LHRH-PE40对荷瘤BALB/c小鼠抗肿瘤效果的实验研究

目的 观察LHRH-PE40对小鼠腹水瘤及实体瘤的治疗效果,为确定药理、毒理学实验剂量提供依据。方法 分别给BALB/c小鼠腹腔内和背部皮下接种骨髓瘤SP2/0细胞,接种后第4d和第9d,分别腹腔注射LHRH-PE40,并设空白对照组,停药后第5d处死,观察两组小鼠腹水和腹腔内肿瘤生长情况,称量瘤重,计算抑瘤率。结果 腹水瘤实验组所有小鼠腹部未见腹水,腹腔未见肿瘤。实体瘤实验组抑瘤率为54％。结论LHRH-PE40可以有效地抑制荷瘤小鼠实体瘤的生长。     

IL-2、15、22基因对CVB3 VP1 DNA疫苗免疫效果的影响

目的观察白细胞介素2(Interleukin2,IL2)、白细胞介素15(Interleukin15,IL15)和白细胞介素22(Interleukin22,IL22)基因对柯萨奇病毒(CoxasckievirusB3,CVB3)VP1DNA疫苗诱导小鼠免疫应答及保护作用的影响。方法取雄性BALBc小鼠,随机分为6组,每组20只,分别肌内注射盐水、pcDNA3、pcDNA3VP1、pcDNA3IL2+pcDNA3VP1、pcDNA3IL15+pcDNA3VP1和pcDNA3IL22+pcDNA3VP1,每周1次,共3次,每次免疫后6d取血清用微量中和试验检测CVB3中和抗体,3次免疫后用800TCID50CVB3感染小鼠,观察小鼠的生存时间和生存率。结果pcDNA3VP1、pcDNA3IL2+pcDNA3VP1和pcDNA3IL15+pcDNA3VP1组均能诱导小鼠产生中和抗体,抗体滴度随免疫次数增加而提高。病毒攻击后,各组的生存率差异无显著意义,pcDNA3IL2+pcDNA3VP1和pcDNA3IL15+pcDNA3VP1组较其他4组生存时间明显延长。pcDNA3IL22+pcDNA3VP1组虽也能诱导小鼠产生中和抗体,但抗体滴度和生存情况无明显变化。结论IL2、IL15基因对CVB3VP1DNA疫苗诱导小鼠产生免疫应答和免疫保护有一定的增强作用,而IL22基因无明显作用。     

当归多糖预防给药对放射损伤小鼠骨髓单个核细胞黏附分子表达及细胞周期的影响

目的探讨当归多糖(Angelica polysaccharide,APS)预防给药对放射损伤小鼠骨髓单个核细胞(Bone marrow mononuclear cells,BMNCs)黏附分子表达及细胞周期的影响。方法将BALB/c小鼠分为4组:正常对照组(不作任何处理)、NS组(经腹腔注射生理盐水)、2mg/kgAPS组(经腹腔注射2mg/kgAPS)和8mg/kgAPS组(经腹腔注射8mg/kgAPS),连续注射7d后,进行照射,并继续给药13d。分别于照射后第7、14天采集各组小鼠外周血,并取骨髓,进行WBC、RBC、PLT及BMNC计数;流式细胞术检测小鼠Sca-1+BMNC表面黏附分子CD44和CD49d的表达及BMNC细胞周期的变化;半定量RT-PCR和Western blot法分别检测小鼠BMNC细胞周期蛋白D(2CyclinD2)mRNA转录水平和蛋白表达水平的变化。结果与正常对照组比较,NS组外周血WBC、RBC、PLT及BMNC数量均明显减少,Sca-1+BMNC表面黏附分子CD44和CD49d的表达明显下降(P<0.05),G0/G1期细胞比例显著增加(P<0.05),CyclinD2 mRNA转录水平和蛋白表达水平均明显降低(P<0.05);2mg/kgAPS组和8mg/kgAPS组能增加外周血各指标及BMNC数量(P<0.05),明显降低第7和14天Sca-1+BMNC表面黏附分子CD44和CD49d的表达水平(P<0.05),降低G0/G1期细胞比例(P<0.05),提高CyclinD2 mRNA的转录水平和蛋白表达水平(P<0.05)。结论 APS预防给药能通过降低放射损伤小鼠Sca-1+BMNC表面黏附分子的表达水平,上调BMNC的CyclinD2 mRNA和蛋白表达来加速BMNCG1期向S期的转换,促进造血恢复。     

氨氯地平及5-氟尿嘧啶联合用药对小鼠肝癌H22细胞的抑制作用

目的探讨氨氯地平(amlodipine)及5-氟尿嘧啶(5-fluorouracil,5-Fu)联合用药对小鼠肝癌H22细胞的抑制作用。方法经不同终浓度的氨氯地平(3.125、6.25、12.25、25和50μmol/L)及不同终浓度的5-Fu(12.5、25、50、100和200μg/ml)单独及联合作用小鼠肝癌H22细胞,24、48、72 h后,采用MTT法检测药物对H22细胞生长的抑制作用;流式细胞术及免疫细胞化学法分别检测药物作用48 h后对H22细胞凋亡及细胞中Bcl-2、Bax表达水平的影响。将H22细胞(1.0×107个/ml)经小鼠右腋皮下注射,0.2 ml/只,于注射第2天,将小鼠分为空白对照组(经腹腔注射0.9%生理盐水,0.2 ml/d)、氨氯地平组[用氨氯地平片剂灌胃,10 mg/(0.1 ml·10 g体重·d)]、5-Fu组[分别于接种后第3、6、9天,经腹腔注射5-Fu溶液,10 mg/(1 ml·10 g体重·d)]及联合用药组(氨氯地平及5-Fu的给药方式与剂量同氨氯地平组及5-Fu组),每组10只(雌雄各半),连续给药10 d,次日断颈处死小鼠,称瘤重,并计算肿瘤生长抑制率。结果氨氯地平组、5-Fu组及联合用药组的细胞生长抑制率均随药物浓度增加逐渐上升,且呈剂量-时间依赖性。联合用药组与氨氯地平组及5-Fu组比较,H22细胞的凋亡率明显升高(P0.05);Bcl-2表达水平明显降低,Bax的表达水平明显升高(P0.05);小鼠体内瘤重明显降低(P0.001),肿瘤生长抑制率明显升高(P0.01)。结论氨氯地平与5-Fu联合用药对体内H22细胞的抑制作用明显强于各单独用药组,具有协同作用,为氨氯地平作为一种化疗增敏剂用于肿瘤的临床治疗提供了实验依据。     

人参多糖对鼻咽癌细胞CNE-2裸鼠移植瘤的放疗增敏作用及其可能的机制

目的探讨人参多糖(ginseng polysaccharide,GPS)对人鼻咽癌细胞CNE-2裸鼠移植瘤的放疗增敏作用及可能的机制。方法取对数生长期的鼻咽癌CNE-2细胞,经裸鼠背部皮下注射,1×107个/(0.2 ml·只),待瘤体最大径为4~6 mm时,取20只模型裸鼠,随机分为4组:对照组(经腹腔注射生理盐水)、放疗组(每次每只裸鼠经肿瘤局部给予5 Gy X射线照射,每3 d照射1次,共3次)、GPS组(经腹腔注射GPS,30 mg/kg,每次0.3 ml,每72 h给药1次,共5次)和GPS联合放疗组(经腹腔注射GPS,30 mg/kg,每次0.3 ml,每72 h给药1次,共5次;末次给药48 h后开始放疗,每只每次经肿瘤局部给予5 Gy X射线照射,每3 d照射1次,共3次)。开始给药3周后处死裸鼠,取瘤体,称重,测量肿瘤的长、短径,计算肿瘤体积及抑制率;HE染色观察移植瘤组织病理学改变;Real-time PCR检测移植瘤组织中β-catenin基因mRNA的转录水平;免疫组织化学和Western blot法检测移植瘤组织中β-catenin蛋白的表达水平。结果与对照组相比,GPS组、放疗组和GPS联合放疗组的肿瘤体积及瘤重均明显下降(P0.05),抑制率分别为29.87%、52.60%和74.68%;镜下观察可见,GPS联合放疗组细胞凋亡明显,多数细胞核完全溶解,细胞结构消失,坏死的肿瘤组织呈现均质红染表现;GPS联合放疗组移植瘤中β-catenin mRNA及蛋白的表达水平均显著下降(P均0.05)。结论 GPS可抑制CNE-2细胞在裸鼠体内的生长,且对鼻咽癌的放疗具有增敏作用,其机制可能与抑制β-catenin的表达有关。     

缬氨酸薄荷酯的制备及其生物活性

以N-boc-L-缬氨酸和薄荷为原料,以二环己基碳二亚胺(DCC)及4-二甲氨基吡啶(DMAP)为催化剂合成了缬氨酸薄荷酯(BHX),利用1HNMR、MS及FTIR对产物进行表征。利用电休克惊厥实验及抑制HeLa细胞增殖实验分别对BHX的抗癫痫及体外抑制肿瘤活性进行了初步研究。在最大电休克惊厥实验(MES)中,小鼠腹腔注射给药,当BHX给药量为100 mg/kg时,与生理盐水组及卡马西平组相比,4只预筛选过的小鼠,在0.5、1、2、3、4 h后出现的惊厥数分别为2、2、1、0、0只,显示出与对照品卡马西平类似的抗癫痫活性;在体外抗肿瘤活性实验中,与对照组相比,抑制率最高达到56.5%,显示出明显的抑制肿瘤细胞增殖能力。     

聚乙二醇修饰猪胰高血糖素样肽－2对试验性结肠炎小鼠肠道紧密连接蛋白和炎性因子基因表达的影响 2014年8月5日

本试验旨在研究聚乙二醇（ PEG）修饰猪胰高血糖素样肽－2（ pGLP?2）对结肠炎小鼠肠道紧密连接蛋白和炎性因子基因表达的影响。试验选取24只BALB／C小鼠,随机分为4组,葡聚糖硫酸钠（ DSS）组小鼠饮用3％ DSS 建立小鼠结肠炎模型,DSS＋pGLP?2组和 DSS＋PEG?pGLP?2组饮用3％ DSS且在试验第8天腹腔分别注射pGLP?2和PEG?pGLP?2,饮水组小鼠正常饮水。试验期10 d。结果表明：与饮水组相比, DSS 组小鼠结肠紧密连接闭锁小带基因（ZO?1）的mRNA相对表达量极显著降低（P＜0．01）;与 DSS 组相比,注射 pGLP?2对 ZO?1的mRNA相对表达量没有改善（ P＞0．05）,而注射PEG?pGLP?2可极显著增加ZO?1的mRNA相对表达量（P＜0．01）。与饮水组相比,DSS 组小鼠结肠白细胞介素－6（IL?6）、白细胞介素－10（IL?10）和干扰素－γ基因（INF?γ）的mRNA相对表达量极显著增加（P＜0．01）;与DSS组相比,注射pGLP?2和 PEG?pGLP?2可极显著降低 IL?6、IL?10和 IFN?γ的 mRNA 相对表达量（ P＜0．01）。结果提示,PEG?pGLP?2通过增加肠道紧密连接蛋白的表达、降低结肠炎小鼠炎性细胞因子的表达抑制其炎性病变,且作用效果优于pGLP?2。     

缬氨酸薄荷酯制备及生物活性研究

以L-缬氨酸和薄荷为原料,以二环己基碳二亚胺(DCC)及4-二甲氨基吡啶(DMAP)为催化剂合成了缬氨酸薄荷酯,利用核磁、质谱及红外对产物进行表征。利用电休克惊厥实验及抑制HeLa细胞增殖实验分别对缬氨酸薄荷酯的抗癫痫及体外抑制肿瘤活性进行了初步研究。在最大电休克惊厥实验中,小鼠腹腔注射给药,当缬氨酸薄荷酯给药量为100mg/kg时,与生理盐水组及卡马西平组相比, 4只预筛选过的小鼠,在0.5 、1、2、3、4 h后出现的惊厥数分别为2、2、1、0、0只,显示出了与对照品卡巴列平类似的抗癫痫活性;在体外抗肿瘤活性实验中,与对照组相比,抑制率最高达到52.5%,显示出了明显的抑制肿瘤细胞增殖的能力。     

小分子阳离子六肽免疫调节活性及其对小鼠感染模型的治疗作用

目的分析小分子阳离子六肽(short cationic hexapeptides,SP6)免疫调节活性及其对小鼠感染模型的治疗作用。方法经小鼠腹腔分别注射不同剂量SP6(25、5、1μg/只),并设生理盐水对照组,于注射后0.5、1、2、4和7 h对腹腔细胞进行细胞计数;于注射SP6后1 h,经小鼠腹腔给予10%墨汁,0.2 ml/只,收集注射墨汁后0.5、1、2、4和7 h小鼠腹腔细胞,观察吞噬细胞的吞噬作用;于注射SP6后1 h,采血并分离血清,ELISA法检测趋化因子和细胞因子水平。将大肠埃希菌K88、葡萄球菌及巴氏杆菌,分别经小鼠腹腔注射,0.2 ml/只,3 h后,经小鼠皮下注射SP6溶液(25、5、1μg/只),设生理盐水对照组,观察并记录小鼠接种后精神状态及死亡情况。结果 SP6各剂量组小鼠腹腔细胞数量一直高于对照组,直至注射后7 h。SP6各剂量组小鼠腹腔吞噬细胞在0.5 h前开始活化,0.5 h时观察到吞噬细胞包围、吞噬墨汁颗粒,较对照组吞噬细胞活化时间缩短,吞噬能力增强。SP6注射后1 h,中、低剂量组小鼠血清中鼠巨噬细胞来源趋化因子(macrophage derived chemokine,MDC)、淋巴细胞趋化因子(lymphocyte chemokine,LC)、嗜酸性粒细胞趋化因子(eosinophil chemotactic factor,ECF)、趋化因子(chemotactic factor,CF)、IL-1β、IL-4和TNF-α水平与对照组相当或高于对照组(P0.05);高剂量组小鼠血清中MDC、LC、ECF、CF、IL-1β、IL-4和TNF-α水平均高于对照组,其中LC、IL-1β、TNF-α水平差异有统计学意义(P0.05)。除巴氏杆菌外,经SP6治疗,高、中、低剂量组小鼠存活数均高于对照组。结论 SP6可调节小鼠免疫反应,对小鼠感染模型有治疗作用。     

日本血吸虫重组半胱氨酸蛋白酶抑制剂对小鼠心肌梗死预后的影响及其免疫调节机制

目的 探讨日本血吸虫重组半胱氨酸蛋白酶抑制剂(rSjcystatin)对小鼠心肌梗死(myocardial infarction,MI)预后的影响及其免疫调节机制.方法 建立小鼠MI模型,分别于术后1、3、5、7、14、28 d经腹腔注射PBS及10、25 μg rSjcystatin.术后28 d,记录MI小鼠的生存...     

冻存H_(22)细胞在卡介苗抑瘤小鼠模型中的应用

目的探讨冻存H22肝癌细胞应用于卡介苗抑瘤小鼠模型的可行性。方法制备冻存H22细胞,分别将冻存10和83 d的H22细胞复苏,计算细胞存活率;将高(7.5×106个/ml)、中(5.0×106个/ml)、低(2.5×106个/ml)剂量的新复苏的H22细胞与1 mg/ml治疗用卡介苗混合后经左侧背部皮下免疫BALB/c小鼠,0.1 ml/只,对照组只接种相应浓度的H22细胞,接种后30 d,称量小鼠皮下瘤体重量,并计算试验组的抑瘤率,试验重复2次。结果冻存10和83 d的H22细胞复苏后的存活率分别为85.45%和82.25%。所有对照组小鼠接种H22细胞后均有肿瘤形成,成瘤率均为100%;同一次试验中,各试验组瘤体重量均明显小于同剂量对照组(P<0.05);同一次试验中,各试验组间瘤体重量差异无统计学意义(P>0.05);不同次试验中,同剂量对照组间瘤体重量差异均有统计学意义(P<0.05);各试验组的抑瘤率均大于60%。结论冻存H22细胞可满足抑瘤试验需要,但试验稳定性仍需改进。     

橙皮苷对小鼠急性肝衰竭的保护作用及其机制

目的观察橙皮苷对脂多糖(Lipopolysaccharide,LPS)联合D-氨基半乳糖(D-Galactosamine,D-GalN)所致小鼠急性肝衰竭(Acute hepatic failure,AHF)的保护作用及其机制。方法小鼠腹腔注射LPS(50μg/kg)和D-GalN(800 mg/kg),复制急性肝衰竭模型,分别用橙皮苷(200 mg/kg)或橙皮苷(200 mg/kg)联合锌原卟啉IX(Zinc protoporphyrin IX,ZnPP)(45 mg/kg)干预。造模后6 h检测肝损伤程度和炎症反应强度,48 h统计小鼠死亡率。结果橙皮苷使AHF小鼠血清转氨酶(ALT和AST)水平下降,肝损伤减轻,生存率提高;血清白细胞介素-10(Interleukin-10,IL-10)水平和肝内血红素加氧酶-1(Heme oxygenase-1,HO-1)的表达较AHF小鼠明显增加,肝内HO-1酶活性明显增高;同时,使血清肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)水平、肝组织中Caspase-3蛋白酶和髓过氧物酶(Myeloperoxidase,MPO)活性较AHF小鼠明显降低。ZnPP不影响橙皮苷促进肝内HO-1酶表达,但通过抑制HO-1酶活性,使橙皮苷抗炎和肝损伤保护作用显著降低。结论橙皮苷主要通过诱导HO-1蛋白表达,使HO-1酶活性增强,从而使炎症反应和肝损伤减轻,对LPS联合D-GalN诱导的AHF产生保护作用。     

Ameliorative Effects of 5-Hydroxymethyl-2-furfural (5-HMF) from Schisandra chinensis on Alcoholic Liver Oxidative Injury in Mice

The aim of this paper is to evaluate the protective effect of 5-hydroxymethyl-2-furfural (5-HMF) on acute alcohol-induced liver oxidative injury in mice. 5-HMF, a maillard reaction product, was isolated from the fruits of Schisandra chinensis for animal experiments. Experimental ICR mice were pretreated with different doses of 5-HMF (7.5, 15, and 30 mg/kg) for seven days by gavage feeding. Biochemical markers and enzymatic antioxidants from serum and liver tissue were examined. Our results showed that the activities of ALT (alanine aminotransferase), AST (aspartate transaminase), TC (total cholesterol), TG (triglyceride), L-DLC (low density lipoprotein) in serum and the levels of MDA (malondialdehyde) in liver tissue, decreased significantly (p < 0.05) in the 5-HMF-treated group compared with the alcohol group. On the contrary, enzymatic antioxidants CAT (catalase), GSH-Px (glutathione peroxidase), and GSH SOD (superoxide dismutase) were markedly elevated in liver tissue treated with 5-HMF (p < 0.05). Furthermore, the hepatic levels of pro-inflammatory response marker tumor necrosis factor-alpha (TNF-α) and interleukin-1β (IL-1β) were significantly suppressed (p < 0.05). Histopathological examination revealed that 5-HMF (30 mg/kg) pretreatment noticeably prevented alcohol-induced hepatocyte apoptosis and fatty degeneration. It is suggested that the hepatoprotective effects exhibited by 5-HMF on alcohol-induced liver oxidative injury may be due to its potent antioxidant properties.     

表达重组人可溶性TRAIL的毕氏酵母发酵工艺

目的优化rhsTRAIL毕氏酵母工程菌在5L发酵罐中的发酵表达工艺参数。方法研究培养基种类、细胞密度、甘油含量、pH值、甲醇浓度、甲醇流加速率及诱导时间等参数对工程菌生长及目的蛋白表达的影响,并用电镜观察其诱导肿瘤细胞凋亡特征。结果在无机盐培养基中,按10%接种A600=8～12的种菌后,24h酵母菌增殖至A600=76;甘油含量为1%时,菌体生长速度快(A600=160);培养基pH值5.0,甲醇终浓度为1%时,诱导96h表达量最高达到120mg/L,并具有诱导肿瘤细胞凋亡特征。结论rhsTRAIL毕氏酵母的发酵工艺参数为无机盐培养基,pH值5.0,接种10%A600=8～12的种菌,甘油含量1%,甲醇终浓度1%,诱导96h。     

Effects of Thrombomodulin in Reducing Lethality and Suppressing Neutrophil Extracellular Trap Formation in the Lungs and Liver in a Lipopolysaccharide-Induced Murine Septic Shock Model

Neutrophil extracellular trap (NET) formation, an innate immune system response, is associated with thrombogenesis and vascular endothelial injury. Circulatory disorders due to microvascular thrombogenesis are one of the principal causes of organ damage. NET formation in organs contributes to the exacerbation of sepsis, which is defined as a life-threatening organ dysfunction caused by a dysregulated host response to infection. We have previously reported that recombinant human soluble thrombomodulin (rTM) reduces lipopolysaccharide (LPS)-induced NET formation in vitro. Here, we aimed to show that thrombomodulin (TM)-mediated suppression of NET formation protects against organ damage in sepsis. Mice were injected intraperitoneally (i.p.) with 10 mg/kg LPS. rTM (6 mg/kg/day) or saline was administered i.p. 1 h after LPS injection. In the LPS-induced murine septic shock model, extracellular histones, which are components of NETs, were observed in the liver and lungs. In addition, the serum cytokine (interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), macrophage chemotactic protein-1 (MCP-1), and interleukin-10 (IL-10)) levels were increased. The administration of rTM in this model prevented NET formation in the organs and suppressed the increase in the levels of all cytokines except IL-1β. Furthermore, the survival rate improved. We provide a novel role of TM in treating inflammation and NETs in organs during sepsis.     

Improved stability and tumor targeting of 5‐fluorouracil by conjugation with hyaluronan

To circumvent the rapid clearance in vivo and consequent low tumor targeting of 5‐fluorouracil (5‐Fu), hyaluronan‐5‐fluorouracil conjugate (HA‐5‐Fu) was firstly synthesized and characterized. The stability of HA‐5‐Fu in vitro was evaluated by incubation with phosphate buffered saline, hyaluronidase solution, and mice plasma, respectively. The tumor targeting was tested by in vitro cytotoxicity evaluation and in vivo pharmacokinetics study in plasma and tumor. HA‐5‐Fu with drug loading of 87.674 mg/g was successfully obtained and confirmed. HA‐5‐Fu showed high stability in acidic environment and moderate stability under enzymatic cleavage. The enhanced cytotoxicity of HA‐5‐Fu over 5‐Fu depended on drug concentration, incubation time, and cell lines type. The t_1/2 of HA‐5‐Fu in plasma after injection of prodrug was extended up to 10 times compared with that of 5‐Fu. Notably, AUC_0–t in tumors of HA‐5‐Fu was 3.6 times higher than 5‐Fu, demonstrating its excellent tumor targeting and quite promising prospect in anti‐cancer therapy. © 2013 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 130: 927‐932, 2013     

Targeting Tumor Acidosis and Regulatory T Cells Unmasks Anti-Metastatic Potential of Local Tumor Ablation in Triple-Negative Breast Cancer

Triple-negative breast cancer (TNBC) is an immunologically heterogenous disease that lacks clinically actionable targets and is more likely to progress to metastatic disease than other types of breast cancer. Tumor ablation has been used to increase response rates to checkpoint inhibitors, which remain low for TNBC patients. We hypothesized that tumor ablation could produce an anti-tumor response without using checkpoint inhibitors if immunosuppression (i.e., Tregs, tumor acidosis) was subdued. Tumors were primed with sodium bicarbonate (200 mM p.o.) to reduce tumor acidosis and low-dose cyclophosphamide (100–200 mg/kg i.p.) to deplete regulatory T cells, as has been shown independently in previous studies. A novel injectable ablative was then used to necrose the tumor, release tumor antigens, and initiate an immune event that could create an abscopal effect. This combination of bicarbonate, cyclophosphamide, and ablation, called “BiCyclA”, was tested in three syngeneic models of TNBC: E0771 (C57BL/6), 67NR (BALB/c), and 4T1-Luc (BALB/c). In E0771 and 67NR, BiCyclA therapy significantly reduced tumor growth and cured 5/7 and 6/10 mice 50 days after treatment respectively. In the metastatic 4T1-Luc tumors, for which surgery and checkpoint inhibitors fail, BiCyclA cured 5/10 mice of primary tumors and lung metastases. Notably, CD4+ and CD8+ T cells were found to be crucial for the anti-metastatic response, and cured mice were able to resist tumor rechallenge, suggesting production of immune memory. Reduction of tumor acidity and regulatory T cells with ablation is a simple yet effective therapy for local and systemic tumor control with broad applicability as it is not limited by expensive supplies.     

Captopril Combined with Furosemide or Hydrochlorothiazide Affects Macrophage Functions in Mouse Contact Hypersensitivity Response

Hypertension is a chronic disease associated with chronic inflammation involving activated macrophages. Antihypertensive drugs (for example, angiotensin-converting enzyme inhibitors—ACEIs) used in the treatment of hypertension have immunomodulatory properties. On the other hand, the immunological effect of diuretics and combined drugs (diuretics + ACEI) is unclear. Therefore, we examined the influence of diuretics and combination drugs (ACEI + diuretic) on cellular response (contact hypersensitivity), production of reactive oxygen intermediates (ROIs), and nitric oxide (NO), and the secretion of interleukin-12 (IL-12). CBA mice were administered i.p. captopril (5 mg/kg) with or without hydrochlorothiazide (10 mg/kg) or furosemide (5 mg/kg) for 8 days. On the third day, the mice were administered i.p. mineral oil, and macrophages were collected 5 days later. In the presented results, we show that diuretics administered alone or with captopril increase the generation of ROIs and reduce the formation of NO by macrophages. Moreover, tested drugs inhibit the secretion of IL-12. Diuretics and combined drugs reduce the activity of contact hypersensitivity (both activation and induction phases). Our research shows that the tested drugs modulate the cellular response by influencing the function of macrophages, which is important in assessing the safety of antihypertensive therapy.     

Exploring the Biomaterial-Induced Secretome: Physical Bone Substitute Characteristics Influence the Cytokine Expression of Macrophages

In addition to their chemical composition various physical properties of synthetic bone substitute materials have been shown to influence their regenerative potential and to influence the expression of cytokines produced by monocytes, the key cell-type responsible for tissue reaction to biomaterials in vivo. In the present study both the regenerative potential and the inflammatory response to five bone substitute materials all based on β-tricalcium phosphate (β-TCP), but which differed in their physical characteristics (i.e., granule size, granule shape and porosity) were analyzed for their effects on monocyte cytokine expression. To determine the effects of the physical characteristics of the different materials, the proliferation of primary human osteoblasts growing on the materials was analyzed. To determine the immunogenic effects of the different materials on human peripheral blood monocytes, cells cultured on the materials were evaluated for the expression of 14 pro- and anti-inflammatory cytokines, i.e., IL-6, IL-10, IL-1β, VEGF, RANTES, IL-12p40, I-CAM, IL-4, V-CAM, TNF-α, GM-CSF, MIP-1α, Il-8 and MCP-1 using a Bio-Plex^® Multiplex System. The granular shape of bone substitutes showed a significant influence on the osteoblast proliferation. Moreover, smaller pore sizes, round granular shape and larger granule size increased the expression of GM-CSF, RANTES, IL-10 and IL-12 by monocytes, while polygonal shape and the larger pore sizes increased the expression of V-CAM. The physical characteristics of a bone biomaterial can influence the proliferation rate of osteoblasts and has an influence on the cytokine gene expression of monocytes in vitro. These results indicate that the physical structure of a biomaterial has a significant effect of how cells interact with the material. Thus, specific characteristics of a material may strongly affect the regenerative potential in vivo.