Objective: To compare the differences in pathogenicity and gene expression profiles between adult Schistosoma japonicum isolated from hilly and marshland and lake regions of Anhui Province, so as to provide the scientific evidence for formulating the precise schistosomiasis control strategy in different endemic foci.
Methods: C57BL/6 mice were infected with cercariae of S. japonicum isolates from Shitai County (hilly regions) and Susong County (marshland and lake regions) of Anhui Province in 2021, and all mice were sacrificed 44 days post-infection and dissected. The worm burdens, number of S. japonicum eggs deposited in the liver, and the area of egg granulomas in the liver were measured to compare the difference in the pathogenicity between the two isolates. In addition, female and male adult S. japonicum worms were collected and subjected to transcriptome sequencing, and the gene expression profiles were compared between Shitai and Susong isolates of S. japonicum. The differentially expressed genes (DEGs) were subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses.
Results: The total worm burdens [(14.50 ± 3.96) worms/mouse vs. (16.10 ± 3.78) worms/mouse; t = 0.877, P = 0.392], number of female and male paired worms [(4.50 ± 0.67) worms/mouse vs. (5.10 ± 1.45) worms/mouse; t = 1.129, P = 0.280], number of unpaired male worms [(5.50 ± 4.01) worms/mouse vs. (5.60 ± 1.69) worms/mouse; t = 0.069, P = 0.946], number of eggs deposited in per gram liver [(12 116.70 ± 6 508.83) eggs vs. (16 696.70 ± 4 571.56) eggs; t = 1.821, P = 0.085], and area of a single egg granuloma in the liver [(74 359.40 ± 11 766.34) µm2 vs. (74 836.90 ± 13 086.12) µm2; t = 0.081, P = 0.936] were comparable between Shitai and Susong isolates of S. japonicum. Transcriptome sequencing identified 584 DEGs between adult female worms and 1 598 DEGs between adult male worms of Shitai and Susong isolates of S. japonicum. GO enrichment analysis showed that the DEGs between female adults were predominantly enriched in biological processes of stimulus response, cytotoxicity, multiple cell biological processes, metabolic processes, cellular processes and signaling pathways, cellular components of cell, organelles and cell membranes and molecular functions of binding and catalytic ability, and KEGG enrichment analysis showed that these DEGs were significantly enriched in pathways of vascular endothelial growth factor signaling, glutathione metabolism, arginine and proline metabolism. In addition, the DEGs between male adults were predominantly enriched in biological processes of signaling transduction, multiple cell biological processes, regulation of biological processes, metabolic processes, development processes and stimulus responses, cellular components of extracellular matrix and cell junction and molecular functions of binding and catalytic ability, and these DEGs were significantly enriched in pathways of Wnt signaling, Ras signaling, natural killer cells-mediated cytotoxicity, extracellular matrix-receptor interactions and arginine biosynthesis.
Conclusions: There is no significant difference in the pathogenicity between S. japonicum isolates from hilly and marshland and lake regions of Anhui Province; however, the gene expression profiles vary significantly between S. japonicum isolates.
［摘要］ 目的比较安徽省山丘地区和湖沼地区日本血吸虫致病性及成虫基因表达谱差异, 为制定不同流行区血吸虫 病精准防治策略提供科学依据。方法 2021年分别采用来源于安徽省山丘地区石台县和湖沼地区宿松县的日本血吸 虫尾呦感染C57BL/6小鼠, 44 d后处死并解剖小鼠, 比较两组小鼠虫荷数、肝脏沉积虫卵数和肝脏虫卵肉芽肿面积等指 标, 观察山丘地区和湖沼地区日本血吸虫虫株对小鼠致病性差异。对两组小鼠体内分离的雌性和雄性日本血吸虫成虫 进行转录组测序, 比较山丘地区和湖沼地区日本血吸虫虫株基因表达谱差异, 对差异表达基因进行基因本体论(GO)和 京都基因和基因组百科全书(IŒGG)富集分析。结果 安徽省山丘地区和湖沼地区日本血吸虫尾呦感染小鼠44 d后, 小 鼠体内总虫荷数分别为(14.50 ± 3.96)条/鼠和(16.10 ± 3.78)条/鼠(t = 0.877, P = 0.392)、雌雄合抱对数分别为(4.50 ± 0.67)对/鼠和(5.10 ± 1.45)对/鼠(t = 1.129, P = 0.280)、未合抱雄虫数分别为(5.50 ± 4.01)条/鼠和(5.60 ± 1.69)条/鼠(t = 0.069, P = 0.946)、每克肝脏沉积虫卵数分别为(12 116.70 ± 6 508.83)只和(16 696.70 ± 4 571.56)只(t = 1.821, P = 0.085)』干脏单个虫卵肉芽肿面积分别为(74 359.40 ± 11 766.34) µm2和(74 836.90 ± 13 086.12) µm2 (t = 0.081, P = 0.936), 差异均无统计学意义。转录组测序表明, 安徽省山丘地区和湖沼地区日本血吸虫雌性和雄性成虫间分别存在 584个和1 598个差异表达基因。GO富集分析显示, 雌虫间差异表达基因主要参与刺激反应、细胞杀伤、多细胞生物进 程、代谢进程、细胞进程及信号通路等生物学过程, 细胞、细胞器和细胞膜等细胞组分, 结合和催化活性等分子功能; KEGG通路分析显示, 差异表达基因显著富集于血管内皮细胞生长因子(VEGF)信号通路、谷胱甘肽代谢、精氨酸和脯氨 酸代谢等信号通路。GO富集分析显示, 雄虫间差异表达基因主要参与信号传导、多细胞生物进程、生物过程调节、代谢 过程、发育过程、刺激反应等生物学过程, 细胞外基质、细胞连接等细胞组分, 结合和催化活性等分子功能;KEGG通路分 析显示, 差异表达基因显著富集于Wnt信号通路、Ras信号通路、自然杀伤细胞介导的细胞毒性、细胞外基质-受体相互作 用、精氨酸生物合成等信号通路。结论 安徽省山丘地区和湖沼地区日本血吸虫致病性差异无统计学意义, 但虫株基因 表达谱存在显著差异。.
Keywords:
Gene expression profile; Hilly region; Marshland and lake region; Pathogenicity;