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To construct a Propionibacterium acnes (P. acnes)-induced cellular inflammation model and to study the anti-inflammatory effect of a thermal water.
Utilize human immortalized keratinocytes (HaCaT) was used in the experimentation. Co-incubate the cells with thermal spring water and employ the CCK-8 assay to assess cell viability, screening for the optimal intervention time for the thermal spring water. Expose the cells to varying concentrations of P. acnes for different times, measure cell viability, calculate the 50% inhibitory concentration (IC50) of P. acnes, and determine the model induction conditions by testing IL-6 levels. The cells were soaked in thermal spring water for 0.5 hours, followed by co-culturing with P. acnes as the preventive intervention group, or co-culturing with P. acnes first and then soaking the cells in hot spring water for 0.5 hours as the therapeutic intervention group. The levels of cell viability, pro-inflammatory cytokines, and gene expression of inflammation pathways were assessed using the CCK-8 assay, ELISA, and RT-PCR, respectively. Concurrently, control groups with saline solution, tap water, and pure water, along with model and blank controlswere set up.
After treating the cells with thermal spring water and physiological saline for 0.5 hours, cell viabilities were 92.7% and 99.09%, respectively, which led to the selection of 0.5 hours as the intervention time for subsequent experiments.P. acnes at an OD600 of 0.25 was co-incubated with the cells for 5 hours to establish the model. In the preventive intervention, the secretion levels of IL-1β and IL-8， as well as the relative expression levels of IL-6, TNF-α, IL-8, and the genes TLR2 and NF-κB in the cells of the thermal spring water group, were all lower than those in the model group (P<0.05), and significantly lower than those in the saline solution group. In the therapeutic intervention, the contents of IL-6, IL-1β, TNF-α, and IL-8, as well as the expression levels of the genes IL-6, IL-1β, TNF-α, TLR2, and NF-κB in the thermal spring water group, were all lower than those in the model group. The contents of IL-6, IL-1β, TNF-α, and the expression levels of the genes IL-6, IL-1β, IL-8, TLR2, and NF-κB were also significantly reduced compared to those in the saline solution group (P<0.05).
This thermal water may exert its anti-inflammatory effect by down-regulating the expression of TLR2 and NF-κB genes in inflammatory signalling pathway and by reducing the secretion of pro-inflammatory cytokines.
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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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