The Experimental Study Of Tripterygium Wilfordii Polyglycosidium In The Treatment Of Rat Pouchitis

Pouchitis is the most frequent forward complication after colorectal resection and ileal pouch anal anastomosis among patients with ulcerative colitis, it seriously affect the postoperative life quality of those patients, severe cases make the pouch stand by, and even need permanent ileostomy surgery. The etiology of pouchitis remains unclear, abnormal immune regulation plays an important role in the pathogenesis. Immune inhibitor Tripterygium wilfordii polyglycosidium perform great anti-inflammatory and immunosuppressive action, and has been widely used in clinical practice with satisfactory curative effect. This topic explored the intervention of Tripterygium wilfordii polyglycosidium on pouchitis via stabilized rats ileal pouchitis model building and explore the mechanism, aim at researching novel strategy of pouchitis treatment.Part1 The modeling of Rat PouchitisObjective: We build rat IPAA model by microsurgery and iduce rats pouchitis by DSS separately. Methods: Microsurgery was used to establish IPAA model, the model rats were randomly divided into Sham, IPAA-NI(non-intervention, NI) and IPAA-DSS group(4% DSS solution 4 days after 31 days postoperative). During the experiment records each rat body weight change, bloody stool situation, excrement characters(score). Taking terminal ileum tissue from Sham rats, and pouch from IPAA-NI and IPAA-DSS rats for histologic grading; Real-time fluorescent quantitative PCR detected IL-1?, IL-6, IL-10 and TNF-? m RNA expression level. Results: The postoperative rats' weight reached to lowest level about(280.68±16.07) g at(8.83±1.47) days; IPAA-DSS rats' weight lose(59.50±7.23)g after with DSS, compared with IPAA-NI groups was statistically significant difference(P < 0.05). All rats in IPAA-DSS group had bloody stool(6/6), Sham and IPAA-NI group had no bloody stool, IPAA-DSS fecal scores were inferior to Sham and IPAA-NI group, and IPAA-NI group were inferior to Sham group(P < 0.01); The histological scores in IPAA-DSS were higher than SHAM and IPAA-NI group(P < 0.01), and scores in IPAA-DSS were greater than Sham and IPAA-NI group(P < 0.01), and IPAA-NI group were greater than Sham group(P < 0.01). There were more IL-1?, IL-6, and TNF-? m RNA expression(all P < 0.01) and less IL-10(P < 0.01) in IPAA-DSS compared to other two groups.Part 2 The therapeutic effect of Tripterygium wilfordii polyglycosidium in Rat PouchitisObjective: To explore the therapeutic effect of Tripterygium wilfordii polyglycosidium on rat pouchitis and find the possible mechanism. Methods: The rats pouchitis model were randomly divided into no intervention groups(NI), normal saline(NS), Tripterygium wilfordii polyglycosidium(TWP), and were separately given 3 ml saline, 12mg/(kg*d)Tripterygium wilfordii polyglycosidium by lavaging 7 days. During the experiment records each rat body weight change, bloody stool situation, excrement characters(score). Taking pouch for histologic grading at the eighth day; Real-time fluorescent quantitative PCR detected IL-1?, IL-6, IL-10 and TNF-? m RNA expression level. Immunohistochemistry method was used to measure occludin protein.Saving blood to test liver function. Results: Rats in TWP loss less weight than that of NI and NS groups. Gross blood stool was disappeared on day(6.00±0.89) in NI group,(5.83±0.75) in NS group,(3.33±0.52) in TWP group. Fecal scores were obviously improved in TWP group, higher than that of NI and NS groups(P < 0.01). Pouch histopathologic scores were lower in TWP group compared to other two groups(P < 0.01). There were less IL-1?, IL-6, and TNF-? m RNA expression(all P< 0.01) and more IL-10(P < 0.01) in TWP compared to other two groups. Occludin expression level was higher in TWP than that of NI and NS groups(P < 0.05). The ALT and AST in TWP treatment group had no significant difference(P>0.05).Conclusion1. Successfully producing the rat model of ileal pouch by microsurgery and inducing the rat model of pouchitis with DSS.2. The relative increase of pro-inflammatory factors and the relative decrease of anti-inflammatory factors promote the damage of pouch.3. TWP can treat rat pouchitis successfully. TWP may treat rat pouchitis by decrease pro-inflammatory factors IL-1?, IL-6 and TNF-?,and increase the relative of anti-inflammatory factors IL-10 promote the recover of intestinal barrier.4. The clinical dose TWP has no obvious impact on ALT and AST of expernmental rats, it provides experimental basis for the clinical safe medication.