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Inhibitory effect of hypericin on intracellular proliferation of Toxoplasma gondii in macrophages |
CUI Jie1,2,3, WANG Qi4, YANG Xiao-di2, SUN Xi-meng5, WANG Rui2, JIANG Hui2, WANG Shou-xiang2, ZHANG Min2, WEI Ming2, XIA Hui2, CHEN Xing-zhi2, SHEN Ji-long1 |
1. Department of Parasitology, School of Basic Medicine, Anhui Medical University, Hefei 230032, China; 2. Department of Microbiology and Parasitology, Anhui Key Laboratory of Infection and Immunity, Bengbu Medical College, Bengbu 233030, China; 3. Microscopic Morphology Integrated Laboratory, Bengbu Medical College, Bengbu 233030, China; 4. Anhui Provincial Institute of Schistosomiasis Control, Hefei 230061, China; 5. Department of Parasitology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China |
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Abstract The suppressive effect of hypericin on the intracellular proliferation of Toxoplasma gondii (T.gondii) tachyzoites in mouse peritoneal macrophages was explored in this study. Macrophages were isolated and infected with T. gondii tachyzoites (type I, RH strain) expressing green fluorescent protein (GFP). There were four groups: the control group (macrophages infected with RH-GFP without treatment, group A) and the other three groups (macrophages infected with RH-GFP treated with different concentrations of hypericin: group B, with 100 μg/mL; group C, with 200 μg/mL; and group D with 400 μg/mL). Tachyzoites were collected after 1 hour, 2 hours, and 3 hours of treatment. The infected macrophages and the free parasites were observed under different concentrations of hypericin in various time intervals by fluorescence microscopy and flow cytometry. The ultrastucture of tachyzoites was also observed by transmission electron microscopy. Compared with the control group, the number and fluorescence intensity of tachyzoites were gradually decreased in a concentration and time dependent manner under the experimental condition. The ratio of free parasites to those within macrophages increased and reached the peak at the dosage of 400 μg/mL on 3 hours after treatment. With the extension of treatment duration, it was found that tachyzoites showed swelling and deformed; apparent gap appeared between cell membrane and matrix; more and bigger vacuoles were formed; the cell membrane became dissolved and disrupt, the inner structure overflowed. It’s suggested that the inhibition of hypericin on intracellular proliferation of T. gondii tachyzoites in macrophages is presented in a concentration and time dependent manner in vitro, meanwhile, a remarkable killing effect on free tachyzoites is also found.
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Received: 10 November 2017
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Fund:Supported by the National Natural Science Foundation of China(No. 81471983), the Project on the Natural Science Foundation of Bengbu Medical College (No. BYKY1653), the University Students’ Innovation and Entrepreneurship Training Programs of Anhui Province (No. 201610367025), the Project on the Scientific Research Foundation of Anhui Provincial Institute of Schistosomiasis Control (No. 1603) |
Corresponding Authors:
Shen Ji-long, Emial: shenjilong53@126.com
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