To investigate the relationship between the inflammatory state of obesity with phlegm-dampness syndrome and the M1 polarization of islet macrophages, as well as the intervention mechanism of Wendan Decoction.
Forty-five SD rats were randomly divided into a blank group and a modeling group. The modeling group was fed a high-fat diet for 8 weeks to establish an obesity with phlegm-dampness syndrome model. Twenty obese rats were then selected and divided into a model group, a Xuezhikang group, and low-/high-dose Wendan Decoction groups (5 rats each). The blank group (normal group) comprised 5 rats. Each treatment group received corresponding doses via gavage for 6 weeks, while the model and normal groups received equal volumes of distilled water. Body weight, Lee’s index, and lipid levels were measured. ELISA was used to determine pro-inflammatory cytokines (TNF-α, IL-6, IFN-γ) and fasting insulin (FINS) in islet tissue. HE staining and immunohistochemistry were employed to observe islet histopathology and the expression of macrophage marker CD68 (Cluster of Differentiation 68), M1 activation marker CD86 (Cluster of Differentiation 86), and iNOS (inducible nitric oxide synthase) . Western blot was used to detect CD68, CD86, and iNOS protein expression.
After successful modeling, pathological improvements were observed in all treatment groups. Compared with the normal group, the model group showed significant changed in body weight, Lee’s index, lipid levels (elevated TG and LDL-C, decreased HDL-C) (P<0.001 or P<0.01), FINS, HOMA-IR, and TNF-α/IL-6 secretion (P<0.05、P<0.01 or P<0.001) . Compared with the model group, all treatment groups exhibited significant reductions in body weight, obesity rate, TG, FINS, HOMA-IR, and TNF-α/IL-6 secretion (P<0.01、P<0.05 or P<0.001). The high-dose Wendan Decoction group showed significant decreases in Lee’s index and LDL-C (P<0.01 or P<0.05) . Histopathological and immunohistochemical analyses showed that the infiltration of CD68, CD86, and iNOS positive cells in the islet interstitium was significantly increased in the model group (P<0.05) ; the integrity of islet tissue was improved in each treatment group, with significantly reduced infiltration of CD68, CD86, and iNOS positive cells (P<0.05 or P<0.01) . Among them, only a small number of positive cells were occasionally observed in the low-/high-dose Wendan Decoction groups, and the number of positive cells was relatively fewer in the Xuezhikang group. Western blot (WB) assay revealed that compared with the normal group, the protein expression levels of CD68, CD86, and iNOS in the islets were significantly elevated in the model group (P<0.000 1) ; compared with the model group, the expression of CD86 and iNOS proteins was significantly reduced in the low-dose Wendan Decoction group (P<0.05 or P<0.001), while the expression of CD68, CD86, and iNOS proteins was significantly decreased in both the high-dose Wendan Decoction group and the Xuezhikang group (P<0.001). The high-dose Wendan Decoction group outperformed the Xuezhikang group in lowering Lee’s index and CD68 protein expression (P<0.05 or P<0.001) .
The inflammatory state of obesity with phlegm-dampness syndrome is closely associated with M1 polarization of islet macrophages (manifested as abnormal expression of CD68+, CD86+, and iNOS+) . Wendan Decoction can significantly improve the inflammatory state of obesity with phlegm-dampness syndrome by regulating M1 polarization of islet macrophages.
| 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 |