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To identify the flavonoids in persimmon leaf (PL) extract, and to explore the mechanism of the action on ulcerative colitis (UC) from the perspective of TLR4/NLRP3 inflammatory signal pathway and intestinal flora.
HPLC was used to determine the five flavonoids in persimmon leaf extract. The animal model of UC mice was established by 3% dextran sulfate sodium (DSS). The body weight and drinking volume of the mice were measured daily. The detection of IL-6, TNF-α, IL-1β and IL-10 in the serum of the mice were measured by Elisa kit, the changes of colon tissue were observed by HE staining, the expression levels of TLR4, IL-1β, NF-κB p65 and NLRP3 proteins in the colon tissues were determined by western blot, and the changes of intestinal flora in feces were detected by 16S rDNA amplification sequencing.
The body weight, colon length and IL-10 level in DSS group were significantly lower than that in control group (P<0.01). The disease activity index and the levels of IL-6, TNF-α and IL-1β in DSS group were significantly increased (P<0.01) compared with the control group. Compared with the DSS group, the body weight decreased slowly, the colon length prolonged and the disease activity index decreased in the treated group, the levels of IL-6, TNF-α and IL-1β significantly reduced (P<0.01) and IL-10 significantly increased (P<0.01). The expression levels of TLR4/NLRP3-related proteins significantly increased (P<0.01), the expression levels of TLR4, IL-1β, NF-κB p65 and NLRP3 proteins significantly decreased (P<0.01). The results of 16S rDNA amplification sequencing showed that Lactobacillus murinus, Lactobacillus johnsonii, Romboutsia and Akkermansia muciniphila had an increasing trend in the treated group. The pathological results showed that there were severe mucosal ulcer, crypt loss, inflammatory cell infiltration and goblet cell depletion in colon tissue of mice in DSS group and the pathological changes of colon tissue in mice were improved after administration. The HPLC method has good separation degree and high accuracy.
Persimmon leaf extract has protective effect on intestinal injury in DSS-induced UC mice. It may improve the intestinal microenvironment by inhibiting TLR4/NLRP3 inflammatory signal pathway and relieving inflammation.
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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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