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评价系统性念珠菌感染小鼠肾脏和脾脏固有免疫细胞和获得性免疫细胞浸润情况及土木香氯仿萃取物的治疗作用。

Evaluation of intrinsic and acquired immune cells infiltration in kidney and spleen of the mice infected with systemic candidiasis and treated with chloroform fraction of Zataria Multiflora Boiss.

机构信息

Department of Pathobiology, Faculty of Veterinary Specialized Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran.

Department of Cellular and Molecular Biology, Faculty of Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran.

出版信息

Microb Pathog. 2018 Jan;114:444-448. doi: 10.1016/j.micpath.2017.11.032. Epub 2017 Nov 26.

Abstract

BACKGROUND AND AIMS

Despite the use of antifungal drugs, the visceral candidiasis is associated with a high mortality rate. The aims of this study were an evaluation of intrinsic and acquired immune cells infiltration in kidney and spleen of the mice infected with systemic candidiasis and treated with chloroform fraction of Zataria Multiflora Boiss.

MATERIALS AND METHODS

Candida albicans (C. albicans) ATCC10231 clinical standard strain was isolated. C. albicans LD50 was determined. The laboratory animal (BALB/C mouse) infection with the visceral candidiasis was performed. The kidney and spleen tissues were stained with PAS and prepared for confirmation under the microscope. The Zataria Multiflora Boiss (Shiraz thyme) was prepared and the effects on the infected group were assessed. The kidney and spleen mononuclear cells (MNCs) were prepared and the flow cytometry technique was performed for the assessment of Th1, Th17, and Treg cells.

RESULTS

The LD50 and LD totals were 1.5 × 108 and 2 × 108 Yeast/0.1 ml, respectively. In mice which had a drug intervention, including chloroform fraction of Zataria Multiflora Boiss, thymol, carvacrol or fluconazole, fungal purification was greater in the spleen than in the kidney. Among those mice without medication intervention, fungal clearance was higher in the kidney. The highest percentage of TH1 cells was in group 1 and then group 4 and in groups 2 and 3 respectively. Moreover, there was a significant difference between groups 4 and 5 and also 6 and 7. The percentage of TH1 cells in the spleen MNCs was higher than that of the kidney cells, which is the difference between the groups except for group 7. The percentage of TH17 cells in the kidney and spleen of all drug-receiving groups exhibited a significant increase compared to groups 6 and 7. The percentage of Treg cells in the kidney and the spleen only in the extract-receiving group had a significant decrease compared to the non-drug receiving group and the other groups receiving group depicted no significant difference in the percentage of Treg cells.

CONCLUSION

In addition to the direct effect on the fungus proven in vitro, the extract exhibits immunosuppressive effects, and thus can degrade the fungus through this way. The results demonstrated that the fraction of Zataria Multiflora Boiss can be considered as a powerful alternative to C. albicans therapy along with other therapies.

摘要

背景与目的

尽管使用了抗真菌药物,内脏念珠菌病仍与高死亡率相关。本研究的目的是评估系统性念珠菌病感染小鼠的固有和获得性免疫细胞浸润,并评估其用百里香氯仿提取物治疗的效果。

材料与方法

分离临床标准株白念珠菌(C. albicans)ATCC10231,确定 C. albicans 的半数致死量(LD50)。采用 BALB/C 小鼠进行内脏念珠菌病感染的实验室动物模型构建。对肾脏和脾脏组织进行 PAS 染色,并在显微镜下进行确认。制备百里香(Shiraz thyme)并评估其对感染组的影响。制备肾脏和脾脏单核细胞(MNCs)并采用流式细胞术评估 Th1、Th17 和 Treg 细胞。

结果

LD50 和 LD 总量分别为 1.5×108 和 2×108 个酵母菌/0.1ml。在接受药物干预(包括百里香氯仿提取物、麝香草酚、香芹酚或氟康唑)的小鼠中,脾脏中的真菌清除率高于肾脏。在未接受药物干预的小鼠中,肾脏中的真菌清除率更高。TH1 细胞百分比最高的是第 1 组,其次是第 4 组,然后是第 2 组和第 3 组。此外,第 4 组和第 5 组以及第 6 组和第 7 组之间存在显著差异。脾脏 MNCs 中的 TH1 细胞百分比高于肾脏细胞,这是除第 7 组之外的组间差异。接受所有药物治疗的小鼠肾脏和脾脏中的 TH17 细胞百分比与第 6 组和第 7 组相比均显著增加。仅在接受提取物的组中,肾脏和脾脏中的 Treg 细胞百分比与未接受药物的组相比显著降低,而其他接受药物的组的 Treg 细胞百分比没有显著差异。

结论

除了体外证明的对真菌的直接作用外,提取物还具有免疫抑制作用,因此可以通过这种方式降解真菌。结果表明,百里香提取物可作为治疗白念珠菌的有效替代方法,与其他疗法联合使用。

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