Effect of Topically Applied Cyclosporin A on Arachidonic Acid (AA)- and Tetradecanoylphorbol Acetate (TPA)-Induced Dermal Inflammation in Mouse Ear

Topical cyclosporin A (CsA) was compared with dexamethasone, indomethacin and phenidone in edema, increases in vascular permeability, eicosanoids and cell-influx induced by arachidonic acid (AA) and tetradecanoylphorbol acetate (TPA) in mouse ears. CsA ED₅₀ on AA-edema (7.7 g/ear) was similar to dexamethasone and lower than indomethacin and phenidone. CsA ED₅₀ in TPA edema (21 g/ear) was higher than dexamethasone and lower than indomethacin or phenidone. All drugs equally reduce the AA-induced increase in vascular permeability, but CsA and dexamethasone had more activity on TPA. AA-increase in 6-keto-PGF₁ was reduced by dexamethasone, indomethacin and phenidone but not by CsA; only phenidone reduced LTB₄. TPA-increase in 6-keto-PGF₁ was reduced by CsA and indomethacin while CsA, dexamethasone and phenidone decreased LTB₄. CsA, indomethacin and phenidone, but not dexamethasone, suppressed AA-neutrophil influx. In TPA-ears all drugs produced similar reduction in neutrophil influx. CsA was shown to be a good topical anti-inflammatory drug.     

Effect of Topically Applied Cyclooxygenase-2-Selective Inhibitors on Arachidonic Acid- and Tetradecanoylphorbol Acetate-Induced Dermal Inflammation in the Mouse

The topical effects of cyclooxygenase-2 (COX-2)-selective inhibitors, flosulide (CGP 28238), L-745,337 and SC-57,666 were examined in AA- and TPA-induced ear dermal inflammation in the mouse. The doses that caused 50% inhibition in AA edema (ED₅₀) were 2.4, 0.45 and 0.35 mg/ear for flosulide, L-745,337 and SC-57,666, respectively. The respective ED_50s in TPA-edema were 1, 0.45 and 0.14. Indomethacin and zileuton showed higher activity than the COX-2-selective inhibitors in both models. Flosulide and L-745,337 inhibited the AA-induced increase in 6-keto-PGF₁, while SC-57,666 was inactive. 80% inhibition was seen with indomethacin while zileuton had no effect. COX-2 selective inhibitors and indomethacin had no effect on LTB₄ levels, while zileuton produced a 50% inhibition. The TPA-induced increase in 6-keto-PGF₁ was greatly inhibited by all COX-2 inhibitors while LTB₄was potentiated by both flosulide and L-745,337. Indomethacin inhibited 6-keto-PGF₁ and zileuton reduced 6-keto-PGF and strongly reduced LTB₄. The neutrophil influx induced by AA was lower than that of TPA. Myeloperoxidase (MPO) levels were lowered by flosulide and L-745,337 but not by SC-57,666. TPA-induced MPO increase was decreased by all COX-2 inhibitors. Indomethacin and zileuton had similar effect on AA and TPA-induced increase in MPO. The results indicate that COX-2-selective inhibitors showed lower topical anti-inflammatory activity than indomethacin or zileuton.     

经颞骨岩部乙状窦前入路处理岩斜区病变的应用解剖

目的 :研究经颞骨岩部乙状窦前入路进行岩斜区直接手术的方法 ,寻找出该入路的标志点和颞骨岩部的磨除方法 ,并观察手术的暴露范围和优缺点。方法 :用 15例血管经彩色乳胶灌注的成人尸头标本 ,显微镜下 (× 6～ 2 5 )解剖经颞骨岩部乙状窦前入路。结果 :岩乙状窦交叉点、内淋巴囊裂是磨除颞骨岩部后面的重要标志 ,用内淋巴囊裂可以初步定位前庭小管、总脚和后半规管。此入路能够暴露从鞍背到延髓中上部的区域 ,以暴露中岩斜区最佳。离断内淋巴囊 ,可扩大骨磨除的面积 ,增加手术的暴露。结论 :以岩乙状窦交叉点、内淋巴囊裂为标志为安全磨除颞骨岩部后面提供了保证。经颞骨岩部乙状窦前入路具有手术距离短、视野暴露充分的特点 ,特别适用于骑跨颅中、后窝而以颅后窝为主的岩斜区病变     

Comparative evaluation of arachidonic acid (AA)- and tetradecanoylphorbol acetate (TPA)-induced dermal inflammation

The effects of topical application of arachidonic acid (AA) or phorbol ester, tetradecanoylphorbol 13-acetate (TPA), on edema response, vascular permeability, MPO, NAG, and generation of eicosanoids were studied in two murine models of cutaneous inflammation. AA produced a short-lived edema response with a rapid onset that was associated with marked increases in levels of prostaglandins (PGE₂, 6-keto-PGF₁, PGF₂), thromboxane B₂ (TxB₂) and leukotriene B₄ (LTB₄), with smaller increases in levels of LTC₄. TPA produced a longer-lasting edema that was associated with marked influx of neutrophils and predominant formation of LTB₄ along with significant changes in levels of TxB₂. Circulating T lymphocytes have no apparent role in the acute inflammatory responses induced by either agent. Arachidonic acid-induced vascular permeability preceded the edema response and neutrophil influx, whereas TPA-induced vascular permeability paralleled the edema response and influx of neutrophils. Mast cells appear to be important in the complete expression of inflammatory response, i.e., edema, cellular influx, and vascular permeability induced by either AA or TPA, as these responses were blunted in mast cell-deficient mice. Inhibitors of CO or 5-LO attenuated inflammatory responses in both models. The LTB₄ receptor antagonist, SC-41930, inhibited the inflammatory response to TPA but had little effect on that initiated by AA. This suggests that LTB₄ is an important mediator in the phorbol ester-induced inflammatory response, whereas peptidoleukotrienes and prostaglandins regulate vascular permeability responses in the arachidonate model.     

失血性休克对脾脏细胞因子和花生四烯酸代谢的影响

目的：探讨脾脏在失血性休克时对脾脏细胞因子和花生四烯酸代谢的调节作用,方法：分别失血性休克家犬和家兔动脉血、脾静脉血和脾组织均浆放射免疫法测定其IL－1、IL－2、IL－6、IL－8、TNF、TXB2和6KPGF1α含量.结果：1．失血性休克后动脉血浆中IL－1、IL－6、IL－8、TNF、含量显著降低,而IL－2、TXB2和6KPGF1α含量明显增高,2．失血性休克后脾静脉血浆中IL－1、IL－2,IL－6,IL－8、TNF、6KPGF1α含量明显降低,仅TXB2含量明显增高,而休克治疗后IL－8TNF、6KPGF1α含量均明显产高;3．失血性休克后脾组织中IL－1和TXB2含量降低,IL－2、IL－6、TNF、6KPGF1α含量明显增高;4．切脾对TXB2生成无影响,而对6KPGF1α的生成有一定影响,结论：脾及对失血性休克时的多种功能因子的生成和释放有一定作用,对花生四烯酸代谢也有一定影响。     

The Presynaptic Effects of Arachidonic Acid and Prostaglandin E2 at the Frog Neuromuscular Junction

Arachidonic acid and prostaglandin E₂ decreased the frequency of miniature endplate potentials with producing any changes in the their amplitude-time parameters. Arachidonic acid and prostaglandin E₂ decreased the quantum composition of endplate currents and the amplitude of the third phase of the nerve ending response, which reflects currents though potential-dependent K⁺ channels. A perineural method was used to demonstrate that arachidonic acid and prostaglandin E₂ suppressed the nerve ending Ca²⁺ current. The cyclooxygenase blocker indomethacin increased neurotransmitter secretion and decreased the third phase of the nerve ending response. The effects of arachidonic acid and prostaglandin E₂ on evoked neurotransmitter release were not seen in the presence of indomethacin, while the third phase of the response continued to show a reduction. It is suggested that prostaglandin E₂ mediates the effects of arachidonic acid on spontaneous and evoked neurotransmitter secretion, Ca²⁺ currents, and Ca²⁺-dependent K⁺ currents. In addition, arachidonic acid and prostaglandin E₂ had their own effects on potential-dependent K⁺ currents in nerve endings. __________ Translated from Rossiiskii Fiziologicheskii Zhurnal imeni I. M. Sechenova, Vol. 91, No. 3, pp. 268–276, March, 2005.     

Effects of Castration, Depo-Testosterone and Cyproterone Acetate on Lymphocyte T Subsets in Mouse Thymus and Spleen

The effects of testosterone on the relative proportion of Thy 1.2, CD4 (L3T4) and CD8 (Lyt-2) cells in thymus and spleen were studied after castration and administration of Depo-testosterone (DT) separately or together with cyproterone acetate (CA) (an antiandrogen) in BDF1 mice. Injection of 0.5 mg/100 g body weight of DT during 2 weeks decreased significantly the number and proportion of double positive (DP) (CD4+ CD8+) and increased the percentage of single positive (SP) CD4+ (CD4+ CD8-), whereas there was a slight decrease in the Thy 1.2+ cells in the thymus. In parallel, we observed an increase in CD8+ (CD4- CD8+) cells in the spleen. The androgen deprivation after 3 weeks of castration induced a decrease in the percentage of CD4+ cells in thymus and both CD4+ and CD8+ cells in spleen. Injection of CA (0.5 mg/100 g body weight) had the same qualitative effects as DT on the proportion of lymphocyte T subsets in castrated mice. However, the combined activities of DT and CA were greater than either alone. These data indicate the main role of testosterone in the distribution of CD4+ and CD8+ cells in male mice. The similar effects of CA and DT in the lymphoid organs may suggest a difference between androgen receptors of sexual and lymphoid organs.     

Effects of a protein kinase C inhibitor (PKCI) on the development of adjuvant-induced arthritis (AA) in rats

Inflammation Research -     

Effects of 12-o-tetradecanoyl-phorbol-13-acetate (TPA) on the colony growth of human T lymphocytes

The T colony promoting activity of 12-o-tetradecanoyl-phorbol-13-acetate (TPA) was assessed in a double layer culture assay which is dependent on the simultaneous presence of phytohaemagglutinin (PHA) and a leucocyte rich underlayer. TPA (10(-8) M) incorporated in the overlayer in place of PHA was capable of promoting T cell growth in the form of clusters in all 37 experiments performed and in the form of colonies in more than 50% of the samples tested. However, the T colony promoting activity of TPA alone was markedly less evident and consistent than that of PHA (mean 13 +/- 19.9 s.d. colonies vs 168 +/- 78.6). TPA concentrations of 10(-6) M, 10(-9) M and 10(-10) M were practically ineffective. On the other hand, the number of colonies obtained when both TPA 10(8) M and PHA were incorporated in the overlayer was significantly higher (P less than 0.01) than that observed with PHA alone (mean 250 +/- 108.2 vs 178 +/- 84.5 colonies). When TPA concentrations of 10(-9) M and 10(-10) M were used in addition to PHA, the enhancing effect was less evident, while an inhibition of T colony growth was observed with TPA 10(-6) M + PHA. TPA 10(-8) M was also capable of enhancing T colony growth when incorporated in the leucocyte rich underlayer (222 +/- 98.6 vs 172 +/- 80.9 colonies). In all cultures with TPA the peak of growth was delayed compared with that of control experiments with PHA. These findings demonstrate that TPA, particularly when co-cultured with PHA, is an effective T colony promoting agent. The observation that the number of colonies formed in the presence of TPA plus PHA is higher than the sum of those observed with the two stimulators independently, suggests that their synergistic effect may be mediated via the production of colony stimulating soluble factors.     

卡芬尼对大鼠佐剂性关节炎治疗作用的免疫机制

本文研究了卡芬尼(CCA)治疗大鼠佐剂性关节炎(AA)的免疫调节机制。CCA 能使 AA大鼠低下的脾细胞刀豆素 A(ConA)反应、IL—2产生能力恢复正常,使外周血中减少的 Ts 细胞数目回升到正常水平,使 AA 大鼠腹腔 M(?)升高的 IL—1和 H_2O_2产生水平降至正常范围,还能明显减少 AA 大鼠炎性关节组织中 PGE_(?)的含量.研究 CCA 对体外正常大鼠淋巴细胞与 M(?)以及 AA 大鼠 M(?)功能的影响时发现,CCA 对 ConA 诱导的脾细胞增殖反应呈低浓度(0.1～1μg/ml)增强和高浓度(10μg/ml)抑制的双向作用;CCA(100μg/ml)对常大鼠腹腔 M(?)释放 H_2O_2无明显影响,但能显著减少 AA 大鼠腹腔 M(?)释放 H_2O_2.结果表明,CCA 对 AA 大鼠有明显的免疫调节作用,这种作用有明显的机能和剂量依赖性.CCA 对 AA 大鼠的治疗作用与其调整患鼠异常的免疫功能有关。     

Effects of n-Butyrate and phorbol ester (TPA) on induction of Epstein-Barr virus antigens and cell differentiation

Summary N-Butyrate, an effective inducer of synthesis of Epstein-Barr virus (EBV) antigens in virus-producer P3HR-1 cells, has recently been shown (2) to induce morphological differentiation towards plasma cell in nonproducer Raji cells. The effects of n-butyrate and 12-O-tetradecanoyl-phorbol-13-acetate (TPA) on both EBV-antigen induction and cell differentiation in two virus-nonproducer lymphoblastoid cell lines, Raji and NC37, were now studied. The following observations were made (1). On its own either drug induced 1–2 per cent of cells to EBV-early-antigen positivity in both lines; their mixture induced 35 and 15 per cent positive cells in Raji and NC37 respectively (2). In Raji, n-butyrate induced about 80 per cent of cells to differentiate to plasmablast or plasma cell morphology, whereas TPA only induced the early stages of differentiation in 8 per cent of cells; a mixture of both inducers produced a similar effect as TPA alone. The addition of TPA alone or butyrate-TPA mixture led to some cellular alterations resembling virus-specific changes in virus-producer cell lines. In NC37, either drug alone or their mixture drove 13 per cent of cells to differentiate into plasmablasts or earlier stages of differentiation. In the presence of TPA protrusions and loops were seen on cell surfaces.Evidently, the stage of differentiation at which B-lymphoblastoid cell lines have been arrested can be changedin vitro. However, cell-line dependent and inducer-dependent differences in the differentiation response were apparent.With 5 Figures     

油酸多相脂质体（139）对小鼠腹腔巨噬细胞产生TNF及脾脏NK细胞活性的影响

本工作证明，油酸多相脂质体（１３９）能显著促进ＭΦ产生ＴＮＦ，同时对脾脏ＮＫ活性也有提高作用。此外，在荷瘤鼠ＮＫ活性低于正常鼠的情况下，腹腔注射１３９可在一定程度上提高荷瘤鼠的ＮＫ活性。据此推论，活化ＭΦ是１３９主要的抗肿瘤途径。     

中药风湿宁对佐剂性关节炎大鼠关节滑膜组织ICAM-1表达的影响

目的：探讨中药复方风湿宁（FSN）N-实验性类风湿性关节炎（RA）的疗效及作用机理。方法：以佐剂性关节炎（adjunvant arthritis,AA）大鼠为实验对象,分别予以FSN与雷公藤多甙（TWP）进行治疗,对照观察大鼠关节肿胀程度,以及关节内病理组织学变化情况。采用免疫组织化学检测各组AA大鼠病变关节滑膜组织中ICAM-1的表达。结果：FSN可显著抑制AA大鼠足跖肿胀,明显减轻病变关节滑膜炎症与增生,防止关节软骨及骨质的破坏;FSN能明显降低ICAM-1在AA大鼠滑膜组织的表达（P〈0.01）结论：FSN具有改善实验性RA症状及病情的作用,其抗炎作用与调控滑膜细胞-细胞间及细胞-细胞外基质间的粘附作用有关。     

HLA and Autoimmunity in Scleroderma (Systemic Sclerosis)

Scleroderma or systemic sclerosis (SSc) has been associated with certain class II antigens of the major histocompatibility complex (MHC), including HLA-DR1, DR2, DR3, DR5, and DR52. In general, these earlier HLA correlations were weak and varied considerably among reporting centers and different ethnic populations. More recently, a variety of disease-specific autoantibodies have been discovered including anticentromere, anti-topoisomerase I, and a variety of anti-nucleolar antibodies. These specificities show little overlap among one another, and each are markers for certain clinical features of SSc.

At the same time, molecular studies of the MHC have provided more accurate methods for defining specific HLA alleles. Now it is becoming clear that certain HLA class II alleles, especially HLA-DQ, are more strongly associated with autoantibody subsets of SSc than with the disease itself. For example, anticentromere antibodies are strongly associated with HLA-DQB1*0501 (DQ5), DQB1*0301 (DQ7) and other DQB1 alleles possessing a glycine or tyrosine residue in position 26 of the outermost domain. Anti-topoisomerase I antibodies occur in SSc patients with HLA-DQB1*0301 (DQ7), DQB1*0302 (DQ8), DQB1*0601 (DQ6 in Japanese), and other DQB1 alleles possessing a tyrosine residue in position 30. HLA-DQ alleles associated with these autoantibodies tend to be in linkage disequilibrium with the HLA-DR specificities previously associated weakly with SSc itself. Rare multiplex families with SSc also show these same HLA haplotypes co-segregating with autoantibody profiles in affected members. Thus, it appears that MHC alleles play a role in affecting the serological expression of SSc, and the implications of these recent findings are discussed.     

The Effect of Prior Dosing with Phenobarbitone and β-Diethylaminoethyl Diphenylpropyl Acetate (SKF525A) on the Toxicity and Liver Lesion Caused by Ngaione in the Mouse

Evidence is presented that the sesquiterpene essential oil ngaione needs metabolism in the liver for hepatotoxicity in mice. Prior treatment with SKF525A caused an increase in the LD₅₀ from 0·28 to 0·53 g/kg body weight. Paradoxically, however, pretreatment with phenobarbitone also caused an increase in the LD₅₀ to 0·37 g/kg body weight. The location of the zonal liver lesion due to ngaione, namely confluent midzonal in controls, could be consistently varied with induced alterations in the level of the hepatic mixed function oxidases in that with prior dosing with SKF525A the lesion was centrilobular, while in phenobarbitone treated mice it was periportal. These effects returned to the control state by 4 days after the respective pretreatment by which time hepatic microsomal enzyme activity was also normal.The observations suggest a pathogenetic mechanism for the rare lesion, confluent midzonal necrosis due to ngaione in this species based on graded efficiency of metabolism of the agent across the hepatocytes of the liver acinus.     

小鼠主要组织相容性复合体在移植瘤核酸水平表达情况的研究

目的：研究小鼠主要组织相容性复合体在移植肿瘤核酸及蛋白水平的表达。方法：各组织标本均采用Trizol法提取mRNA,引入细胞管家基因β-actin作为内参照物,通过RT法获取cDNA,多重PCR法同时扩增387bp的H-2抗原和550bp的β-actin。产物经1．2％琼脂糖凝胶电泳并照相存盘,半定量标本H-2和β-actin。的量,并进行各组问比较。结果：小鼠肝脏移植肿瘤H-2分子在核酸水平与正常肝脏相比,未见明显量的差异。结论：小鼠肝脏移植性肿瘤H-2的表达在核酸水平的表达量并未出现下调。