,行成脂成骨诱导分化鉴定;ADSCs传代后按随机数字表法分为10％胎牛血清组(正常培养组)、10％正常大鼠血清组,10％烧伤大鼠血清组,采用噻唑蓝(MTT)法测定各组细胞的生长曲线.通过流式细胞仪检测各组细胞的生长周期、Real-time PCR法检测各组细胞血管内皮生长因子(VEGF)、凋亡相关因子Caspase-3的表达情况.结果 分离培养的ADSCs传至第3代时形态规则,经流式细胞仪检测,所培养的ADSCs均表达CD29、CD90、CD105,阳性率分别为88.6％、99.7％、92.8％,而CD31、CD34、CD45阳性率分别为4.4％、4.7％、3.3％;经诱导培养后可向成骨成脂分化.MTT法检测结果显示:10％烧伤大鼠血清组细胞增殖较快,各时间点吸光度值(A值)明显高于10％胎牛血清组及10％正常大鼠血清组.流式细胞仪检测结果显示:培养1周后10％烧伤大鼠血清组、10％正常大鼠血清组、10％胎牛血清组ADSCs G1期细胞比例分别为:(72.20±5.13)％、(83.50±4.74)％、(90.20±6.37)％;S期细胞比例分别为:(21.40±2.84)％、(12.50±1.96)％和(8.60±1.31)％,10％烧伤大鼠血清组ADSCs G1期细胞比例显著低于10％正常大鼠血清组、10％胎牛血清组(t=2.39、4.86;P ＜0.05);10％烧伤大鼠血清组ADSCs S期细胞比例显著高于10％正常大鼠血清组、10％胎牛血清组(=5.54,7.93;P＜0.01).Real-Time PCR检测结果显示:10％烧伤大鼠血清组VEGF的表达明显上调,Caspase-3表达显著下调与10％正常大鼠血清组、10％胎牛血清组相比差异均有统计学意义(P＜0.05).结论 10％烧伤大鼠血清可明显促进ADSCs的增殖、抑制细胞凋亡发生、促进ADSCs的分泌功能,严重烧伤后可望移植ADSCs用?     

人胚胎纹状体来源神经干细胞的体外培养

背景：目前神经干细胞多由动物获得,不适合人类临床移植治疗。 目的：探索体外环境下人胚胎纹状体来源神经干细胞的培养方法,同时观察其生物学特性。 方法：取经水囊引产的孕8-16周人胚胎纹状体,体外用无血清DMEM培养基进行培养,待细胞形成神经球后进行传代,并应用含体积分数10%胎牛血清的DMEM/ F12培养液进行诱导分化。 结果与结论：体外培养的人胚胎纹状体来源神经干细胞生长迅速,表达神经干细胞标志物nestin。克隆形成实验显示细胞克隆形成率为6.0%-7.0%;BrdU掺入实验显示细胞增殖率为37.9%。免疫荧光染色显示经诱导分化的细胞表达神经元标志物Ⅲ型β微管蛋白、星形胶质细胞标志物胶质纤维酸性蛋白及神经干细胞标志物nestin,但不表达少突胶质细胞标志物髓鞘碱性蛋白。可见人胚胎纹状体来源神经干细胞在体外无血清条件下可保持其生物学特点,具有自我更新能力,经胎牛血清诱导后可向神经元及星形胶质细胞分化。     

Cell-cycle perturbation in Sf9 cells infected with Autographa californica nucleopolyhedrovirus.

Flow cytometry analysis of the cell-cycle progression was performed in Sf9 cells infected with Autographa californica nucleopolyhedrovirus (AcNPV) in the cultures partially synchronized by aphidicolin exposure and deprivation. Cells infected with AcNPV during the G1 phase progressed and were arrested in the S phase in the 4 h following the infection, whereas cells infected during the S phase did not progress past the S phase. Cells infected during the G2/M phase remained in the G2/M phase without mitosis during a period of 10 h. Such cell-cycle arrest was also observed in the cells infected with ts8, a temperature-sensitive mutant of AcNPV that is defective in both genomic DNA synthesis and late gene expression. Cells with >4 N DNA content accumulated in the cultures infected with wild-type AcNPV, whereas no such cells appeared in the cultures infected with ts8, suggesting that viral origin of the DNA overaccumulated in the cells with >4 N DNA content. This was confirmed by the slot blot hybridization experiments, which showed that viral DNA, but not cellular DNA, increased strikingly in Sf9 cells during the infection with AcNPV. These results indicate that AcNPV targets at least two different checkpoints to prevent normal cell-cycle progression of Sf9 cells and that neither viral DNA replication nor expression of viral late genes is a necessary prerequisite for such AcNPV-induced cell-cycle arrest. It is suggested that the cell-cycle arrest in AcNPV-infected Sf9 cells is an event triggered early in infection by specific interaction of viral gene products with cellular components that regulate cell-cycle progression.     

Oxidative stress and DNA damage induced by spinosad exposure in Spodoptera frugiperda Sf9 cells

Spinosad, a neurotoxic insecticide, is widely used for crop protection. In order to elucidate the effects of spinosad on oxidative stress and genotoxicity in Sf9 cells, the levels of lipid peroxidation, the activity of antioxidative enzymes, and DNA damage were measured. The results showed that spinosad caused a time-dependent increase in the formation of malondialdehyde and decrease in the activity of superoxide dismutase and catalase. Further studies confirmed that spinosad induced 8-oxoguanine accumulation in Sf9 cells, which is accompanied by increased expression of DNA repair enzymes (OGG1 and MTH1). The neutral comet assay revealed that spinosad induced significant time-related increases of DNA double-strand breaks in Sf9 cells. Our results indicate that spinosad effectively induced oxidative stress and DNA damage in Sf9 cells.     

Clusters of Cl− channels in CFTR-expressing Sf9 cells switch spontaneously between slow and fast gating modes

The Sf9 insect Spodoptora frugiperda cell line was used for heterologous expression of the cloned human cystic fibrosis transmembrane conductance regulator (CFTR) cDNA, or the cloned β-galactosidase gene, using the baculovirus Autographa califonica as the infection vector. Using application of the patch-clamp technique, evidence for functional expression of CFTR was obtained according to the following three criteria. Firstly, whole-cell currents recorded 2 days after infection with CFTR revealed a statistically significant increase of membrane conductance, ≈25 times above that of mock-infected control cells, with the reversal potential of the major current component being governed by the chloride equilibrium potential (E _Cl). Secondly, in contrast to uninfected cells and cells infected with β-galactosidase, the membrane conductance to chloride of CFTR-injected cells was stimulated by cytosolic adenosine 3′,5′-cyclic monophosphate (cAMP), which was raised by exposing the cells to 10 μM forskolin. Thirdly, recordings of currents through single channels in excised outside-out membrane patches of CFTR-infected cells revealed channels which were clearly different from the native insect chloride channel. Excised outside-out patches of CFTR-infected and forskolin-stimulated cells exhibited wave-like gating kinetics of well-resolved current transitions. All-point Gaussian distributions revealed contributions from several (five to nine) identical channels. Such channels, in excised outside-out patches, studied with a pipette [Cl⁻] = 40 mM and a bath [Cl⁻] = 150 mM, rectified the current in agreement with simple electrodiffusion and with a single-channel Goldman-Hodgkin-Katz permeability, P _Cl = 1.34⋅10⁻¹⁴ ± 0.23⋅ 10⁻¹⁴cm³/s (n = 5), corresponding to a physiological single-channel conductance of 2.8 ± 0.5 pS (V _M = E _Cl) and a limiting conductance, γ_150/150, = 7.7 ± 1.3 pS ([Cl⁻]_Bath = [Cl⁻]_Cell = 150 mM). Currents recorded from multichannel excised outside-out patches could shift from the above mode of resolvable unitary conductance transitions to one which was too fast to reveal the dwell-times of closed and open states. During periods characterized by noisy currents, the variance (σ²) of current fluctuations about their stationary mean value depicted a U-shaped function of membrane potential, with a minimum value at a pipette potential where the chloride current was shown to be zero. Thus, it can be concluded that the current fluctuations are caused by fast gating of channels specific for chloride ions. Switching back and forth between the two gating modes of clusters of chloride channels occurred from moment to moment in excised patches when the membrane potential was held at a constant value indicating cooperative gating as a result of interaction between neighbouring chloride channels. Received: 6 November 1995/Received after revision: 1 February 1996/Accepted: 23 February 1996     

小鼠白细胞介素6的cDNA克隆及其在昆虫细胞中的表达

本文采用ＲＴ－ＰＣＲ技术从ＬＰＳ刺激后的小鼠腹腔巨噬细胞，扩增小鼠ＩＬ－６ｃＤＮＡ，并克隆构建ｐＧＥＭ－３Ｚｆ（＋）ＩＬ－６质粒，继将小鼠所得ｃＤＮＡ克隆到ｐＶＬ１３９２载体上，利用杆状病毒ＡｃＮＰＶ表达系统在Ｓｆ９中进行瞬间表达，用ＩＬ－６依赖细胞株ＭＨ６０·ＢＳＦ２检测其表达活性，结果表明，感染后４８ｈ后就具有明显ＩＬ－６表达，由此可见，利用该系统可成功地表达小鼠ＩＬ－６基因。     

Abortive replication of Bombyx mori nucleopolyhedrovirus in Sf9 and High Five cells: defective nuclear transport of the virions

Despite close genetic relationship, Bombyx mori nucleopolyhedrovirus (BmNPV) and Autographa californica multicapsid NPV (AcMNPV) display a distinct host range property. Here, BmNPV replication was examined in Sf9 and High Five cells that were nonproductive for BmNPV infection but supported high titers of AcMNPV replication. Recombinant BmNPV, vBm/gfp/lac, containing bm-ie1 promoter-driven egfp showed that few Sf9 and High Five cells infected with vBm/gfp/lac expressed EGFP, while large proportion of EGFP-expressing cells was observed when transfected with vBm/gfp/lac DNA. Immunocytochemical analysis showed that BmNPV was not imported into the nucleus of these two cell lines, while recombinant BmNPV, vBmDelta64/ac-gp64 possessing AcMNPV gp64 was imported into the nucleus, yielding progeny virions in High Five cells, but not Sf9 cells. These results indicate that the defective nuclear import of infected virions due to insufficient BmNPV GP64 function is involved in the restricted BmNPV replication in Sf9 and High Five cells.     

人乳头瘤病毒16型结构基因L1在昆虫细胞中的表达及其生物学活性

目的 研究与宫颈癌及人类其它多种组织癌症密切相关的人乳头瘤病毒16型（HPV16）的晚期基因L1的表达及其生物学活性。方法 采用PCR法从pBSSK-B/16L1扩增了来自中国妇女鲍温病组织标本的HPV16晚期基因L1,装入杆状病毒转移载体;在DH10Bac内通过转座子Tn7的介导,使携带有杆状病毒多角体蛋白基因启动子Ppolh的HPV16 L1基因整合入杆状病毒,形成重组杆状病毒;转染昆虫细胞Sf9进行表达;将感染72h的Sf9细胞包埋、切片、染色后,电镜观察。提取L1蛋白,免疫BALB／ c小鼠。结果 重组杆状病毒在Sf9细胞内高效表达出L1蛋白,经Western blot发现能与L1抗体特异地结合;薄层扫描显示所表达的L1蛋白占Sf9细胞总蛋白的比例高达31％。经透射电镜观察表明,在细胞核里有大量的重组杆状病毒形成;并且产生了大量的由HPV16 L1蛋白单体自组装的病毒样颗粒（virus-like particles,VLP)。小鼠免疫实验结果表明,所表达的HPV16 L1蛋白单体自组装的病毒样颗粒（virus-like particles,VLP)。小鼠免疫实验结果表明,所表达的HPV16 L1蛋白具有强免疫原性。结论 此研究为今后L1基因分子流行病学检查、L1蛋白的结构生物学研究、疫苗研制以及HPV相关基础性研究提供了有用的资料。     

AcMNPV enhances infection by ThorNPV in Sf21 cells and SeMNPV in Hi5 cells

An expression cassette containing the DsRed2 gene, which encodes the red fluorescent protein (RFP), was inserted into the wide-host-range Autographa californica M nucleopolyhedrovirus (AcMNPV) at the polyhedrin locus (vAcDsRed2). An expression cassette containing the enhanced green fluorescent protein (EGFP) gene was inserted at the gp37 locus of the narrow-host-range Thysanoplusia orichalcea MNPV (ThorMNPV) and the p10 locus of Spodoptera exigua MNPV (SeMNPV) to produce vThGFP and vSeGFP, respectively. vThGFP and vSeGFP are poor at infecting Sf21 and Hi5 cells, respectively, whereas vAcDsRed2 is highly infectious to both cell lines. During co-infection, vAcDsRed2 enhanced vThGFP infection in Sf21 cells by approximately 20-fold, and it enhanced vSeGFP infection in Hi5 cells by more than 300-fold, as detected by fluorescence measurements. In contrast, vThGFP reduced vAcDsRed2 infection by 5.4-fold in Sf21 cells, while vSeGFP reduced vAcDsRed2 by 3.2-fold in Hi5 cells. Plaque assay data did not suggest viral recombination, but vThGFP plaques surrounded by vAcDsRed2 plaques were observed. A viral DNA replication assay performed by real-time quantitative PCR suggested that the detected fluorescence correlated with virus replication. Sf21 cells infected with vAcDsRed2 were resistant to superinfection by viruses of the same type expressing EGFP (vAcGFP). These results demonstrated that AcMNPV could enhance replication of ThorMNPV and SeMNPV in non-permissive cells without recombination.     

Modulation of dopamine D1 receptor signaling by adenosine A1 receptors in Sf9 cells requires expression of Gi proteins

An excess accumulation of advanced glycation end products (AGEs) has been reported in autism brains. Through their interaction with their putative receptor RAGE, AGEs can promote neuroinflammation, oxidative stress and neuronal degeneration. To shed more light on the possible alterations of the AGEs–RAGE axis in autism, hereto we measured plasma levels of endogenous secretory RAGE (esRAGE) and its proinflammatory ligand S100A9 in 18 young adults with autistic spectrum disorder (ASD) and 18 age- and gender-matched healthy comparison subjects. The Childhood Autism Rating Scale (CARS) was used to assess the severity of autistic symptoms. Significantly reduced levels of esRAGE (P = 0.0023) and elevated concentrations of S100A9 (P = 0.0012) were found in ASD patients as compared to controls. In autistic patients, there was a statistically significant positive correlation between CARS scores and S100A9 levels (r = 0.49, P = 0.035), but no significant correlation was seen between esRAGE and S100A9 values (r = −0.23, P = 0.34). Our results of a significantly reduced peripheral level of esRAGE coupled with elevated S100A9 point to a subtle but definite dysfunction of the AGEs/RAGE axis in autism that could play a role in the pathophysiology of this disorder.