Wang Xinjian, Yang Zhen, Zhang Yujun, Cheng Feng, Xing Xiaoyong, Wen Fengqin, Hu Yonghao, Chen Changjiang, Wei Bin, Bai Pengxia, Wang Xuehong, Liu Yu, Zhang Hongjuan, Hao Baocheng, Wang Shengyi
Key Laboratory of New Animal Drug Project, Gansu Province, Key Laboratory of Veterinary Pharmaceutical Development, Ministry of Agriculture and Rural Affairs, Lanzhou Institute of Husbandry and Pharmaceutical Sciences of Chinese Academy of Agriculture Sciences, Lanzhou, China.
College of Veterinary Medicine, Gansu Agricultural University, Lanzhou, China.
Front Vet Sci. 2022 Jul 27;9:951058. doi: 10.3389/fvets.2022.951058. eCollection 2022.
disease is a common ectoparasitosis, which can lead to a decrease in animal production performance, product quality, and even death. Hand.-Mazz. has many pharmacological activities including insecticidal, heat-clearing, analgesic, and dehumidifying. However, there are few researches focused on the effects and related mechanism of Hand.-Mazz. in killing . In this study, 11 alkaloids of Hand.-Mazz. were detected, and its total alkaloid activity was determined. The results showed when the total alkaloid concentration was 64 mg/ml and the treatment time was 16 h, the killing rate of reached 100%. Through the observation of the differences in the surface of in each experimental group, it was found that the morphology of the posterior end of the female in the alkaloid treatment group was significantly different from that of the blank and positive control groups, and most of the epidermal tissue was obsessive and missing. Moreover, the enzyme activity determination results of 64 mg/ml group were significantly different when compared with the normal control group, while there was no significant difference in other groups. Then, the gene library was established by the unreferenced genome transcriptome sequencing, the proteomic comparison was performed using tandem mass tag labeled protein detection technology, and finally, the samples were quantitatively analyzed by liquid chromatography-mass spectrometry tandem and bioinformatics methods. Based on the above experimental results, it was speculated that Hand.-Mazz. total alkaloids may cause the imbalance of protein disulfide isomerase expressions by affecting the regulation of Hsp40 cellular protein homeostasis and the oxidation of protein disulfide isomerase and related proteins. This would affect the selective recognition of signal sequence, the targeted transport of Sec 61, and the correct folding of the three-dimensional structure of amino acid chain, weakening the clearance of amino acid chains that cannot be correctly folded and eventually resulting in the killing of . This study preliminarily revealed the mechanism of Hand.-Mazz. total alkaloids against and provided a theoretical basis for the screening of action targets and the development of new veterinary drugs.
疾病是一种常见的体外寄生虫病，可导致动物生产性能下降、产品质量降低，甚至死亡。[植物名称，因未给出中文正式名称，保留原文Hand.-Mazz.]具有多种药理活性，包括杀虫、清热、止痛和除湿。然而，针对[植物名称，因未给出中文正式名称，保留原文Hand.-Mazz.]杀虫作用及其相关机制的研究较少。本研究检测了[植物名称，因未给出中文正式名称，保留原文Hand.-Mazz.]中的11种生物碱，并测定了其总生物碱活性。结果表明，当总生物碱浓度为64mg/ml且处理时间为16h时，[寄生虫名称，因未给出中文正式名称，保留原文]的杀灭率达到100%。通过观察各实验组[寄生虫名称，因未给出中文正式名称，保留原文]体表差异，发现生物碱处理组雌虫后端形态与空白对照组和阳性对照组有显著差异，大部分表皮组织脱落缺失。此外，64mg/ml组的酶活性测定结果与正常对照组相比有显著差异，而其他组无显著差异。然后，通过无参考基因组转录组测序建立[寄生虫名称，因未给出中文正式名称，保留原文]基因文库，采用串联质谱标签标记蛋白质检测技术进行蛋白质组比较，最后通过液相色谱-质谱联用和生物信息学方法对样本进行定量分析。基于上述实验结果推测，[植物名称，因未给出中文正式名称，保留原文Hand.-Mazz.]总生物碱可能通过影响Hsp40细胞蛋白质稳态调控和蛋白质二硫键异构酶及相关蛋白的氧化，导致蛋白质二硫键异构酶表达失衡，进而影响信号序列的选择性识别、Sec 61的靶向转运以及氨基酸链三维结构的正确折叠，削弱对不能正确折叠的氨基酸链的清除能力，最终导致[寄生虫名称，因未给出中文正式名称，保留原文]死亡。本研究初步揭示了[植物名称，因未给出中文正式名称，保留原文Hand.-Mazz.]总生物碱对[寄生虫名称，因未给出中文正式名称，保留原文]的作用机制，为筛选[寄生虫名称，因未给出中文正式名称，保留原文]作用靶点和开发新型兽药提供了理论依据。
