体外杀线虫作用及青蒿素对沙土鼠捻转血矛线虫的驱虫活性和过渡幼虫（L-L）中 Hc29 基因的相对表达
In Vitro Nematocidal Effect and Anthelmintic Activity of Artemisia cina Against Haemonchus contortus in Gerbils and Relative Expression of Hc29 Gene in Transitional Larvae (L-L).
机构信息
Facultad de Estudios Superiores Cuautitlán, Universidad Nacional Autónoma de México, Mexico, Estado de México, Mexico.
Centro Nacional de Investigación Disciplinaria en Salud Animal e Inocuidad, Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias (CENID-SAI, INIFAP), Carr. Fed. Cuernavaca-Cuautla 8534, CP 62550, Jiutepec, Morelos, Mexico.
出版信息
Acta Parasitol. 2021 Sep;66(3):938-946. doi: 10.1007/s11686-021-00364-w. Epub 2021 Mar 15.
PURPOSE
(1) To assess the in vitro activity of Artemisia cina against Haemonchus contortus L (HcL) and in transitional (L-L) larvae (HcTrL-L); (2) to quantify the relative expression of the Hc29 gene in HcTrL-L exposed to the A. cina n-hexane extract; and (3) to assess the anthelmintic activity (AA) of the A. cina organic extracts in gerbils artificially infected with H. contortus (HcArt/inf/gerbs).
METHODS
The in vitro assay was carried out in 96-well microtitration plates. The following A. cina extracts: ethyl acetate (Ac-EtOAcEx), n-hexane (Ac-n-HexEx), and methanol (Ac-MethEx) were assessed at 1 and 2 mg/mL against HcL and HcTrL-L at 24 h exposure. Relative expression of the Hc29 gene in HcTrL-L was obtained by RT-PCR. For assessing the AA, six groups of five HcArt/inf/gerbs were used. Groups were treated orally with 4 mg/kg BW of A. cina extracts. Five days after treatment, the gerbils were necropsied and nematodes counted.
RESULTS
The highest in vitro activities (75 and 82.6%) were shown by Ac-n-HexEx at 1 and 2 mg/mL, respectively. For HcTrL-L the highest in vitro activities (69 and 23%) were shown by Ac-n-HexEx and isoguaiacine at 0.625 mg/mL, respectively. Also, upregulation of H. contortus Hc29 gene by 13- and 80-fold (p < 0.01) was observed on the HcTrL-L stage after exposure to Ac-n-HexEx extract and isoguaiacine at 0.078 mg/mL, respectively. Reduction percentage was 100% in HcArt/inf/gerbs treated with Ac-n-HexEx.
CONCLUSIONS
We conclude that the Ac-n-HexEx and isoguaiacine compound had anthelmintic efficacy against H. contortus and L and HcTrL-L.
目的
（1）评估青蒿素对捻转血矛线虫（HcL）和过渡性（L-L）幼虫（HcTrL-L）的体外活性；（2）定量暴露于青蒿素正己烷提取物的 HcTrL-L 中 Hc29 基因的相对表达；（3）评估青蒿素有机提取物在人工感染捻转血矛线虫的沙鼠中的驱虫活性（AA）（HcArt/inf/gerbs）。
方法
在 96 孔微量滴定板中进行体外测定。评估以下青蒿素提取物：乙酸乙酯（Ac-EtOAcEx）、正己烷（Ac-n-HexEx）和甲醇（Ac-MethEx）在 24 小时暴露时对 HcL 和 HcTrL-L 的浓度为 1 和 2mg/mL。通过 RT-PCR 获得 HcTrL-L 中 Hc29 基因的相对表达。为了评估 AA，使用了六组五只 HcArt/inf/gerbs。组用 4mg/kg BW 的青蒿素提取物口服治疗。治疗后 5 天，对沙鼠进行尸检并计数线虫。
结果
Ac-n-HexEx 在 1 和 2mg/mL 时显示出最高的体外活性（75%和 82.6%）。对于 HcTrL-L，Ac-n-HexEx 和异愈创木酚在 0.625mg/mL 时显示出最高的体外活性（69%和 23%）。此外，在暴露于正己烷提取物和异愈创木酚（0.078mg/mL）后，HcTrL-L 阶段的 H. contortus Hc29 基因的上调分别为 13 倍和 80 倍（p<0.01）。用 Ac-n-HexEx 处理的 HcArt/inf/gerbs 的减少百分比为 100%。
结论
我们得出结论，Ac-n-HexEx 和异愈创木酚化合物对捻转血矛线虫和 L 和 HcTrL-L 具有驱虫作用。
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