国际医学寄生虫病杂志
國際醫學寄生蟲病雜誌
국제의학기생충병잡지
INTERNATIONAL JOURNAL OF MEDICAL PARASITIC DISEASES
2013年
5期
245-250
,共6页
抗菌肽%弓形虫%细胞毒性%MTF比色法%瑞姬氏染色法%超微结构
抗菌肽%弓形蟲%細胞毒性%MTF比色法%瑞姬氏染色法%超微結構
항균태%궁형충%세포독성%MTF비색법%서희씨염색법%초미결구
Antimicrobial peptide%Toxoplasma gondii%Cell toxicity%MTT chromatometry%Giemsa-Wright's staining%Ultrastructure
目的 研究家蝇幼虫抗菌肽对弓形虫的杀伤作用. 方法 通过研磨、高速离心和层析分离家蝇幼虫抗菌肽,暂命名为AMP-1.将AMP-1用PBS调整浓度为12.5、25.0和50.0 μg/ml,分别作用于体外游离的和Vero细胞内的弓形虫速殖子,采用MTT法和瑞姬氏染色法观察其24、48和72h的抑制率,并采用MTT法观察其对宿主细胞的毒性作用.将50 μg/ml的家蝇幼虫抗菌肽作用于体外游离的弓形虫速殖子24h后,采用透射电镜观察弓形虫速殖子超微结构的变化. 结果 研究发现,AMP-1的抗游离弓形虫作用随着浓度的增大和时间的延长而升高(F24h=23.482,F48h=20.519,F72h=9.469,P＜0.05).瑞姬氏染色结果显示,各浓度抗菌肽组弓形虫对Vero细胞的侵入率均随着AMP-1浓度的增大而减小(F24h=8.888,F48h=10.890,P＜0.05).12.5、25.0和50.0 μg/ml AMP-1作用于Vero细胞72 h后,细胞存活率分别为94.04％,90.70％和81.94％.透射电镜观察表明,AMP-1能引起弓形虫速殖子细胞膜、胞内细胞器以及细胞核受损. 结论 AMP-1能有效杀伤弓形虫速殖子,并且对正常细胞损伤很小,提示AMP-1可作为抗弓形虫的候选药物.
目的 研究傢蠅幼蟲抗菌肽對弓形蟲的殺傷作用. 方法 通過研磨、高速離心和層析分離傢蠅幼蟲抗菌肽,暫命名為AMP-1.將AMP-1用PBS調整濃度為12.5、25.0和50.0 μg/ml,分彆作用于體外遊離的和Vero細胞內的弓形蟲速殖子,採用MTT法和瑞姬氏染色法觀察其24、48和72h的抑製率,併採用MTT法觀察其對宿主細胞的毒性作用.將50 μg/ml的傢蠅幼蟲抗菌肽作用于體外遊離的弓形蟲速殖子24h後,採用透射電鏡觀察弓形蟲速殖子超微結構的變化. 結果 研究髮現,AMP-1的抗遊離弓形蟲作用隨著濃度的增大和時間的延長而升高(F24h=23.482,F48h=20.519,F72h=9.469,P＜0.05).瑞姬氏染色結果顯示,各濃度抗菌肽組弓形蟲對Vero細胞的侵入率均隨著AMP-1濃度的增大而減小(F24h=8.888,F48h=10.890,P＜0.05).12.5、25.0和50.0 μg/ml AMP-1作用于Vero細胞72 h後,細胞存活率分彆為94.04％,90.70％和81.94％.透射電鏡觀察錶明,AMP-1能引起弓形蟲速殖子細胞膜、胞內細胞器以及細胞覈受損. 結論 AMP-1能有效殺傷弓形蟲速殖子,併且對正常細胞損傷很小,提示AMP-1可作為抗弓形蟲的候選藥物.
목적 연구가승유충항균태대궁형충적살상작용. 방법 통과연마、고속리심화층석분리가승유충항균태,잠명명위AMP-1.장AMP-1용PBS조정농도위12.5、25.0화50.0 μg/ml,분별작용우체외유리적화Vero세포내적궁형충속식자,채용MTT법화서희씨염색법관찰기24、48화72h적억제솔,병채용MTT법관찰기대숙주세포적독성작용.장50 μg/ml적가승유충항균태작용우체외유리적궁형충속식자24h후,채용투사전경관찰궁형충속식자초미결구적변화. 결과 연구발현,AMP-1적항유리궁형충작용수착농도적증대화시간적연장이승고(F24h=23.482,F48h=20.519,F72h=9.469,P＜0.05).서희씨염색결과현시,각농도항균태조궁형충대Vero세포적침입솔균수착AMP-1농도적증대이감소(F24h=8.888,F48h=10.890,P＜0.05).12.5、25.0화50.0 μg/ml AMP-1작용우Vero세포72 h후,세포존활솔분별위94.04％,90.70％화81.94％.투사전경관찰표명,AMP-1능인기궁형충속식자세포막、포내세포기이급세포핵수손. 결론 AMP-1능유효살상궁형충속식자,병차대정상세포손상흔소,제시AMP-1가작위항궁형충적후선약물.
Objective To investigate the anti-Toxoplasma gondii (T.gondii) activity of antimicrobial peptide (AMP) isolated from housefly larvae (Musca domestica).Methods The antimicrobial peptide with anti-T.gondii activity from housefly larvae was isolated through grinding,high speed centrifugation and chromatography,which was named AMP-1.AMP-1 was adjusted to 12.5,25.0 and 50.0 μg/ml with PBS,added to free and intracellular T.gondii tachyzoites and the inhibition rate of T.gondii was detected after 24,48 and 72 h respectively by MTT chromatometry and Giemsa-Wright's staining.The toxicity of AMP-1 on Vero cells was detected by MTT chromatometry when AMP-1 was adjusted to 12.5,25.0 and 50.0 μg/ml with PBS.And then,the ultrastructure of free T.gondii tachyzoites treated with AMP-1 of 50.0 μg/ml was observed by transmission electron microscope (TEM) after 24 h.Results It was shown that anti-free T.gondii activity of AMP-1 increased along with the increase of concentration of AMP-1 (F24h=23.482,F48h=20.519,F72h=9.469,P＜0.05).The invasion rate of T.gondii for Vero cells decreased with the increase of the concentration of AMP-1 (F24h=8.888,F48h=10.890,P＜0.05).And the survival rate of Vero cells was 94.04％,90.70％ and 81.94％ respectively when treated with AMP-1 of 12.5 μg/ml to 50.0 μg/ml.Through TEM,it was observed that cell membrane,nucleus and cytoplasm of T.gondii tachyzoites were all destroyed.Conclusion AMP-1 can kill T.gondii tachyzoites effectively,and has little damage on normal cells.It implies that AMP-1 may be served as a candidate for anti-Toxoplasma drug.
