摘要
目的观察复方一枝蒿颗粒单用或联合阿奇霉素治疗小鼠急性弓形虫感染效果。方法通过腹腔注射弓形虫RH株速殖子的方法建立KM小鼠急性弓形虫感染动物模型,随机分成4组:急性弓形虫感染模型对照组、复方一枝蒿颗粒治疗组、复方一枝蒿颗粒与阿奇霉素联合治疗组,阿奇霉素对照组。观察小鼠病症及存活情况,采用ELISA法检测小鼠外周血弓形虫循环抗原CAg的表达变化;采用双基因PCR法检测弓形虫RH株速殖子在小鼠腹腔和脑、心、肺、肝、脾、肾等脏器中的增殖变化,并制备病理切片,染色后观察各脏器组织病理学变化。结果复方一枝蒿颗粒能够适当延长小鼠生存时间,延长率17.74%。组织学变化显示药物可减轻小鼠肺脏组织瘀血,并使小鼠脾脏出现大量多核型巨细胞形态。复方一枝蒿颗粒可抑制虫体在腹腔和肝、肾、心、肺组织中的增殖,感染4d时对小鼠腹腔冲洗液中游离速殖子的增殖抑制率可达92.80%,联用阿奇霉素增值抑制率达98.53%。相对定量PCR法显示弓形虫复方一枝蒿颗粒对小鼠肝、肾、心、肺组织中弓形虫增殖抑制率分别为(79.60±0.08)%、(42.00±0.20)%、(50.80±0.08)%、(60.40±0.21)%,作用虽弱于阿奇霉素对照组和联合药物组,但可显著抑制小鼠脑组织中弓形虫的增殖,增值抑制率达(76.80±0.07)%(F=63.08,P<0.01);核酸检测弓形虫529基因比B1基因在小鼠脏器中的表达更稳定,适宜作为弓形虫在宿主脏器组织中的监测指标。复方一枝蒿颗粒能降低虫体循环抗原CAg在感染小鼠外周血清中的表达水平,与感染组小鼠相比(3 970.82±20.52)pg/ml,至感染4d时药物组小鼠血清中CAg水平为(3 163.01±15.94)pg/ml。结论复方一枝蒿颗粒单用或联合阿奇霉素使用对小鼠急性弓形虫感染有一定治疗效果。与阿奇霉素对虫体的直接杀伤作用不同,复方一枝蒿颗粒主要通过增强脾脏免疫细胞吞噬作用和抑制虫体向脑能远端脏器的侵袭抑制感染鼠的病情发展,推测可能通过调节小鼠免疫功能起到治疗小鼠急性弓形虫感染的作用。
Objective To explore the effect of Yizhihao granules containing Artemisia rupestris L.alone or in combination with azithromycin on acute Toxoplasma gondii infection in mice. Methods An animal model of acute T.gondii infection in KM mice was created by intraperitoneally injecting tachyzoites from the RH strain of T.gondii,and mice were randomly divided into 4 groups:an acute T.gondii infection control group,a Yizhihao granules group,a Yizhihao granules+azithromycin group,and an azithromycin control group.Symptoms in and the survival of mice were observed.Expression of T.gondii circulating antigen(Cag)in the peripheral blood of mice was detected using ELISA.Expression of the Tg529 gene and the B1 gene,found in the abdominal cavity and organs such as the brain,heart,spleen,kidneys,lungs,and liver of mice,was detected using dual-gene PCR.Histopathological changes in each organ were observed after pathological sections were stained. Results Yizhihao granules adequately prolonged the survival of mice 17.74%.Histological changes indicated that the drug reduced blood stasis in the lung tissue of mice and produced a morphology of numerous multinucleated giant cells in the spleens of mice.Yizhihao granules inhibited the proliferation of T.gondii in the abdominal cavity and liver,kidney,heart,and lung tissues.Four d after infection,inhibition of free tachyzoite proliferation in the peritoneal lavage fluid of mice reached 92.80%,and inhibition of proliferation in the Yizhihao granules+azithromycin group reached 98.53%.Although the effect was weaker in the azithromycin control group and the combined drug group,relative quantitative PCR indicated that Yizhihao granules inhibited T.gondii proliferation in the liver,kidney,heart,and lung tissues of mice.Inhibition of Tg529 expression was 79.60±0.08%in the liver,42.00±0.20%in the kidneys,50.80±0.08%in the heart,and 60.40±0.21%in the lungs.The drug significantly inhibited the proliferation of T.gondiiin the brain tissues of mice at a rate of 76.80±0.07%(F=63.08,P<0.01).Expression of the Tg529 gene was more stable than that of the B1 gene after detection using PCR,which is suitable for monitoring indicators of T.gondii infection in organ tissues of a host.Yizhihao granules reduced the level of CAg expression in the serum of infected mice.Compared to the infected group(3 970.82±20.52)pg/m,the CAg level in the serum of the treated group was 3,163.01±15.94 pg/ml 4 days after infection. Conclusion Yizhihao granules alone or in combination with azithromycin had a certain therapeutic effect on acute Toxoplasmainfection in mice.Unlike the direct killing of a parasite by azithromycin,Yizhihao granules mainly enhance phagocytosis by spleen immune cells and inhibit T.gondii from invading distal organs such as the brain.Presumably,the drug may treat disease by regulating immune function in mice.
作者
王宇靖
郭琼
唐云剑
李莹花
高剑
齐瑞瑞
巴克骄
WANG Yu-jing;GUO Qiong;TANG Yun-jian;LI Ying-hua;GAO Jian;QI Rui-rui;BA Kejiao(Class graduating in 2015-5,Clinical Medicine,First College of Clinical Medicine,Xinjiang Medical University,Urumqi,China 830011;College of Basic Medicine,Xinjiang Medical University)
出处
《中国病原生物学杂志》
CSCD
北大核心
2020年第2期195-201,共7页
Journal of Pathogen Biology
基金
新疆维吾尔自治区科技厅科技计划联合基金项目(No.2016D01C186)
新疆维吾尔自治区级大学生创新性实验训练计划(No.CX201710760040)。
关键词
复方一枝蒿颗粒
急性弓形虫感染
增殖抑制
双基因PCR
弓形虫529基因
Yizhihao granules containing Artemisia rupestris L.
acute Toxoplasma infection
inhibition of proliferation
dual—gene PCR
Toxoplasma 529 gene
