| OBJECTIVE: To study the changes of intestinal microfold cells(M cells)in the early stage of severe acute pancreatitis(SAP)in mice,explore the effect of Dai-Huang-Fu-Zi-Tang intervention on M cells,and reveal the mechanism of protecting the intestinal immune barrier.Preventing SAP lays a theoretical foundation.Methods: Forty-eight healthy SPF C57BL/6J male mice weighing 30-35 g and 8 weeks of age were randomly divided into 4 groups: blank control group(10 mice),rhubarb aconite soup control group(10 mice),and SAP group(10),Dai-Huang-Fu-Zi-Tang treatment group(10),isoflurane gas anesthesia,the experimental group was intraperitoneally injected with 20% L-arginine at a dose of 3.5g/kg,the control group was intraperitoneally injected with the same amount of normal saline,the blank control group and SAP The group was given saline 0.2ml enema at 12,24 and 36 h after modeling,and the control group and the treatment group of Dai-Huang-Fu-Zi-Tang were given 0.2ml enema at 12,24 and 36 h after modeling.After 48 hours of modeling,serum was taken and the content of serum amylase,endotoxin,IL-1β and TNF-α was detected by ELISA method;the ileum,pancreas and lung tissue were taken for HE staining,and the pancreas tissue was scored by the modified Schmidt scoring standard for pathological scoring.The ileal tissues were scored by Chiu intestinal mucosal injury scoring criteria for pathology,and the lung tissues were scored by Smith intestinal mucosal injury scoring criteria for pathology score;the ileum was also taken for immunohistochemical staining to detect the changes of M cell-specific protein GP2.Results: 1.Among the four groups of mice,all the two control groups survived,three died in the SAP group,and one died in the treatment group of Dai-Huang-Fu-Zi-Tang.The mice in the control group generally performed well after intraperitoneal injection,with normal drinking water and free movement.The mice in the experimental group generallyhad poor conditions after intraperitoneal injection,lack of energy,decreased drinking water,unresponsiveness,slow movement,shortness of breath,and dorsal abdomen.2.There was no significant difference in the levels of amylase,endotoxin,IL-1β and TNF-α between the two control groups;compared with the blank control group,the levels of amylase,endotoxin,IL-1β and TNF-α in the SAP group increased significantly High;Compared with SAP group,the treatment group of Dai-Huang-Fu-Zi-Tang significantly decreased the levels of serum amylase,endotoxin,IL-1β and TNF-α.3.General observation and HE staining: macroscopically,the pancreas,ileum,and lungs of the control group were normal;the SAP group showed pancreas,ileum,and lung edema and congestion,and local necrosis;and the pancreas,ileum,and lungs of the treatment group of Dai-Huang-Fu-Zi-Tang were slightly injured.Microscopically,the pancreas,ileum,and lung tissue in the control group were normal;the pancreas,ileum,and lung tissue in the SAP group were congested and necrotic;the pancreatic,ileal,and lung tissues in the treatment group of Dai-Huang-Fu-Zi-Tang were mildly congested and necrotic.Compared with the blank control group,the histopathology scores of the pancreas,ileum,and lung were significantly increased in the SAP group(P < 0.05);the histopathology scores of the pancreas,ileum,and lung were significantly reduced in the treatment group of Dai-Huang-Fu-Zi-Tang compared with the SAP group(P<0.05).4.Immunohistochemical staining of M cell-specific protein GP2: Compared with the blank control group,the Dai-Huang-Fu-Zi-Tang control group had no significant difference in GP2 levels;compared with the blank control group,the GP group expression of the SAP group was significantly reduced(P<0.05);Compared with the SAP group,the expression level of GP2 in the treatment group of Dai-Huang-Fu-Zi-Tang increased significantly(P<0.05).Conclusions: 1.20% L-arginine was injected intraperitoneally to establish SAP mouse model successfully.2.After the occurrence of SAP,the GP2 expression of intestinal M cell transporter is significantly reduced,suggesting that the M cell as the initiation of immune response isdamaged,the intestinal immune response cannot be started normally,and the intestinal immune barrier is damaged.3.After treatment with Dai-Huang-Fu-Zi-Tang,the expression of intestinal M cell transporter GP2 is significantly increased,indicating that the function of M cells to initiate intestinal immune response is restored,and the intestinal mucosal immune function is enhanced to prevent the intestinal flora and endotoxin from shifting and alleviating.The damage to the whole body caused by SAP;Dai-Huang-Fu-Zi-Tang has a very good role in promoting the recovery of intestinal function. |
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