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To examine the impact of drinking water arsenic exposure on the development of generalized anxiety disorder (GAD) and to investigate specific changes in plasma exosomal miRNAs.
Sixteen male SPF-grade Wistar rats were randomly divided into two groups: control and arsenic. Each group consisted of 8 rats. The arsenic group was exposed to 0.10 mg/L. All groups had unrestricted access to drinking water. The rats were exposed to arsenic for a duration of 21 days. Behavioral assessments using the open field and elevated plus maze tests were conducted on days 0, 7, 14, and 21 to evaluate GAD-like behavior. At the end of the experiment, plasma exosomal miRNAs were extracted from both groups of rats. Sequencing was performed, and the expression levels of differentially expressed miRNAs were validated using real-time quantitative PCR.
On the 21st day after arsenic exposure, the arsenic group exhibited lower body weight compared to the control group (t=7.950, P<0.001). In the open field test, the arsenic group showed a decrease in total distance traveled (t=2.213, P=0.044), an increase in rearing frequency (t=-4.704, P<0.001), an increase in grid crossings (t=4.340, P=0.001), and a decrease in standing frequency (t=4.496, P=0.001) compared to the control group. In the elevated plus maze test, the arsenic group exhibited a decrease in the number of entries into the open arms (t=3.614, P=0.003) and a decrease in time spent in the open arms (t=4.775, P<0.001), while showing an increase in the number of entries into the closed arms (t=-2.486, P=0.026) and an increase in time spent in the closed arms (t=-6.862, P<0.001) compared to the control group. These results indicate GAD-like behavior in the rats exposed to arsenic. Real-time quantitative PCR revealed that the expression levels of miR-99b-3p, miR-9a-5p, and miR-218a-5p were upregulated, while the expression levels of miR-425a-3p, miR-378a-3p, and miR-155-5p were downregulated in the arsenic group compared to the control group (P<0.05). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of the differentially expressed plasma exosomal miRNAs indicated their potential involvement in GAD development through processes such as endocytosis, MAPK pathway, Ras signaling pathway, cAMP signaling pathway, mTOR signaling pathway, and apoptosis.
Drinking water arsenic exposure can induce GAD-like behavior in rats. Plasma exosomal miRNAs, including miR-99b-3p, miR-9a-5p, miR-218a-5p, miR-425a-3p, miR-378a-3p, and miR-155-5p, may be involved in the regulation of GAD development induced by drinking water arsenic exposure.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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