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Objective To investigate the role and significance of abdominal paracentesis drainage (APD) to pyroptosis in the pancreas of rats with severe acute pancreatitis (SAP), which is mediated by NLRP3 inflammasome-activated-caspase-1. Methods A total of 48 healthy male SD rats were randomly divided into three groups: sham operation (Sham) group, SAP group, and APD group, with 16 rats in each group. 5% sodium sulfonate was retrogradely injected into the pancreaticobiliary duct to establish the SAP model. In the APD group, besides the SAP induction, a drainage tube was placed on the right lower abdomen for drainage. Serum and pancreatic tissue were collected 12 hours after modeling. We examined the activities of serum lipase and amylase using an automatic biochemical analyzer and evaluated pancreatic damage through HE staining. In addition, we quantified the expression of serum inflammatory factors by ELISA and measured the expression level of genes and proteins related to caspase-1 mediated pyroptosis in pancreatic tissue using RT-PCR and Western blotting. Lastly, we observed the structural changes of the subcellular organelles and the characteristic changes of pyrolysis in pancreatic acinar cells using transmission electron microscopy. Results Compared with the SAP group, the pancreas tissue in the ADP group showed alleviated damages with a much lower pathological score; the levels of serum lipase, amylase, tumor necrosis factor(TNF)-α, Interleukin(IL)-6, IL-1β, and IL-18 also showed significant reduction (P<0.05).Compared with the Sham group, the expression levels of NLRP3, ASC, Caspase-1, cleaved caspase-1, GSDMD, and cleaved-GSDMD were all significantly up-regulated in the pancreas in the SAP group. After APD treatment, the expression of these Caspase-1-mediated pyrolysis pathway key genes in pancreatic tissue was significantly down-regulated. In addition, the expression levels of IL-1β and IL-18 mRNA in pancreatic tissue were also significantly reduced (P<0.05). Transmission electron microscopy showed that the SAP group had endoplasmic reticulum and mitochondrial expansion, chromatin condensation, and the characteristic change of pyrolysis, namely the formation of membrane pores in the cell membrane. In the APD group, we observed reduced chromatin condensation in the nucleus and less expansion in the endoplasmic reticulum and mitochondrial. Conclusion SAP showed activated Caspase-1-mediated pyroptosis. Early treatment with APD can alleviate the severity of SAP, possibly by inhibiting the activation of Caspase-1-mediated pyroptosis, thereby reducing local and systemic inflammatory reactions.
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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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