| Objectives:To observe the effects of Fecal Microbiota Transplantation(FMT)on the colonic pathological histology,mucosal Th1/Th2 cells,and related cytokine level in colonic tissue.And then to investigate the possible immunological mechanisms of FMT in mice with colitis induced by dextran sulfate sodium(DSS).Methods:Forty-eight C57BL/6 male mice were randomly divided into intervention group,control group and feces donor group with a ratio of 3:2:1.Twenty-four mice in the intervention group were drinked with 2% DSS solution for 5 days,and were subdivided into group A(FMT),group B(Etiasa),group C(Saline)randomly at the 6th day.The mice in group A were received fecal filtrate as an enema(500 mg/ml,0.3ml/times),in group B received Etiasa suspension liquid(25 mg/ml,0.3 ml/times),and in group C received saline(0.3 ml/times)respectively,two times per day for five consecutive days.The mice in control group were divided into group D(Acute colitis)and group E(Normal),which were drinked with 2% DSS solution or distilled water respectively for 7 days,and then were executed at day 8th day.The fresh faeces from mice in group F(Faeces donors)were collected for fecal filtration.The general healthy,body weight,disease activity index,spleen index,the colon length and colonic histopathological index were evaluated.The expression of Gata-3/T-bet positive cells in colon were determined by immunohistochemical method.And the colonic concentration of TNF-?,IFN-?,IL-4,and IL-10 were detected by enzyme-linked immunosorbent assay(ELISA).Results:(1)FMT significantly increased the spleen index,and improved the DAI score,colon length,and colonic pathological score in mice with DSS-induced colitis.(1)The spleen index(mg/ml)was 5.25±1.11 in group A(FMT),4.61±0.60 in group B(Etiasa),2.26 ± 0.33 in group C(Saline),3.36 ± 0.40 in group D(Acute Colitis),and3.00±0.17 in group E(Normal)respectively.It was significantly higher either in group A or in group B than in group D(PAD=0.015;PBD=0.003),however there was no significant difference between group A and group B(PAB=0.860).(2)The DAI score in group A(FMT,5.75±1.58)was significantly lower than in group C(Saline,8.63±1.30)(PAC=0.000),however there was no significant difference between group A and group B(Etiasa,7.00±1.07)(PAB=0.075).(3)The colon length(cm)was 7.65±0.89 in group A(FMT)?6.86±0.66 in group B(Etiasa)?6.13±0.37 in group C(Saline),5.79±0.41 in group D(Acute Colitis),and8.50±0.77 in group E(Normal)respectively.It showed a significant difference between group A and group B(PAB=0.021).(4)The colonic histopathological index was 3.88±0.83 in group A(FMT),6.13±1.46 in group B(Etiasa)? 7.37 ± 1.51 in group C(Saline),10.37 ± 0.92 in group D(Acute Colitis),and 0.00±0.00 in group E(Normal)respectively.The index in group A was significantly lower than in group B(PAB=0.029),however,there was no significant difference between group B and group C(PBC=0.701).(2)FMT increased T-bet and Gata-3 positive cell infiltration in colon tissues,and moved the imbalance of T-bet/Gata-3 to right in mice with DSS induced colitis.The colonic Gata-3 positive cell numbers in group A(FMT)were more than either in group B(Etiasa)or in group C(Saline)(P<0.005).Although the T-bet positive cell numbers in group A were more than either in group B or in group C,while there were no significantly different(P>0.005).T-bet/Gata-3 ? 1 in group A was 25%,which was significantly lower than in group C(100%,PAC=0.003),while similar to that in group B(37.5%,PAB=0.602).?FMT significantly decreased the expressions of TNF-?,IFN-?,IL-4,and IL-10 in the colon tissue in DSS-induced colitis mice.(1)The mucosal concentration of TNF-?(pg/ml)was 5324.38 ± 813.86 in group A(FMT),5471.25±528.40 in group B(Etiasa),4615.88±569.07 in group C(Saline)?6657.13±889.14 in group D(Acute Colitis),4196.13±623.66 in group E(Normal)respectively.(2)The mucosal concentration of IFN-?(pg/ml)was 5511.25 ± 672.53 in group A,5010.00±413.21 in group B,5338.75±614.53 in group C,7327.50±1132.18 in group D,5531.25±642.06 in group E respectively.(3)The mucosal concentration of IL-4(pg/ml)was 563.25 ± 35.30 in group A,562.50 ± 35.82 in group B,498.38 ± 50.53 in group C,652.00 ± 46.71 in group D,516.63±64.92 in group E respectively.(4)The mucosal concentration of IL-10(pg/ml)was 2462.35 ± 247.03 in group A,2316.25±199.35 in group B,2374.88±240.61 in group C,3067.38±198.37 in group D,2314.88±288.58 in group E respectively.The mucosal concentrations of four cytokines in group A,group B,or in group C were significantly lower than those in group D(P<0.05)respectively,while the levels of TNF-?,IFN-?,or IL-10 were no significantly different among the group A,group B,and group C(P>0.05).However,the IL-4 concentration in group A was higher than in group C(P=0.01)and there was no significantly different between group A and group B(P=0.975).Conclusion: FMT could balance the colon mucosal Th1 / Th2 cells,and reduce the colonic levels of IL-4,IL-10,TNF-? and IFN-?,which contributed to improve the colonic histologial index in the C57BL/6 Mice with DSS-induced colitis. |
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