The therapeutic efficacy of mesenchymal stem cells on ileal pouchitis of rats
Objective To investigate the therapeutic effect and mechanism of human umbilical cord mesenchymal stem cells(hUC-MSCs)on rat model of ileal pouchitis after total colorectal resection and ileal pouch anal anastomosis(IPAA).Methods The ileal pouchitis models were established in 20 male Sprague-Dawley rats.After successful modeling,2 rats were randomly selected for stem cell tracing.The remaining 18 rats were randomly divided into MSC group,phosphate buffer salt solution group(PBS group)and dextrosan sulfate sodium group(DSS group)with 6 rats each.A total of 2 × 106 hUC-MSCs were intra-peritoneally injected once,and 1 ml PBS was intraperitoneally injected once,without special intervention.The changes of body weight,fecal score,blood stool status and disease activity index(DAI)score of each group were recorded,and the rats were killed after 4 days of administration.The bag samples were collect-ed for histopathological score,myeloperoxidase(MPO)expression level,interleukin(IL)-1β,IL-6,IL-10,tumor necrosis factor-α(TNF-α)mRNA expression and protein expression.The comparison between the two groups of normal distribution data was performed by t test.Mann-Whitney U rank sum test was used to compare the non-normal distribution data and rank data between the two groups.Kruskal-Wallis H-rank sum test was used to compare multiple independent samples.Results The concentration of MSCs in the storage bag was obvious after intraperitoneal injection,and the fluorescence disappeared at 4th day after in-jection.The weight in MSC group began to recover,and the symptoms of blood and stool disappeared.However,the body weight of rats in PBS group and DSS group slowed down but continued to decline,and the symptoms of blood and stool were not improved.After 4 days of stem cell therapy,DAI in MSC group was significantly lower than that in PBS group and DSS group(0.89±0.34 vs.2.95±0.25,Z=-2.966,P＜0.01;0.89±0.34 vs.2.78±0.17,Z=-2.956,P＜0.01).Histopathological scores in MSC group were significantly lower than those in PBS group and DSS group(1.50±0.84 vs.8.17±1.33,Z=-2.989,P＜0.01;1.50±0.84vs.8.33±1.51,Z=-2.956,P＜0.01).The MPO protein ex-pression was significantly decreased(0.02±0.00 vs.0.07±0.00,Z=-2.892,P＜0.01;0.02±0.00 vs.0.08±0.00,Z=-2.903,P＜0.01);The mRNA relative expression levels of IL-1β,IL-6 and TNF-α in MSC group were also significantly lower than those in PBS and DSS groups(0.02±0.00 vs.0.06± 0.02,0.02±0.00 vs.0.06±0.02,Z=-2.882,P＜0.01;0.05±0.01 vs.0.12±0.01,0.05± 0.01 vs.0.11±0.01,Z=-2.882,P＜0.01;0.04±0.00 vs.0.06±0.01,0.04±0.00vs.0.06± 0.01,Z=-2.882,P＜0.01);The expression levels of IL-1β,IL-6 and TNF-α protein(pg/mg)were also significantly decreased(0.73±0.04 vs.1.40±0.32,0.73±0.04 vs.1.41±0.36,Z=-2.882,P＜0.01;4.85±0.29 vs.6.68±0.24,4.85±0.29 vs.6.65±0.42,Z=-2.882,P＜0.01;2.39± 0.39 vs.4.14±0.25,2.39±0.39 vs.4.07±0.27,Z=-2.882,P＜0.01).The mRNA relative ex-pression and protein expression of IL-10 in MSC group were higher than those in PBS group and DSS group(0.10±0.01 vs.0.03±0.00,0.10±0.01 vs.0.03±0.00,Z=-2.882,P＜0.01;3.68±0.27 vs.2.38±0.28,3.68±0.27 vs.2.52±0.33,Z=-2.887,P＜0.01).Conclusion hUC-MSCs have therapeutic effect on IPAA postoperative ileal pouchitis in rats.The therapeutic effect of hUC-MSCs on ileal pouchitis may be alleviated by down-regulating the expression level of pro-inflammatory factors and up-regu-lating the expression level of anti-inflammatory factors.
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