Çelik Payçu Deniz Gözde, Büyükbaba Boral Özden
Istanbul University Istanbul Faculty of Medicine, Department of Medical Microbiology, Istanbul, Turkey.
Mikrobiyol Bul. 2017 Jan;51(1):52-61. doi: 10.5578/mb.48568.
Toxoplasmosis is a common infection with a complicated treatment process. Azithromycin (AZT) is a macrolide antibiotic that can be effectively used in patients with cerebral and ocular toxoplasmosis and has fewer side effects. Chlorella vulgaris (CV), a single-cell green algae that contains nutrients and has various biological effects. CV extract (CVE) has been shown to have protective effects against infections via immune enhancement by increasing the cytotoxicity of NK cells, IL-12 and IFN-γ levels. The aim of this study was to investigate the protective effects of AZT and CV, individually and in combination, against acute toxoplasmosis in mice, and their effects on NK cell cytotoxixity, IL-12, IFN-γ, and IL-2 levels. Six groups of mice (Balb/c) were formed. With the exception of the healthy control (HC) group, all other groups were infected with 1 ml (11 x 104 trofozoit/ml) Toxoplasma gondii RH strain trophozoites. No further action was performed for infected control (IC) group. After 24 hours from trophozoite infection, CVE was given to CV group, AZT to azithromycin group and CVE + AZT combination to CV + AZT group by oral gavage for 6 days. All of the mice from IC, CV, AZT and CV + AZT groups were sacrified on the 8th day of the infection and serum, peritoneal fluid and spleen samples were collected. Trophozoite count of the groups were determined in all groups except HC group and the average growth inhibition activity was calculated by using the growth inhibition formula. In all groups IL-12, IFN-γ, IL-2 levels were measured with ELISA method and cytotoxicity of the NK cells were measured using Cytotox 96 Non-Radioactive Cytotoxicity Assay. The number of trophozoites were significantly lower in the CV group than the IC group (p< 0.001), and also significantly lower in CV + AZT combination group than the AZT group. According to the growth inhibition calculations CV treatment showed 88.6%, AZT treatment 98.46%, AZT + CV combination treatment 99.4% antiprotozoal activity against T.gondii compared with the IC group. NK cell cytotoxicity in the CV and the combination group were significantly higher than all the other groups (p< 0.001). IL-12 and IFN-γ levels were highest in IC group and the lowest in AZT + CV group. This situation has been linked to the fact that the severity of the infection has fallen considerably. IL-2 levels were significantly higher in CV, CV + AZT groups than in the other groups (p< 0.001). In our study, even CV administration alone caused a significant decline in infection.This may be related to the increased NK cytotoxicity, IL-2, IL-12 and IFN-γ levels. CV + AZT combination seems to be an effective treatment option than AZT alone, particularly in patients who are difficult to treat with common methods or in patients with immunosuppression.
弓形虫病是一种常见感染，治疗过程复杂。阿奇霉素（AZT）是一种大环内酯类抗生素，可有效用于脑型和眼型弓形虫病患者，且副作用较少。小球藻（CV）是一种单细胞绿藻，含有多种营养成分并具有多种生物学效应。小球藻提取物（CVE）已被证明可通过提高自然杀伤细胞（NK细胞）的细胞毒性、白细胞介素-12（IL-12）和干扰素-γ（IFN-γ）水平来增强免疫，从而对感染起到保护作用。本研究旨在探究AZT和CV单独及联合使用对小鼠急性弓形虫病的保护作用，以及它们对NK细胞细胞毒性、IL-12、IFN-γ和白细胞介素-2（IL-2）水平的影响。将小鼠（Balb/c）分为六组。除健康对照组（HC）外，所有其他组均感染1 ml（11×10⁴速殖子/ml）刚地弓形虫RH株滋养体。感染对照组（IC）不做进一步处理。滋养体感染24小时后，通过灌胃法给CV组小鼠给予CVE，给阿奇霉素组小鼠给予AZT，给CV + AZT组小鼠给予CVE + AZT组合，持续6天。在感染第8天，处死IC、CV、AZT和CV + AZT组的所有小鼠，收集血清、腹腔液和脾脏样本。除HC组外，测定所有组的滋养体数量，并使用生长抑制公式计算平均生长抑制活性。采用酶联免疫吸附测定法（ELISA）测定所有组的IL-12、IFN-γ、IL-2水平，使用细胞毒性96非放射性细胞毒性测定法测定NK细胞的细胞毒性。CV组的滋养体数量显著低于IC组（p < 0.001），CV + AZT组合组的滋养体数量也显著低于AZT组。根据生长抑制计算结果，与IC组相比，CV治疗对刚地弓形虫的抗原虫活性为88.6%，AZT治疗为98.46%，AZT + CV联合治疗为99.4%。CV组和联合治疗组的NK细胞细胞毒性显著高于所有其他组（p < 0.001）。IL-12和IFN-γ水平在IC组最高，在AZT + CV组最低。这种情况与感染严重程度大幅下降有关。CV组、CV + AZT组的IL-2水平显著高于其他组（p < 0.001）。在我们的研究中，即使单独给予CV也能使感染显著下降。这可能与NK细胞毒性、IL-2、IL-12和IFN-γ水平升高有关。CV + AZT组合似乎比单独使用AZT是一种更有效的治疗选择，特别是对于那些难以用常规方法治疗的患者或免疫抑制患者。
