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Outline
To observe the effect of Anchang Decoction (安肠汤, ACD) on mitophagy in intestinal epithelial cells of the rats with ulcerative colitis (UC) through PTEN inducing putative kinase 1 (PINK1) /Parkinson's disease protein (Parkin) /antiproliferative protein 2 (PHB2) /phosphoglycinate mutase 5 (PGAM5) signaling axis.
Forty-eight Wistar rats were randomly divided into Control group, Model group, Mesalazine group (0.42 g·kg-1) and ACD low, medium and high groups (ACD-L, ACD-M, ACD-H; 11.55, 23.1, 46.2 g·kg-1), 8 rats in each group. Except for the Control group, the UC model was established by 2, 4, 6-trinitrobenzenesulfonic acid (TNBS) . After the model was successfully established, the blank group and the model group were given the normal saline by gavage, and the other groups were given the corresponding drugs by gavage for 14 days. After the intervention, the general condition of the rats in each group was observed and the body weight was recorded, the disease activity index (DAI) score was calculated, the pathological changes of intestinal tract were observed by hematoxylin-eosin staining, and the levels of interleukin-6 (IL-6) and interleukin-18 (IL-18) were detected by enzyme-linked immunosorbent assay. The mitochondrial ultrastructure of colonic epithelial cells (IECs) was observed by transmission electron microscopy. The expressions of mitochondrial microtubule-associated protein 1 light chain 3 (LC3) and selective autophagy receptor (p62) were detected by immunofluorescence. The protein expression levels of PHB2, PGAM5, PINK1, Parkin, LC3 and p62 in colon tissues were detected by Western Blot. The mRNA expression levels of PHB2, PGAM5, PINK1 and Parkin in colon tissues were detected by Real-time fluorescent quantitative polymerase chain reaction.
Compared with those of the Control group, the general condition of the Model group was worse, the body weight was significantly decreased, the DAI score was significantly increased (P<0.01), the pathological changes of colon tissue were obvious, the serum levels of IL-6 and IL-18 were increased (P<0.01), the mitochondrial structure of IECs was obviously missing, the number of Iecs was decreased, and the mitochondrial cristae was broken. The mitochondrial morphology was swollen and vacuolated, the protein and mRNA expression levels of LC3, PHB2, PGAM5, PINK1 and Parkin were decreased, and the protein expression level of p62 was increased in the treatment group (P<0.05, P<0.01) . Compared with those in the Model group, the rats in the Mesalazine group and each dose group of ACD were significantly improved, the body weight increased, the DAI score increased significantly (P<0.05, P<0.01), the colonic mucosal structure recovered to varying degrees, and the serum levels of IL-6 and IL-18 decreased (P<0.05, P<0.01) . The mitochondrial membrane of IECs was more complete, the number was increased, the swelling degree was decreased, and the structure of mitochondrial cristae was improved. ACD-Mand ACD-H showed more obvious autophagosomes or autolysosomes. The expression levels of LC3 protein, PHB2, PGAM5, PINK1 and Parkin protein and mRNA were increased in varying degrees. The p62 protein levels were decreased in different degrees (P<0.05, P<0.01) .
ACD reduces inflammatory response and promotes intestinal mucosal recovery to improve UC symptoms possibly by regulating the PHB2/PGAM5/PINK1/Parkin axis to promote mitophagy in IECs.
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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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