阿糖胞苷引起毛囊损伤离体模型的建立以及他克莫司对其的逆转作用

目的　建立阿糖胞苷(Ara-c)诱导毛囊损伤的离体器官培养模型,并观察他克莫司(FK506)对Ara -c诱导的小鼠触须毛囊损伤的逆转作用。方法　用离体器官培养的方法在倒置显微镜底下每日测量空白组、FK506+Ara -c和单纯Ara c作用下毛囊生长长度、记录生长天数,以及液相闪烁仪测量同位素³H-TdR的掺入率。结果　Ara-c10mg/L和25mg/L的浓度,作用2. 5h能明显抑制毛囊生长和DNA合成,缩短毛囊在体外的生长时间,抑制毛囊球部细胞增殖。0.01 ～0. 3mg/L的FK506能改善10mg/Lara -c引起的毛囊生长和DNA合成的抑制,体外生长时间缩短以及毛囊球部细胞增殖抑制。0. 1mg/L的FK506对25mg/Lara -c引起的毛囊损伤也有相似的改善作用。结论　Ara- c诱导毛囊损伤的离体模型可以用于研究化疗脱发的机制和某些因子及药物对它的干预作用,FK506在体外对Ara- c诱导的毛囊损伤有修复作用,是治疗化疗后脱发的潜在药物。     

FK506和雷怕霉素:免疫应答的新药理探针

免疫系统是尚未完全了解的复杂的网络系统,需要新工具以揭示这一网络的最小组分间的关系。FK506和雷怕霉素在临床上可能用作选择性免疫抑制剂,因而受到重视。最近两年发现两种具有免疫抑制活性的大环内酯物——FK506和雷怕霉素在延长动物同种移植器官的存活和阻止自身免疫病发作方面极为有效。两种药物都抑制T细胞增殖,但作用于T细胞活化的不同阶段,与环孢菌素A类似,FK506在早期抵消促有丝分裂或抗原刺激,抑制细胞从静止的G_0期进入细胞周期的G_1期,而雷怕霉素干扰与DNA合成关系更为密切的过程,在G_1后期、S期前抑制细胞分裂。两者很可能具有不依赖于DNA复制激活     

中药有效成分Nordosin增强FK506体外抑制淋巴细胞IL-2分泌作用的研究

目的：观察中药有效成分Nordosin对FK506体外抑制致炎细胞因子作用的影响,探讨Nordosin在移植免疫领域存在的潜在应用价值。方法：建立ConA激活小鼠脾淋巴细胞增殖模型,采用ELISA和RT-PCR方法,测定Nordosin与FK506联合应用对淋巴细胞的IL-2蛋白和mRNA表达的影响;同时,运用流式细胞检测T淋巴细胞CD25^＋表达的变化。结果：Nordosin与FK506联合用药可以抑制小鼠脾淋巴细胞的IL-2分泌（P〈0.01）,IL-2 mRNA表达以及T淋巴细胞CD25＋表达也显著降低（P〈0.01）,其作用强于单独使用FK506组（P〈0.01）。结论：Nordosin可以增强FK506抑制体外淋巴细胞IL-2分泌及其受体表达的作用,Nordosin可望成为一种免疫抑制辅助用药。     

乙双吗啉对有丝分裂原诱导的小鼠淋巴细胞增殖反应的抑制作用

当浓度为1.56mg/L时,乙双吗啉就可明显抑制LPS诱导的体外小鼠淋巴细胞增殖反应,其作用随浓度的增高而加强。当此药浓度增大到6.25mg/L时,ConA和PHA诱导的体外小鼠淋巴细胞增殖反应才开始出现明显抑制。表明乙双吗啉对B细胞增殖反应的抑制作用强于对T细胞增殖反应。乙双吗啉对淋巴细胞无非特异细胞毒作用。     

生长抑素增强胆囊癌细胞化疗敏感性的研究

目的 :探讨在生长抑素预先作用下 ,胆囊癌细胞化疗敏感性的变化。方法 :运用生长抑素预先作用于体外培养的人胆囊癌细胞株 ,2 4h后加入梯度浓度的化疗药物阿霉素 ,观察癌细胞的生长曲线 ,并与单独使用化疗药物的癌细胞生长曲线相比较。结果 :阿霉素能够明显抑制胆囊癌细胞的生长 ,其作用呈现效应 -浓度依赖性 (P <0 .0 5)。生长抑素能够诱导离体胆囊癌细胞生长停滞于DNA合成期 (S期)。经生长抑素预处理后 ,阿霉素对癌细胞生长的抑制效果较单独用药为佳 (P <0 .0 1 )。结论 :生长抑素可增强胆囊癌细胞对化疗药物阿霉素的敏感性。     

FK506和Rapamycin：新的免疫应答药理学探针

近两年已证明有强免疫抑制活性的两种大环内酯抗生素FK506和Rapamycin(Rapa)对延长动物器官异体移植物的存活和预防动物自身免疫疾病很有效。它们都抑制T细胞增殖,但作用在T细胞活化的不同阶段。像环孢菌素A(CsA)一样,FK506抑制抗原或     

用5%葡萄糖和0.9%氯化钠注射液配制FK506在玻璃、聚氯乙烯和聚烯烃容器中的稳定性及吸附作用

FK506是从Streptomyces tsukubaensis培养物中分离而得的一种大环内酯化合物,与淋巴细胞培养物混合后抑制淋巴细胞增殖的作用比环胞菌素大500倍,可逆转或防止动物许多器官移植的排异反应。临床研究表明FK506对肝脏移植受体的免疫抑制作用也比环胞菌素更有效,一般术后立即给药0.075mg/kg·d,其剂型为FK506与水合聚氧乙烯蓖麻油(HCO-60,非离子表面活性剂)制成10mg/ml溶液,静注时用0.9%氯化钠或5%葡萄糖稀释后,短时(2～4h)或持续用药。     

高浓度他克莫司抑制人骨髓间质干细胞增殖及向成骨细胞分化(英文)

目的探讨他克莫司(FK506)对人骨髓间质干细胞(hBMSCs)增殖及向成骨细胞分化的影响。方法 FK506 0.001~5μmo.lL-1处理hBMSCs细胞中,雌二醇0.01μmol·L-1或咖啡因100μmol·L-1为阳性对照组,作用24 h后用BrdU掺入法检测细胞增殖,在促成骨细胞分化液中作用8 d后用比色法检测碱性磷酸酶(ALP)活性,作用12 d后用邻甲酚酞络合法检测钙沉积量;通过检测磷酸盐释放量间接反映钙调神经磷酸酶(CaN)活性,Western印迹法检测核心结合因子α1亚基(Cbfα1)表达。结果与DMSO对照组相比,FK506 0.001~0.01μmol.L-1促进细胞增殖,但对ALP活性及钙沉积量无影响;FK506 0.5~5μmo·lL-1则呈浓度依赖性地抑制细胞增殖,显著抑制ALP活性及减少钙沉积量(P<0.05)。此外,FK5060.1~5μmo·lL-1呈浓度依赖性地降低CaN活性,与相同浓度FK506呈浓度依赖性地下调Cbfα1的表达效应相一致。结论高浓度FK506可通过CaN/Cbfα1通路抑制hBMSCs增殖及向成骨细胞成骨分化。     

木犀草素对膀胱肿瘤T24细胞的影响

目的研究木犀草素对膀胱肿瘤T24细胞体外增殖的抑制和诱导凋亡作用。方法不同浓度木犀草素作用于指数生长期膀胱肿瘤T24细胞后应用MTS比色法检测细胞增殖的抑制作用。通过DNA凝胶电泳检测细胞的凋亡情况。结果浓度为0.01～0.04mg/mL的木犀草素对体外培养的膀胱肿瘤T24细胞有明显的抑制作用。DNA凝胶电泳结果显示:空白对照组没有出现DNA"梯带";其他4组(除木犀草素0.004mg/mL组)均出现不同程度的DNA"梯带",显示出细胞凋亡的特征。结论木犀草素在浓度为0.01～0.04mg/mL时对膀胱肿瘤T24细胞有通过诱导细胞凋亡的方式抑制细胞生长的作用,可能具有治疗膀胱肿瘤的临床价值。     

苦参碱诱导胰腺癌细胞凋亡的实验研究

目的 探讨苦参碱对胰腺癌细胞SW 1990生长抑制和诱导凋亡作用.方法 将不同浓度的苦参碱作用于培养的SW1990细胞,通过MTT比色法、流式细胞仪和DNA凝胶电泳、电子显微镜技术观察检测细胞凋亡,研究分析其对SW1990细胞的生长抑制和诱导凋亡作用.结果 苦参碱浓度为0.5、1.0、1.5g/L时,细胞生长抑制率分别为14.3%、24.8%、41.6%;流式细胞术测定RD横纹肌肉瘤细胞的凋亡率分别为10.3%、18.3%、27.5%;DNA电泳见DNA"梯形"图谱;电镜显示肿瘤细胞的凋亡形态.结论 苦参碱能抑制SW1990细胞增殖,诱导其凋亡.     

Effects of FK506 and other immunosuppressive anti-rheumatic agents on T cell activation mediated IL-6 and IgM production in vitro

The objective of this study was to investigate the therapeutic potential of FK506 and other immunosuppressive agents for the treatment of rheumatoid arthritis (RA), focusing on the effects on in vitro IL-6 production and IL-6-mediated immune response. We employed an in vitro model producing IL-6 via T cell activation in human PBMC, based on the hypothesis that T cells play a central role in the pathogenesis of RA. FK506 potently inhibited IL-6 production from PBMC stimulated with anti-CD3 and anti-CD28 monoclonal antibody (anti-CD3/CD28). Cyclosporin A (CsA) also inhibited the anti-CD3/CD28 induced IL-6 production but was about 100 times less potent than FK506. Dexamethasone (DEX) inhibited both anti-CD3/CD28 and LPS induced IL-6 production at almost the same concentration. Methotrexate (MTX) did not affect cytokine production. Anti-CD3/CD28 stimulated PBMC culture supernatants were found to enhance IgM production in SKW6.4 cells. The effects of anti-CD3/CD28 stimulated culture supernatants in the presence of agents on IgM production in SKW6.4 cells were investigated. FK506 and CsA led to suppression of IgM production induced by culture supernatants probably via inhibition of IgM inducible cytokine production from PBMC. DEX profoundly enhanced IgM production, although IL-6 production from PBMC was strongly inhibited by the agent. MTX decreased IgM production although it has no inhibitory effect on IL-6 production. The present study suggests that FK506 is the most effective among the four agents for the suppression of IL-6 production and IL-6-mediated autoantibody production in T cell activation related autoimmune diseases such as RA.     

SUPPRESSION OF NITRIC OXIDE PRODUCTION BY CYCLOSPORIN A AND FK506A IN RAT VASCULAR SMOOTH MUSCLE CELLS

1. The effects of the immunosuppressants cyclosporin A (CsA) and FK506 on nitric oxide (NO) synthesis induced by lipopolysaccharide (LPS) or cytokines were examined in rat vascular smooth muscle cells (VSMC) in culture. 2. CsA inhibited by up to 90% the accumulation of nitrite induced by LPS, but FK506 had a weaker effect on nitrite accumulation induced by LPS in these cells. Both CsA and FK506 (at 1 μmol/L) significantly inhibited nitrite production induced by recombinant murine interleukin-1β (rIL-1β). 3. Given their differing potency, it is likely that CsA and FK506 suppress induction of NO synthase through different intracellular mechanisms. This action could contribute to the side effects of CsA therapy.     

FK506 potently inhibits T cell activation induced TNF-alpha and IL-1beta production in vitro by human peripheral blood mononuclear cells

The aim of this study was to elucidate the in vitro inhibitory potency of FK506 on production of the inflammatory cytokines, tumour necrosis factor (TNF)-alpha and interleukin (IL)-1beta, with a view to assessing this immunosuppressive agent as a potential anti-rheumatic drug. We employed an in vitro model which produces TNF-alpha and IL-1beta through T cell activation. Human peripheral blood mononuclear cells (PBMC) were cultured with immobilized anti-CD3/CD28 monoclonal antibody in this model. FK506 inhibited anti-CD3/CD28 induced TNF-alpha and IL-1beta production at concentrations less than 1 ng ml(-1). Flow cytometric analysis of intracellular TNF-alpha and IL-1beta positive cells showed that FK506 potently suppresses inflammatory cytokine production from CD14+ monocytes as well as from T cells. Cyclosporin A (CsA) and dexamethasone (DEX) also inhibited the anti-CD3/CD28 induced cytokine production, but were less potent than FK506. FK506 and CsA, but not DEX, specifically inhibited anti-CD3/CD28 induced inflammatory cytokine production without affecting the lipopolysaccaride (LPS) induced effect. Methotrexate (MTX) was completely inactive for suppressing cytokine production under either condition. Anti-CD3/CD28 stimulated PBMC culture supernatants were found to enhance the expression of adhesion molecules in human vascular endothelial cells. FK506, CsA and DEX led to the suppression of adhesion molecule expression probably by inhibiting cytokine production from PBMC. The inhibitory potency of agents on TNF-alpha and IL-1beta production was compared with cytotoxicity and FK506 was not cytotoxic at concentrations several orders of magnitude greater than those required for cytokine inhibition. These results strongly suggest that FK506 may be most effective to specifically prevent T cell activation mediated inflammatory cytokine production in a clinical setting.     

Selective immunosuppression of prodigiosin 25-C and FK506 in the murine immune system.

The immunosuppressive effects of prodigiosin 25-C were studied in comparison with FK506. Both prodigiosin 25-C and FK506 suppressed T cell proliferation in response to concanavalin A (con A) or phytohemagglutinin (PHA) more significantly than that to lipopolysaccharide. However, prodigiosin 25-C inhibited con A-mediated mitogenic response more strongly than PHA-mediated one. FK506 showed no selectivity among those responses. In addition, when higher concentration of con A was used an inhibitory effect of prodigiosin 25-C became more evident whereas that of FK506 became less evident. Furthermore, prodigiosin 25-C affected neither interleukin-2 (IL-2) production nor IL-2 receptor (IL-2R) and transferrin receptor (TF-R) expression in vitro, though FK506 extensively inhibited IL-2 production and significantly suppressed IL-2R and TF-R expression. When comparing the effects of prodigiosin 25-C and FK506 in vivo by injecting antigens of different nature to a mouse, prodigiosin 25-C selectively inhibited cytotoxic T lymphocyte (CTL) activity induced by an allogenic mastocytoma, P815, without affecting production of antibody against a thymus dependent (TD) antigen, sheep red blood cell (SRBC). On the contrary, FK506 significantly inhibited both CTL induction and the antibody production. When Brucella abortus, a thymus independent (TI) antigen, and SRBC were simultaneously challenged to a mouse, neither prodigiosin 25-C nor FK506 affected antibody production against the TI antigen while the effect on the TD antigen were the same as described above. The present results revealed the unique immunosuppressive property of prodigiosin 25-C which was different from that of FK506.     

Rapamycin, but not FK506 and GPI-1046, increases neurite outgrowth in PC12 cells by inhibiting cell cycle progression

Immunophilin ligands such as rapamycin, FK506 and GPI-1046 have been reported to increase neurite outgrowth in vitro and to have neuroprotective activity in vitro and in vivo. In this study, however, FK506 and GPI-1046 (0.1-1000 nM) had little effect on neurite outgrowth in PC12 cells in either the presence or absence of nerve growth factor. In contrast, rapamycin markedly increased neurite outgrowth in PC12 cells in the presence of a low concentration of nerve growth factor (EC(50)=10 nM). Unlike FK506 and GPI-1046, rapamycin is an inhibitor of cell cycle progression. Other cell cycle inhibitors such as ciclopirox and flavopiridol also increased neurite outgrowth in PC12 cells in the presence of a low concentration of nerve growth factor (EC(50)=250 nM and 100 nM, respectively). The neuroprotective effects of FK506, rapamycin and GPI-1046 were also tested in a rodent model of permanent focal cerebral ischemia. FK506 and rapamycin decreased infarct volume by 40% and 37%, respectively, whereas GPI-1046 was ineffective. These data do not support the previous suggestion that FK506 and GPI-1046 increase neurite outgrowth of PC12 cells in vitro. Rapamycin increases neurite outgrowth of PC12 cells, an effect that can be ascribed to its ability to inhibit cell cycle progression. The neuroprotective effect of FK506 and rapamycin against cerebral ischemia is probably not due to differentiation of neuronal precursors or stimulation of neuronal regeneration.     

肾病综合征患者他克莫司血药浓度监测的回顾性分析与评价

目的：探讨肾病综合征患者他克莫司（FK506）“治疗窗”浓度及影响因素,为肾病综合征患者合理应用他克莫司提供参考。方法：以165例肾病综合征患者为研究对象,统计分析服药剂量、年龄、性别及合并用药对他克莫司血药浓度的影响。结果：在日服药剂量相同的情况下,男性患者血药浓度高于女性患者,有统计学差异（P<0.05）;在日服用剂量不超过2.0 mg情况下,不同年龄组间血药浓度总体有统计学差异（P<0.05）;日服用剂量为2.1~4.0 mg时,并用降低FK506浓度的药物组与并用对FK506浓度无影响的药物组间血药浓度有统计学差异（P<0.05）;并应用Logistic回归模型进行多因素分析,结果显示性别是影响FK506血药浓度的因素。结论：肾病综合征患者FK506的血药浓度个体差异较大,应结合患者的治疗效果及临床表现进行全面分析,及时调整治疗方案,实现个体化用药,使FK506在肾病综合征患者中得到最佳治疗效果。