他克莫司的药物不良反应

他克莫司(Tacrolimus，FK506)的药物不良反应较少，主要为肾毒性，也可见神经毒性、胃肠道反应等。现将该药在国内报道的药物不良反应综述如下。     

肝移植后他克莫司血药浓度对外周血单个核细胞内HBVDNA的影响

背景:肝移植后他克莫司等免疫抑制剂的长期应用导致机体细胞免疫功能降低,并有可能影响机体对乙肝病毒的清除.目的:分析乙肝相关肝移植患者后不同浓度他克莫司对外周血单个核细胞中的HBVDNA含量的影响.方法:纳入乙肝相关终末期肝病肝移植受者23例,根据移植后12周清晨空腹他克莫司血药浓度,分为高浓度组(≥10 μg/L)9 例和低浓度组(<10 μg/L)14例,同时用荧光标记单克隆抗体结合流式细胞技术检测外周血T细胞亚群的百分比,用实时荧光定量PCR检测外周血单个核细胞内的HBV DNA.结果与结论:用多元线性回归分析外周血单个核细胞内的HBV DNA含量与CD8+CD152+呈正相关,与CD8+CD28+呈负相关.高血药浓度他克莫司的患者外周血单个核细胞内的HBV DNA高于低浓度组,其改变与反映细胞免疫功能的指标CD8+CD152+和CD8+CD28+的变化有关.     

肝移植后他克莫司血药浓度对外周血单个核细胞内HBV DNA的影响

背景：肝移植后他克莫司等免疫抑制剂的长期应用导致机体细胞免疫功能降低,并有可能影响机体对乙肝病毒的清除。目的：分析乙肝相关肝移植患者后不同浓度他克莫司对外周血单个核细胞中的HBVDNA含量的影响。方法：纳入乙肝相关终末期肝病肝移植受者23例,根据移植后12周清晨空腹他克莫司血药浓度,分为高浓度组（≥10μg/L）9例和低浓度组（〈10μg/L）14例,同时用荧光标记单克隆抗体结合流式细胞技术检测外周血T细胞亚群的百分比,用实时荧光定量PCR检测外周血单个核细胞内的HBVDNA。结果与结论：用多元线性回归分析外周血单个核细胞内的HBVDNA含量与CD8＋CD152＋呈正相关,与CD8＋CD28＋呈负相关。高血药浓度他克莫司的患者外周血单个核细胞内的HBVDNA高于低浓度组,其改变与反映细胞免疫功能的指标CD8＋CD152＋和CD8＋CD28＋的变化有关。     

肾移植患者服用他克莫司的药物基因组学因素分析

目的探讨CYP3A5基因多态性对服用他克莫司的肾移植患者的药代动力学参数的影响.方法选择24例肾移植患者,治疗方案为他克莫司+霉酚酸酯+强的松联合用药;CYP3A5基因分型采用PCR法和Pyrosequencing测序法;药代动力学模型采用LR法和GLM法.分析肾移植患者性别、CYP3A5×1等位基因、肾移植时年龄、肝肾功能对他克莫司药代动力学的影响.结果基因型CYP3A5×3/×317例,CYP3A5×1/×36例,CYP3A5×1/×11例.肾移植后14d及1、3、6、12个月时,CYP3A5*1等位基因与较低的他克莫司剂量调整浓度相关(P值为0.000);移植术后3个月,肝肾功能与他克莫司的剂量调整浓度相关(P值为0.028).性别因素对他克莫司药代动力学无明显影响.结论在达到相同目标浓度的情况下,与携带CYP3A5×1基因型的个体相比,CYP3A5×3基因型需要更低的用药剂量.     

不同血药浓度他克莫司对肾病综合征的疗效影响分析

目的分析不同血药浓度他克莫司对肾病综合征的疗效影响。方法选取本院2012年5月—2016年5月收治的90例肾病综合征患者为研究对象,患者服用他克莫司达到稳态血药浓度后,应用酶联免疫吸附测定法测定他克莫司全血谷浓度。随访3个月,观察药物疗效,比较患者给药剂量及血药浓度,分析患者不良反应发生率。结果治疗后完全缓解76例(84.4%),部分缓解9例(10.0%),无效5例(5.6%);完全缓解组、部分缓解组、无效组给药剂量分别为(0.05±0.01)mg/(kg·d)、(0.06±0.01)mg/(kg·d)、(0.07±0.02)mg/(kg·d),血药浓度为(8.44±3.25)ng/m L、(5.92±1.53)ng/m L、(3.34±0.86)ng/m L;完全缓解组、部分缓解组不良反应发生率显著高于无效组(P0.05)。结论不同血药浓度他克莫司对肾病综合征的疗效有影响,临床上应将他克莫司全血谷浓度控制在5.19~11.69 ng/m L内,疗效满意。     

他克莫司血药浓度与临床疗效及不良反应关系的研究

目的探讨肾移植患者他克莫司（FK506）血药浓度与效应关系。方法采用微粒子酶免疫法测定56例肾移植术后患者口服FK506后12h的全血药浓度，对患者随访，观察排斥反应的发生及药物的肾毒性，并进行回顾性分析。结果肾移植术后FK506的适宜治疗浓度范围0～1个月为9～14ng／ml，2～3个月为8—12ng／ml，4～6个月为6～10ng／ml，〉7个月为4～6ng／ml。术后发生排斥反应3例，药物肝毒性2例，血糖升高10例。经减药治疗后仍有4例需要胰岛素控制血糖。术后发生感染5例。结论FK506具有良好的免疫抑制效果，其在上述治疗窗范围内，既能达到满意的免疫抑制效果，又能减少排斥反应和肾毒性反应的发生。     

肾移植删艮用他克莫司的药物基因组学因素分析

目的探讨CYP3A5基因多态性对服用他克莫司的肾移植患者的药代动力学参数的影响。方法选择24例肾移植患者,治疗方案为他克莫司＋霉酚酸酯＋强的松联合用药;CYP3A5基因分型采用PCR法和Pyrosequencing测序法;药代动力学模型采用LR法和GLM法。分析肾移植患者性别、CYP3A5×1等位基因、肾移植时年龄、肝肾功能对他克莫司药代动力学的影响。结果基因型CYP3A5×3／×317例,CYP3A5×1／×36例,CYP3A5×1／×11例。肾移植后14d及1、3、6、12个月时,CYP3A5＊1等位基因与较低的他克莫司剂量调整浓度相关（P值为0．000）;移植术后3个月,肝肾功能与他克莫司的剂量调整浓度相关（P值为0．028）。性别因素对他克莫司药代动力学无明显影响。结论在达到相同目标浓度的情况下,与携带CYP3A5×1基因型的个体相比,CYP3A5×3基因型需要更低的用药剂量。     

应用他克莫司出现癫痫1例

患者,女,28岁,因急性混合细胞性白血病17个月,骨髓移植术后1年入院,该患者染色体正常,经米托蒽醌＋长春地辛＋阿糖胞苷＋泼尼松（MOAP）方案达完全缓解（CR）,后再MOAP方案巩固1疗程,因其人类白细胞抗原（HLA）配型为HLAA26,31;B51,58;DR14,17,与其胞姐完全相合,于2004年7月5日行造血干细胞移植,预处理为马法兰／环磷酰胺（BU／CY）方案,为。供A血型不合移植,术后造血重建顺利,术后用环孢紊（CSA）进行移植物抗宿主病（GVHD）治疗,于移植第5个月停用CSA,后出现慢性GVHD（口腔,皮肤,肝脏受累）,于2005年4月10改用他克莫司3mg每12小时1次及甲泼尼龙40mg每日1次控制,半个月前自行减药,出现大便次数增多,大便呈墨绿色,入院体格检查：血压140／85mmHg（1mmHg=0．133kPa）全身皮肤见广泛陈旧性暗红色皮疹,口腔黏膜多处破溃,全身淋巴结无肿大,巩膜无黄染,胸骨无压痛,两肺呼吸音清,心率89次／min,腹平软,肝脾肋下未触及,病理征阴性。     

他克莫司对特发性膜性肾病的治疗观察

目的:研究他克莫司(FK 506)治疗表现为肾病综合征(NS)的特发性膜性肾病的疗效和安全性.方法:17例原发性肾病综合征患者经肾活检确诊膜性肾病,单用激素治疗无效或复发.随机分两组分别予FK 506联用糖皮质激素治疗(治疗组),或环磷酰胺(CTX)联用糖皮质激素治疗(对照组),疗程至少6个月,观察各组的疗效及安全性.结果:治疗组24 h尿蛋白定量、血浆白蛋白、血胆固醇和甘油三酯等指标较对照组均有明显改善,不良反应可以耐受.对照组1例因白细胞减少停止CTX治疗.结论:与CTX相比,FK 506联合激素治疗特发性膜性肾病,短期疗效肯定,不良反应可以耐受.     

他克莫司治疗口腔扁平苔藓的临床效果

目的探讨他克莫司治疗口腔扁平苔藓(OLP)的临床效果。方法选取2015年12月至2018年12月本院收治的61例OLP患者作为研究对象,根据数字奇偶法将其分为对照组(31例,糠酸莫米松乳膏)和观察组(30例,他克莫司软膏)。比较两组的临床疗效、临床体征和症状评分、血清炎症因子水平、不良反应发生情况及复发情况。结果观察组的治疗总有效率明显高于对照组(P<0.05)。治疗后,两组的临床体征、疼痛症状评分均降低,且观察组明显低于对照组(P<0.05)。治疗后,两组的血清IL-6、IL-8、TNF-α水平均降低,且观察组明显低于对照组(P<0.05)。两组的不良反应总发生率比较,差异无统计学意义(P>0.05)。48例患者获得为期12个月的随访,观察组的复发率明显低于对照组(P<0.05)。结论他克莫司治疗OLP的效果显著,可缓解患者的临床体征和疼痛症状,降低血清炎症因子水平,且复发率低。     

HBV携带者肝细胞体外培养的研究

目的:体外培养HBV携带者人原代肝细胞,研究细胞内HBV的复制情况及持续时间,为临床抗病毒试验治疗奠定基础.方法:分离HBV携带者人原代肝细胞进行体外培养.14 d后,测绘细胞的生长曲线和细胞核型分析.分别用巢式PCR和ELISA法对细胞匀浆及培养上清液进行HBV-DNA和HBsAg、HBeAg的检测.结果:HBV携带者人原代肝细胞和培养上清液可检测到HBV-rcDNA,培养上清液内检测到HBsAg和HBeAg.细胞培养3周时开始出现大量死亡.结论:该细胞体外培养期间仍能保持对HBV感染和复制的特性,为体外研究HBV感染情况奠定了基础.     

酒精对乙型肝炎病毒复制和基因表达的影响

目的研究酒精对乙肝病毒复制和基因表达的影响,并探讨其发生机制。方法通过灌胃途径对实验组HBV转基因小鼠予以酒精干预,并在实验第6周末处死。同期设生理盐水组作为对照。定量测定血清HBV-DNA、HBsAg、HBeAg水平,肝组织切片行HBcAg免疫组化。检测血清HBV-DNA阳性查体者戒酒前后血清病毒载量水平。结果HBV转基因小鼠酒精组血清HBV-DNA载量、HBsAg水平、HBeAg阳性率高于对照组;转基因小鼠酒精组肝细胞HB-cAg免疫组化阳性率较对照组明显升高;轻度饮酒组戒酒前后病毒载量差异无统计学意义,中度饮酒组与重度饮酒组戒酒前后病毒载量差异均有统计学意义。结论酒精能促进体内HBV-DNA复制和基因表达。     

Inhibition of replication of hepatitis B virus by cytallene in vitro.

The acyclic cytosine nucleoside analog cytallene [1-(4'-hydroxy-1',2'-butadienyl)cytosine], which has both (+)- and (-)-enantiomers, was evaluated for its anti-hepatitis B virus (HBV) activity in 2.2.15 cells and was found to have potent activity against HBV DNA synthesis. The R-(-)-enantiomer was found to be the more active of the cytallene enantiomers, with a 50% inhibition concentration against HBV synthesis (HBIC50) of 0.08 microM. Its antiviral activity could be reversed by deoxycytidine (dC) and less efficiently by cytidine. Upon removal of the R-(-)-enantiomer from culture medium, the synthesis of HBV DNA could reinitiate, which suggested that the antiviral action is reversible. The R-(-)-enantiomer was also found to be more cytotoxic than the S-(+)-enantiomer. The degree of cytotoxicity varied among the cell lines, with a 50% inhibition of cell growth at greater than 10 microM. The R-(-)-enantiomer had no effect on HBV RNA synthesis and mitochondrial DNA synthesis at a concentration of 10 times or more than the HBIC50. The two enantiomers cannot be deaminated by dC deaminase, and they can be phosphorylated by cytoplasmic dC kinase. The R-(-)-enantiomer of cytallene is the first acyclic cytosine analog with potent inhibitory activity against HBV similar to those of other L-(-)-ddC analogs.     

Efficacies of entecavir against lamivudine-resistant hepatitis B virus replication and recombinant polymerases in vitro

Entecavir (ETV) is a potent and selective inhibitor of hepatitis B virus (HBV) replication in vitro and in vivo that is currently in clinical trials for the treatment of chronic HBV infections. A major limitation of the current HBV antiviral therapy, lamivudine (3TC), is the emergence of drug-resistant HBV in a majority of treated patients due to specific mutations in the nucleotide binding site of HBV DNA polymerase (HBV Pol). To determine the effects of 3TC resistance mutations on inhibition by ETV triphosphate (ETV-TP), a series of in vitro studies were performed. The inhibition of wild-type and 3TC-resistant HBV Pol by ETV-TP was measured using recombinant HBV nucleocapsids, and compared to that of 3TC-TP. These enzyme inhibition studies demonstrated that ETV-TP is a highly potent inhibitor of wild-type HBV Pol and is 100- to 300-fold more potent than 3TC-TP against 3TC-resistant HBV Pol. Cell culture assays were used to gauge the potential for antiviral cross-resistance of 3TC-resistant mutants to ETV. Results demonstrated that ETV inhibited the replication of 3TC-resistant HBV, but 20- to 30-fold higher concentrations were required. To gain further perspective regarding the potential therapeutic use of ETV, its phosphorylation was examined in hepatoma cells treated with extracellular concentrations representative of drug levels in plasma in ETV-treated patients. At these concentrations, intracellular ETV-TP accumulated to levels expected to inhibit the enzyme activity of both wild-type and 3TC-resistant HBV Pol. These findings are predictive of potent antiviral activity of ETV against both wild-type and 3TC-resistant HBV.     

二甲基亚砜促进乙型肝炎病毒对HepG2细胞的吸附作用

目的初步探讨乙型肝炎病毒(HBV)感染经二甲基亚砜(DMSO)处理的HepG2细胞的早期吸附过程,为HBV体外感染机制的研究提供细胞学依据。方法将HepG2细胞分为DMSO处理组和对照组,分别以含有1.5%DMSO和不含DMSO的DMEM培养液培养4d,观察HepG2细胞形态的变化。另将ECV304细胞设为阴性对照组以DMSO处理,3组细胞培养24h后分别以HBV阳性血清孵育2h,收集胰酶消化液及HepG2、ECV304细胞,采用聚合酶链式反应(PCR)分别检测各组的HBV DNA;同时设立空白对照组,采用间接免疫荧光法(IIF)检测乙型肝炎表面抗原(HBsAg)在HepG2细胞上的定位。结果 DMSO处理组的HepG2细胞体积明显增大;DMSO处理组和对照组HepG2细胞内和胰酶消化液中均可检出HBV DNA,但DMSO处理组表达较强;IIF检测结果显示,DMSO处理组的HepG2细胞膜和细胞质的绿色荧光信号明显增强,而阴性对照组的HBV DNA及IIF检测均为阴性。结论 DMSO能在一定程度上促进HBsAg的吸附,从而有助于感染早期过程的实现。     

Effects of human leucocyte interferon on hepatitis B virus replication and immune responses in patients with chronic hepatitis B infection.

Eight patients with chronic hepatitis B infection (seven with chronic active hepatitis and one with chronic persistent hepatitis) were treated with daily intramuscular injections of human leucocyte interferon for periods of 5 to 8 weeks and in one case for 5 months. In one patient there was a marked fall in virus-associated DNA polymerase activity and in the number of DNA containing viral particles during each of two courses of interferon. Hepatitis Be antigen (HBeAg) also disappeared, the aspartate transaminase levels fell and liver histology improved. In the four other patients with detectable DNA polymerase activity there was an early fall but this was transient and in one of these patients there was a continuing rise in activity despite treatment. One other patient became HBeAg negative but hepatitis B surface antigen (HBsAg) titres were mostly unaffected by treatment. A marked decrease in T-lymphocyte mediated cytotoxicity towards HBsAg coated target cells was demonstrated and raises the possibility that an immunosuppressant action of interferon may offsets its direct anti-viral action but may also account for the improvement in liver function which occurred in some patients.     

Silver nanoparticles inhibit hepatitis B virus replication

BACKGROUND: Silver nanoparticles have been shown to exhibit promising cytoprotective activities towards HIV-infected T-cells; however, the effects of these nanoparticles towards other kinds of viruses remain largely unexplored. The aim of the present study was to investigate the effects of silver nanoparticles on hepatitis B virus (HBV). METHODS: Monodisperse silver nanoparticles with mean particle diameters of approximately 10 nm (Ag10Ns) and approximately 50 nm (Ag50Ns) were prepared from AgNO3 in HEPES buffer. The in vitro anti-HBV activities of these particles were determined using the HepAD38 cell line as infection model. RESULTS: Ag10Ns and Ag50Ns were able to reduce the extracellular HBV DNA formation of HepAD38 cells by >50% compared with the vehicle control (that is, HepAD38 cells in the absence of silver nanoparticles). Silver nanoparticles had little effect on the amount of HBV covalently closed circular DNA (cccDNA), but could inhibit the formation of intracellular HBV RNA. Gel mobility shift assays indicated that Ag10Ns bound HBV double-stranded DNA at a DNA:silver molar ratio of 1:50; an absorption titration assay showed that the nanoparticles have good binding affinity for HBV DNA with a binding constant (Kb) of (8.8 +/- 1.0)x10(5) dm(3)mol(-1). As both the viral and Ag10Ns systems are in the nanometer size range, we found that Ag10Ns could directly interact with the HBV viral particles as revealed by transmission electronic microscopy. CONCLUSIONS: Silver nanoparticles could inhibit the in vitro production of HBV RNA and extracellular virions. We hypothesize that the direct interaction between these nanoparticles and HBV double-stranded DNA or viral particles is responsible for their antiviral mechanism.     

鸭乙型肝炎病毒在鸭肝原代细胞中复制的试验观察

取正常雏鸭肝脏制备原代细胞,在细胞培养板中进行鸭乙型肝炎病毒（ＤＨＢＶ）感染,通过ＰＣＲ、紫外分光光度仪、细胞原位杂交等实验技术的检测,观察ＤＨＢＶ在鸭肝原代细胞中的复制。结果,ＤＨＢＶ感染鸭肝原代细胞后３ｄ,病毒即表现出明显的复制现象,在随后一周内细胞内ＤＨＢＶ复制逐渐增强;第二周病毒复制能力开始逐渐衰减,３５ｄ后复制能力基本消失。病毒分布和复制区域主要在肝细胞核内。     

自噬与乙型肝炎病毒复制的关系

目的：探究自噬与乙型肝炎病毒(hepatitis B virus,HBV)复制之间的相互关系。方法利用 HepG2细胞,转染绿色荧光标记的微管相关蛋白轻链3(GFP-LC3),分别在正常和饥饿条件下培养,另共转染 HBV WT 和GFP-LC3,正常条件培养,免疫荧光观察细胞自噬;分别转染1.3 mer HBV 野生型质粒(HBV WT)与 HBV 突变型(HBV X-)质粒,正常和饥饿两种条件下培养,蛋白印迹检测自噬相关蛋白 LC3和乙型肝炎病毒核心蛋白(HBc)的表达;细胞转染 HBV WT 后,分别正常条件培养(对照组),无血清饥饿培养(饥饿组),加入自噬抑制剂3-甲基腺嘌呤(3-MA)培养(3-MA 组),采用酶联免疫吸附测定法(ELISA)检测细胞上清乙型肝炎表面抗原(HBsAg)和乙型肝炎 e 抗原(HBeAg)的含量。结果转染 HBV WT 质粒的细胞内自噬荧光颗粒较对照组明显增加发生自噬的细胞比例分别为13%和2%;饥饿组 HBsAg 含量(1.856±1.415)较对照组(1.531±0.944)升高,HBeAg 含量(0.533±0.391)较对照组(0.334±0.110)升高,差异具有统计学意义(P <0.05);自噬抑制剂组 HBsAg 含量(1.184±0.759)较饥饿组(1.856±1.415)降低,HBeAg 含量(0.306±0.130)较饥饿组(0.533±0.391)降低(P <0.05);蛋白印迹显示,转染HBV WT 质粒的细胞中 LC3蛋白表达较 HBV X-组增高,且在饥饿条件下,HBc 蛋白的表达增高。结论乙型肝炎病毒可诱导细胞自噬的发生,且自噬可促进乙型肝炎病毒的复制。