Gabr Ahmed, Mohamed Amr M, Abou Khalil Nasser S, Sayed Alaa El-Din H
Molecular Biology Research & Studies Institute, Assiut University, Assiut, Egypt.
Department of Clinical Pathology, Faculty of Veterinary Medicine, Assiut University, Assiut, Egypt.
Front Immunol. 2025 Mar 31;16:1566496. doi: 10.3389/fimmu.2025.1566496. eCollection 2025.
Diclofenac (DCF) is a commonly utilized medication in the non-steroidal anti-inflammatory drug category that is released into aquatic systems in significant amounts. Chlorella vulgaris (C. vulgaris) is rich in active phytochemicals known for their haemato-immunological boosting properties.
Our objective was to investigate the haemato-immunological protective properties of Chlorella in mitigating the toxic effects of DCF. Five groups of Clarias gariepinus, each comprising 36 fish, were assigned over a two-week period. The groups were assigned as follows: control group, which received a basal diet only; DCF1 group, which received a basal diet and was exposed to 20 μg/L of DCF; DCF2 group, which received a basal diet and was exposed to 10 mg/L of DCF; and Chlorella +DCF1 and Chlorella+DCF2 groups, which were exposed to the same DCF doses as Groups 2 and 3, respectively, while also being fed a diet containing 25% Chlorella.
Exposure to both doses of DCF significantly decreased erythrocyte count, hemoglobin content, white blood cell count, phagocytic index, and lysozyme activity, while increased eosinophil and neutrophil % in an equipotent manner. The low dose caused a more pronounced reduction in packed cell volume (PCV)% and large lymphocyte% compared to the high dose. A significant decline in platelet count was observed only with the low DCF dose, while the high dose led to a decrease in monocyte%. DCF intoxication led to a dose-related decrease in small lymphocyte% and an increase in erythrocyte morphological alterations and interleukin (IL)-6 levels. The DCF2 group exhibited a higher increase in apoptotic RBCs than the DCF1 group. Intervention with Chlorella alongside the two DCF doses significantly normalized RBC count and eosinophil %, increased PCV% and small lymphocyte%, and decreased erythrocyte abnormalities to an equal extent. Large lymphocyte% in the Chlorella+DCF1 group was successfully restored to normal levels. Phagocytic index and lysozyme activity in the supplemented groups were lower, while IL-6 levels were higher than in the DCF groups. The percentage of apoptotic cells decreased with Chlorella administration, with the Chlorella+DCF1 group showing fewer apoptotic cells than the Chlorella+DCF2 group. Histopathological deterioration and excessive collagen deposition were observed in the spleen of DCF groups, while notable improvements were seen following C. vulgaris supplementation.
These findings suggest that dietary inclusion of C. vulgaris may antagonize the haemato-cytological abnormalities induced by DCF intoxication.
双氯芬酸(DCF)是一种常用的非甾体抗炎药,大量释放到水生系统中。普通小球藻(C. vulgaris)富含具有增强血液免疫特性的活性植物化学物质。
我们的目的是研究小球藻在减轻DCF毒性作用方面的血液免疫保护特性。在两周时间内分配了五组尖吻鲈,每组36条鱼。分组如下:对照组,仅接受基础饲料;DCF1组,接受基础饲料并暴露于20μg/L的DCF;DCF2组,接受基础饲料并暴露于10mg/L的DCF;小球藻+DCF1组和小球藻+DCF2组,分别暴露于与第2组和第3组相同剂量的DCF,同时还喂食含有25%小球藻的饲料。
暴露于两种剂量的DCF均显著降低红细胞计数、血红蛋白含量、白细胞计数、吞噬指数和溶菌酶活性,同时以等效方式增加嗜酸性粒细胞和中性粒细胞百分比。与高剂量相比,低剂量导致血细胞比容(PCV)百分比和大淋巴细胞百分比的降低更为明显。仅低剂量DCF导致血小板计数显著下降,而高剂量导致单核细胞百分比下降。DCF中毒导致小淋巴细胞百分比与剂量相关的下降以及红细胞形态改变和白细胞介素(IL)-6水平升高。DCF2组凋亡红细胞的增加高于DCF1组。与两种DCF剂量同时使用小球藻进行干预可使红细胞计数和嗜酸性粒细胞百分比显著恢复正常,增加PCV百分比和小淋巴细胞百分比,并同等程度地减少红细胞异常。小球藻+DCF1组的大淋巴细胞百分比成功恢复到正常水平。补充组的吞噬指数和溶菌酶活性较低,而IL-6水平高于DCF组。随着小球藻的给药,凋亡细胞百分比降低,小球藻+DCF1组的凋亡细胞比小球藻+DCF2组少。在DCF组的脾脏中观察到组织病理学恶化和过多的胶原沉积,而在补充普通小球藻后有显著改善。
这些发现表明,在饮食中添加普通小球藻可能对抗DCF中毒引起的血液细胞学异常。