复方一枝蒿颗粒体外抗弓形虫RH株速殖子的超微结构观察
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摘要
目的探索复方一枝蒿颗粒体外对弓形虫RH株速殖子超微结构的影响。方法收集弓形虫RH株速殖子感染72 h后的昆明(KM)小鼠腹腔冲洗液,加入复方一枝蒿颗粒水溶液使药物终浓度达到0.8 mg/mL,并以0.04 mg/mL剂量阿奇霉素作为阳性药物对照,同时设立空白对照组。置二氧化碳孵箱作用2 h,分别制备吉瑞氏复合染色涂片和透射电镜样本,比较观察各实验组弓形虫速殖子光镜下形态和透射电镜下超微结构。结果空白对照组虫体符合弓形虫速殖子超微结构正常形态,而阳性药物对照组和复方一枝蒿颗粒作用组均显示着色较浅的异常形态虫体:虫体肿胀变形,两端钝圆。细胞质着色浅且偶见空泡,个别虫体细胞膜似有断裂缺如,胞膜不完整。透射电镜下可见阳性药物对照组和复方一枝蒿组弓形虫速殖子细胞质疏松并出现大量不规则空泡等异常形态,复方一枝蒿颗粒还可使弓形虫速殖子超微结构发生胞膜断裂、破损,核膜破损,致密颗粒增多等改变。结论复方一枝蒿颗粒具有损伤弓形虫速殖子超微结构的直接作用。
Objective To investigate the effect of Compound of Artemisia rupestris granule on ultrastructure of tachyzoites of Toxoplasma gondii RH strain in vitro.Methods The peritoneal lavage fluid of KM mice infected 72 hours with Toxoplasma gondii RH strain tachyzoites was collected. The total concentration of the drug was 0.8 mg/mL after the addition of Compound of Artemisia rupestris granule aqueous solution. Azithromycin at a dose of 0.04 mg/mL was used as the positive drug control. Blank control group was set up simultaneously. Incubation with carbon dioxide incubator was performed for 2 h.Samples of Giemsa-Wright compound stained smear and transmission electron microscope were prepared, and the morphology and ultrastructure of T. gondii tachyzoites in each experimental group were observed under microscope and transmission electron microscope respectively.Results In the blank control group, the parasite conformed to the normal morphology of the ultrastructure of the Toxoplasma gondii tachyzoites. Both the positive drug control group and the Compound of Artemisia rupestris granule effect group showed the swelling of the parasites, abnormal morphology such as irregular vacuoles in the cytoplasm. Compound of Artemisia rupestris granule could also cause the breakage and damage of the membrane ultrastructure of Toxoplasma gondii tachyzoites, nuclear membrane damage, increased dense particles and other changes.Conclusions Compound of Artemisia rupestris granule has a direct damage effect on the ultrastructure of Toxoplasma gondii Tachyzoites.
Objective To investigate the effect of Compound of Artemisia rupestris granule on ultrastructure of tachyzoites of Toxoplasma gondii RH strain in vitro.Methods The peritoneal lavage fluid of KM mice infected 72 hours with Toxoplasma gondii RH strain tachyzoites was collected. The total concentration of the drug was 0.8 mg/mL after the addition of Compound of Artemisia rupestris granule aqueous solution. Azithromycin at a dose of 0.04 mg/mL was used as the positive drug control. Blank control group was set up simultaneously. Incubation with carbon dioxide incubator was performed for 2 h.Samples of Giemsa-Wright compound stained smear and transmission electron microscope were prepared, and the morphology and ultrastructure of T. gondii tachyzoites in each experimental group were observed under microscope and transmission electron microscope respectively.Results In the blank control group, the parasite conformed to the normal morphology of the ultrastructure of the Toxoplasma gondii tachyzoites. Both the positive drug control group and the Compound of Artemisia rupestris granule effect group showed the swelling of the parasites, abnormal morphology such as irregular vacuoles in the cytoplasm. Compound of Artemisia rupestris granule could also cause the breakage and damage of the membrane ultrastructure of Toxoplasma gondii tachyzoites, nuclear membrane damage, increased dense particles and other changes.Conclusions Compound of Artemisia rupestris granule has a direct damage effect on the ultrastructure of Toxoplasma gondii Tachyzoites.
引文
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[2]詹婷婷,米荣生,黄燕,等.典型城市屠宰及销售猪肉中弓形虫的分子检测研究[J],中国动物传染病学报,2016,24(6):30-35.
[3]方险峰.新疆一枝蒿药材的研究[J].新疆化工,2017(Z2):5-9.
[4]王大为,郭晓改,司海洋,等.弓形虫感染治疗方法的研究进展[J].黑龙江畜牧兽医,2016(5):54-57,61.
[5]杨莉.新疆特有药用植物:一支篙栽培技术[J].农业开发与装备,2016(6):142-142.
[6]董慧君,蒋云涛,俞可云,等.维药新疆一枝蒿的研究进展[J].现代中药研究与实践,2013,27(4):73-77.
[7]姜雯,姚华,陈强,等.复方一枝蒿微丸抗甲型H1N1流感病毒的研究[J].中药药理与临床,2015,31(6):135-137.
[8]许律捷,姜雯,庞晓丛,等.复方一枝蒿抗流感有效成分的网络药理学研究[J].药学学报,2017,52(5):745-752.
[9]CHAUDHARY K,DARLING J A,FOHL L M,et al.Purine salvage pathways in the apicomplexan parasite Toxoplasma gondii[J].Journalof Biological Chemistry,2004,279(30):31221-31227.
[10]杨秋林,许丽芳,伍和平.四种纯化弓形虫速殖子方法对虫体RNA影响的比较[J].右江民族医学院学报,2006,28(5):825-826.
[11]张健宁,吴洁,陆方贤,等.阿齐霉素抗弓形虫作用的超微结构研究[J].电子显微学报,2000,19(3):317-318.
[12]于朝云,张保德,宁俊雅,等.中草药抗弓形虫作用机制的研究进展[J].中国血吸虫病防治杂志,2015,27(5):555-557,559.
[13]聂福旭,贾志江,杨红梅,等.抗弓形虫药物的类型和作用机制[J].中国动物传染病学报,2013,21(2):75-79.
[14]杨翠萍,万红娇.青蒿素及其衍生物抗弓形虫的研究进展[J].国际医学寄生虫病杂志,2006,(3):160-162.
[15]尹卫东,高全成,刘向东,等.双氢青蒿素和阿奇霉素对小鼠体内弓形虫速殖子超微结构的影响[J].中国寄生虫学与寄生虫病杂志,2009,27(4):325-327.