Differential regulation of expression of the MHC class II molecules RT1.B and RT1.D on rat B lymphocytes: effects of interleukin-4, interleukin-13 and interferon-gamma.

Susceptibility to induction of both T helper 1- (Th1) and Th2-mediated autoimmunity is multifactorial and involves genetic linkage to the major histocompatibility complex (MHC) class II haplotype. Brown Norway (BN) rats exposed to mercuric chloride develop a Th2-dependent systemic autoimmunity, whereas Lewis rats, which are highly susceptible to Th1-mediated autoimmunity, develop immune suppression after mercuric chloride exposure. Exposure to mercuric chloride is known to enhance B-lymphocyte expression of the MHC class II molecule RT1.B, predominantly in BN rats. We demonstrate that, in contrast, expression of RT1.D was unmodified on these B cells, whereas both RT1.B and RT1.D were up-regulated on epithelial cells. Regulation of B-cell MHC class II isotype expression was further studied in vitro, using BN rat lymph node (LN) cells. Interleukin-4 (IL-4) strongly enhanced B-cell expression of RT1.B (2.8-fold), whereas RT1.D expression was only slightly, although significantly, modified (1.2-fold). B cells from Lewis rats showed a similar IL-4-induced enhancement of RT1.B expression (2.5-fold), whereas, in contrast, RT1.D expression was unmodified. Exposure of LN cells from BN rats to interferon-gamma induced a moderate increase of B-cell MHC class II expression, predominantly of RT1.B. Strong and rapid enhancement of B-cell RT1.D expression was observed after stimulation by phorbol 12-myristate 13-acetate and ionomycin. Rat IL-13 did not modify B-cell MHC class II expression; however, it induced typical morphological changes in peritoneal macrophages. These experiments demonstrate isotype-specific and strain-dependent regulation of MHC class II expression on rat B lymphocytes, which may be of pathophysiological relevance for the strain-dependent susceptibility for Th1- or Th2-mediated autoimmunity.     

Microarray analysis on CYPs expression in pregnant rats after treatment with pregnenolone-16alpha-carbonitrile and phenobarbital

We previously reported the protein expression profiles of nine cytochrome P450 isozymes (CYPs) in pregnant rat's liver, fetal liver, and placenta after treatment with pregnenolone-16alpha-carbonitrile (PCN), dexamethasone (DEX), or phenobarbital (PB). In this study, the gene expression of 40 CYPs and 2 orphan nuclear receptors for CYP inducers, that is, Nr1i2 (CYP3A subfamily inducible by PCN) and Nr1i3 (CYP2B subfamily inducible by PB), in pregnant rat's liver, fetal liver, and placenta was investigated at one time. Fischer 344 (F344) pregnant rats were daily treated intraperitoneally with 50 mg/kg of PCN or 80 mg/kg of PB from 13 to 16 days of gestation (DG). They were sacrificed on 17 DG, and microarray analysis using Affymetrix Rat Expression Array 230A was performed. Ten genes expression significantly increased in dam's liver in PCN group, and seven genes expression in PB group. On the other hand, four genes expression increased in fetal liver in PCN group, and three genes expression increased in PB group. Being common to dam's and fetal livers, the gene expression of Cyp3A1 (CYP3A subfamily) and cytochrome P-450e (CYP2B subfamily) increased in both PCN and PB groups. In placenta, the expression of Cyp3A1 gene was significantly induced in PB group, and it also showed a tendency to increase in PCN group. The expression of Nr1i2 gene was significantly elevated only in dam's liver of PCN group, while the expression of Nr1i3 gene showed no changes in all groups. The results of the present study of 40 CYPs gene expression mostly corresponded to our previous reports on 9 CYPs protein expression.     

CD86协同刺激信号在孕早期母—胎界面Th1／Th2型细胞因子表达调控中的作用

目的探讨CD86协同刺激信号在孕早期母-胎界面Th1/Th2型细胞因子表达调控中的作用。方法建立正常妊 娠模型CBA×Balb/c和自然流产模型CBA× DBA/2。于孕第4、6、8、10天给CBA孕鼠腹腔注射大鼠IgG作为对照组;仅于孕第4 天或于孕第4、6、8、10天给CBA孕鼠腹腔注射大鼠抗小鼠CD86 mAb。孕第14天计算两种模型各实验组胚胎吸收率R。ELISA 法测定孕第9和第14天各实验组母-胎界面组织体外培养上清中Th1/Th2型及相关细胞因子(IL-2、IFN-γ、TNF-α、IL-4、IL-10、 TGF-β2)表达水平。结果孕早期干预CD86协同刺激信号,对正常妊娠模型母-胎界面孕第9和第14天IL-4、IL-10、TGF-β2以及 IFN-γ、TNF-α表达均无显著影响,其胚胎吸收率亦无显著变化。自然流产模型中,孕早期干预CD86协同刺激信号后,孕第9、 第14天母-胎界面IL-4、IL-10、TGF-β2表达均显著增加(P＜0.05);而IFN-γ、TNF-α表达显著下降(P＜0.05)。胚胎吸收率亦 显著下降(P＜0.05)。结论孕早期母-胎界面CD86协同刺激信号的调节紊乱可能是触发母体对胚胎产生免疫排斥的重要病 理因素,于孕早期干预CD86协同刺激信号能够恢复母-胎界面Th1型/Th2型免疫调节的生理平衡,从而诱导母-胎免疫耐受。     

早孕期弓形虫感染对大鼠胎盘RT.BMl及RTl-E基因转录的影响

通过检测正常妊娠和早孕期弓形虫感染的Wister大鼠胎盘RT.BMl和RTl-E mRNA的表达水平,发现二者在正常妊娠中的规律,进而探索早孕期弓形虫感染对胎盘RT.BMl和RTl-E mRNA表达的影响,以阐明弓形虫感染致不良妊娠结局的分子免疫机制.将48只孕鼠随机分为感染组和正常组,每组24只.感染组于妊娠第5天腹腔注射1×105个弓形虫强毒株(RH株)速殖子/只,正常组不作任何处理.两组于孕第9、13、15、17天分别处死6只孕鼠,无菌剖腹,观察胎鼠情况,计数死胎及活胎数.采用实时荧光定量聚合酶链式反应(real-time PCR)检测胎盘RT.BMl和RTl-E mRNA表达水平.结果:①正常组活胎数204只,死胎3只,死胎率1.4%,感染组活胎数133只,死胎27只,死胎率16.9%,死胎率明显高于正常组(P<0.05);②正常组RT.BMl mRNA在孕第9、13、15、17天的相对表达水平为(3.21±1.32)、(4.35±1.29)、(3.96±1.49)、(11.1±2.15),前三个时间点表达水平基本一致(P>0.05),第17天则明显增高(P<0.01);RTl-E mRNA在各时间点相对表达水平为(0.79±0.45)、(2.15±1.03)、(2.13±1.74)、(2.394±1.85),第9天明显低于后三个时间点(P<0.05);③感染组各时间点RT.BMl mRNA相对表达水平为(12.4±2.86)、(11.5±3.67)、(6.89±1.59)、(17.3±3.06),第9、13、15天与正常组相比有显著差异(P<0.01);而RTl-E mRNA分别为(1.834±0.886)、(4.72±1.59)、(4.24±2.26)、(3.83±2.26),第9、13、15天与正常组相比均有显著差异(P<0.01)、(P<0.05)、(P<0.01).结果显示弓形虫感染量合适,动物模型成功建立;RT.BMl和RTl-E mRNA在大鼠胎盘的表达水平与妊娠进展密切相关;大鼠早孕期弓形虫感染可致胎盘局部RT.BMl和RTl-E表达增高,从而打破正常妊娠所需的胎蕊局部免疫平衡状态,可能是早孕期弓形虫感染致不良妊娠结局的分子机制之一.     

早孕期弓形虫感染对大鼠胎盘RT.BM1及RT1-E基因转录的影响

通过检测正常妊娠和早孕期弓形虫感染的Wister大鼠胎盘RT.BM1和RT1-E mRNA的表达水平,发现二者在正常妊娠中的规律,进而探索早孕期弓形虫感染对胎盘RT.BM1和RT1-E mRNA表达的影响,以阐明弓形虫感染致不良妊娠结局的分子免疫机制。将48只孕鼠随机分为感染组和正常组,每组24只。感染组于妊娠第5天腹腔注射1×105个弓形虫强毒株(RH株)速殖子/只,正常组不作任何处理。两组于孕第9、13、15、17天分别处死6只孕鼠,无菌剖腹,观察胎鼠情况,计数死胎及活胎数。采用实时荧光定量聚合酶链式反应(real-time PCR)检测胎盘RT.BM1和RT1-E mRNA表达水平。结果:①正常组活胎数204只,死胎3只,死胎率1.4%,感染组活胎数133只,死胎27只,死胎率16.9%,死胎率明显高于正常组(P<0.05);②正常组RT.BM1 mRNA在孕第9、13、15、17天的相对表达水平为(3.21±1.32)、(4.35±1.29)、(3.96±1.49)、(11.1±2.15),前三个时间点表达水平基本一致(P>0.05),第17天则明显增高(P<0.01);RT1-E mRNA在各时间点相对表达水平为(0.79±0.45)、(2.15±1.03)、(2.13±1.74)、(2.39±1.85),第9天明显低于后三个时间点(P<0.05);③感染组各时间点RT.BM1 mRNA相对表达水平为(12.4±2.86)、(11.5±3.67)、(6.89±1.59)、(17.3±3.06),第9、13、15天与正常组相比有显著差异(P<0.01);而RT1-E mRNA分别为(1.83±0.886)、(4.72±1.59)、(4.24±2.26)、(3.83±2.26),第9、13、15天与正常组相比均有显著差异(P<0.01)、(P<0.05)、(P<0.01)。结果显示弓形虫感染量合适,动物模型成功建立;RT.BM1和RT1-E mRNA在大鼠胎盘的表达水平与妊娠进展密切相关;大鼠早孕期弓形虫感染可致胎盘局部RT.BM1和RT1-E表达增高,从而打破正常妊娠所需的胎盘局部免疫平衡状态,可能是早孕期弓形虫感染致不良妊娠结局的分子机制之一。     

Effect of a serotonin 1-A agonist on food intake of Osborne-Mendel and S5B/P1 rats

The effect on food intake of 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT), a serotonin 1-A agonist, has been evaluated in two strains of rats that differ in their sensitivity to becoming obese while eating a high-fat diet. Male Osborne-Mendel (OM) and S5B/Pl rats were tested at 8 weeks and 16 weeks of age. Both strains were adapted to choose between two diets-a 56% fat energy diet, and a 10% fat energy diet-which were equicaloric for protein (24% energy). Daily food intake was measured for 2 weeks before injection of 8-OH-DPAT. The younger OM rats had no diet preference, while the older OM rats preferred the high-fat diet. The younger S5B/P1 rats preferred the low-fat diet, while the older S5B/P1 rats had no diet preference. Satiated rats were injected subcutaneously with 8-OH-DPAT at doses of 0.3, 1.0, or 3.0 mg/kg or vehicle. During the light phase, subcutaneous 8-OH-DPAT increased the intake of the high-fat diet in the 16-week-old OM rats but not the 8-week-old OM rats. 8-OH-DPAT had no effect on the low-fat diet intake in either age OM rat. The 8-week-old S5B/P1 rats showed a significant decrease in intake of the high-fat diet in response to 8-OH-DPAT, whereas an increase in the intake of the low-fat diet was observed in the older S5B/P1 rats. These data are consistent with the hypothesis that increased fat preference in Osborne-Mendel rats may result in part from altered serotonin activity of 5-HT(1A) receptors.     

The Fc receptor, FcRI, and other activation molecules on human mononuclear phagocytes after treatment with interferon-gamma.

Human monocytes and the myeloid cell lines U937 and HL60 have been tested with monoclonal antibodies (MoAbs) reactive with 22 different cell surface molecules before and after treatment with interferon-gamma (IFN-gamma). An increase in the expression of the high affinity Fc receptor, FcRI, and the receptor for interleukin 2, IL-2R, were the most consistent alterations which were observed. In addition, expression of the gp55 molecule recognized by CD14 MoAbs was decreased on monocytes. Of the MHC Class II molecules, there was little expression by the myeloid cell lines and no enhancement after IFN-gamma treatment. In contrast monocytes expressed all three MHC Class II subloci with DR much greater than DQ and DP. However there was much variation in IFN-gamma-mediated increase in expression of the individual subregions. In monocytes, the alteration in expression of FcRI, IL-2R, gp55 and MHC Class II molecules took place in a co-ordinate fashion and reached a plateau only after 48 h. In U937 cells, activation proceeded more rapidly and was at maximum levels between 12-16 h. This increase in FcRI appears to be a hallmark of IFN-gamma activation for mononuclear phagocytes (Mph) as the other alterations are either not found on all types of Mph (gp55, MHC Class II) or are induced by other cytokines on Mph and on other cells (IL-2R, MHC Class II). Conversely, other cytokines do not induce FcRI on Mph. These results also suggest that the cell membrane phenotypic changes induced in Mph by IFN-gamma may not be extensive and that FcRI must play a specific role in the IFN-gamma-activated Mph.     

Induction of HLA-DR expression on human myoblasts with interferon-gamma

Myoblast transplantation is an experimental therapy that may be useful in hereditary muscle diseases. One obstacle to this approach is immune rejection. We evaluated human myoblasts and myotubes for surface expression of HLA class I and II histocompatibility antigens. The myoblast preparations, cultured from muscle biopsy specimens, were more than 95% pure as assessed by immunostaining with the Leu19 monoclonal antibody that reacts with human myoblasts and regenerating muscle fibers. Myoblasts and myotubes constitutively expressed HLA-class I but not HLA-class II molecules. However, HLA-class II expression was induced on mononucleated myoblasts after culture for 5 days in the presence of recombinant human interferon-gamma. The results indicate that transplanted myoblasts can be rejected because of histoincompatibility at the HLA-class I and HLA class II locus. Furthermore, an aberrant expression of HLA-class II antigen on myoblasts in vivo may play a role in the pathogenesis of autoimmune muscle disorders.     

Curcumin 对注射脂多糖大鼠肺脏血红素氧合酶-1表达的影响

目的：探讨活化蛋白-1（AP-1）阻断剂curcumin对注射脂多糖大鼠肺组织血红素氧合酶（HO-1）表达的转录调节机制。 方法： 18只大鼠随机分为3组：对照组（经舌静脉注入等量生理盐水）、脂多糖（LPS）组（经舌静脉注入LPS 5 mg·0.5 mL-1·kg-1）和LPS+curcumin组（经腹腔注入AP-1阻断剂curcumin 20 mg·0.5mL-1·kg-1，20 min后再注入LPS 5 mg·0.5 mL-1·kg-1）。给药7 h后杀死动物留取肺组织，应用逆转录聚合酶链反应（RT-PCR）和Westren 印迹法分别检测HO-1 mRNA和蛋白的表达，生化方法检测肺组织匀浆碳氧血红蛋白（HbCO）含量，间接代表肺组织中一氧化碳（CO）含量。 结果： LPS组大鼠肺组织HO-1 mRNA和蛋白的表达水平及肺组织CO含量高于对照组（均P<0.01），而LPS+curcumin组肺组织HO-1 mRNA、蛋白表达及CO含量明显低于LPS组（均P<0.01）。 结论： LPS攻击的大鼠肺组织中HO-1基因转录可能是通过激活AP-1进行调控的。     

肠源性内毒素血症对肝组织细胞间粘附分子-1表达的影响

目的:探讨肠源性内毒素血症对肝组织细胞间粘附分子-1(ICAM-1)表达的影响。方法:采用Western blot 的方法检测硫代乙酰胺(TAA)诱导的肠源性内毒素血症大鼠肝组织中ICAM-1表达的变化。结果:ICAM-1的分子量为95 kD,正常对照组与注射TAA 6 h组ICAM-1的表达很弱,而在注射TAA 12 h以后的表达有增强,且与血浆内毒素水平及反映肝损伤程度的ALT活性变化相一致。结论:肠源性内毒素血症可诱导肝组织中ICAM-1表达的上调,后者与肝损伤程度相一致。     

小檗碱对淋巴细胞与血管内皮细胞粘附及粘附分子的影响

目的：通过研究小檗碱对淋巴细胞与血管内皮细胞粘附及粘附分子的影响,探讨小檗碱免疫抑制作用机制。方法：以小檗碱处理人淋巴细胞或人脐静脉内皮细胞（ＨＵＶＥＣ）后,用蛋白染料染色法研究对淋巴细胞与内皮细胞粘附的作用,用细胞ＥＬＩＳＡ,免疫细胞化学染色法研究对细胞表面粘附分子表达的影响。结果：小檗碱能抑制静息的及ＩＬ－１,ＴＮＦ激活的内皮细胞与淋巴细胞间的粘附,其主要分子机制为下调内皮细胞表面粘附分子ＩＣ     

Microarray analysis on Phase II drug metabolizing enzymes expression in pregnant rats after treatment with pregnenolone-16alpha-carbonitrile or phenobarbital

We previously reported the expression profiles of 9 cytochrome P450 isozymes (CYPs) proteins and those of 40 CYPs genes in pregnant rat's liver, placenta and fetal liver after treatment with pregnenolone-16alpha-carbonitrile (PCN) or phenobarbital (PB). This study was carried out focusing on the gene expression profiles of Phase II drug metabolizing enzymes, Glutathione S-transferase isozymes (GSTs) and UDP-glycosyltransferase isozymes (UDPGTs). Fischer 344 (F344) pregnant rats were daily treated intraperitoneally with 50 mg/kg of PCN or 80 mg/kg of PB from 13 to 16 days of gestation (DG). They were sacrificed on 17 DG, and microarray analysis using Affymetrix Rat Expression Array 230 A was performed. Among 16 GSTs genes examined in this study, 7 genes were significantly induced in dam's liver and 3 genes in fetal liver, respectively, in the PCN-group, while 8 genes were significantly induced in dam's liver and 1 gene in fetal liver, respectively, in the PB-group. On the other hand, among 11 UDPGTs genes examined, 5 genes were significantly induced in dam's liver and 3 genes in fetal liver, respectively, in the PCN-group, while 5 genes were significantly induced in dam's liver and 1 gene in fetal liver, respectively, in the PB-group. There were no significant changes in the placenta of all groups. This is the first report of the gene expression profiles of Phase II drug metabolizing enzymes in pregnant rat and fetal livers and placenta after treatment with typical inducers of drug metabolizing enzymes.     

阻断协同刺激分子对TGF-β1和PAI-1的表达及妊娠结局的影响

目的：探讨阻断CD86协同刺激分子对自然流产模型孕鼠TGF-β1和PAI-1的表达及妊娠结局的影响。方法：实验组于妊娠第4.5天腹腔注射大鼠抗小鼠CD86单抗，实验对照组注射大鼠同型IgG2b，而正常妊娠组不做任何处理。于妊娠第13.5天计算胚胎吸收率，用免疫组化测定TGF-β1和PAI-1的表达。结果：（1）实验组的胚胎吸收率显著低于实验对照组（P〈0.05）。（2）实验组蜕膜细胞中TGF-β1和PAI-1的表达均显著高于实验对照组（P〈0.05）。结论：妊娠早期阻断CD86协同刺激分子信号途径可使母胎界面中的TGF-β1和PAI-1分别通过各自不同的途径来发挥免疫耐受作用并且使自然流产模型孕鼠的胚胎吸收率降低至正常妊娠水平。     

Effect of phenformin on lipid and carbohydrate metabolism in pregnant rats

A decrease in the blood levels of cholestrol, phospholipids, and free fatty acids was observed in rats receiving the biguanide phenformin (5–25 mg daily by mouth) from the first to eighth day of pregnancy, but no developmental anomalies of the fetuses or placenta were found. The acceptability of biguanide administration during pregnancy is discussed.Presented by Academician of the Academy of Medical Sciences of the USSR A. I. Serebrov.Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 82, No. 7, pp. 825–827, July, 1976.