Effect of adipose tissue derived mesenchymal stem cells via different routes of injection on rats with colitis

doi:10.3877/cma.j.issn.2095-1221.2017.05.005

Abstract

Abstract:

Objective

To investigate the effect of adipose tissue derived mesenchymal stem cells (ADMSCs) via intraperitoneal and intravenous injection on colitis induced by trinitrobenzenesulfonic acid (TNBS).

Methods

Totally 32 rats were randomly divided into a normal group, an intraperitoneal injection group, an intravenous injection group and a model group (n = 8). The animal model of inflammatory bowel disease (IBD) was induced by TNBS and ADMSCs were administrated intraperitoneally and intravenously. Disease activity index (DAI) were recorded, the microscopic changes and macroscopic damage of colon were observed, myeloperoxidase (MPO) activity were determined and the expression of Ki-67⁺cell were measured in colon. The expressions of IL-1βand TNF-αin the peripheral blood were compared and the expression level of gene (TGF-α, IL-6, IL-17A, IL-10) were determined in the colon. Single factor analysis of variance and independentttest were used for statistical analysis.

Results

Weight recovery in the intraperitoneal injection group 98.05±0.63 was higher than that in the intravenous injection group 94.32±0.48, (t= 12.281,P= 0.000) in day 6. At the same time, the DAI score in the intraperitoneal injection group 1.71±0.75 was significantly lower than that in intravenous injection group 3.57±0.97, (t= -3.980,P= 0.002). In addition, the MPO concentration of colon tissues in the intraperitoneal injection group (95.75±5.52) U/g was lower than that in the intravenous injection group (74.37±5.12)U/g, (t=-7.513,P= 0.000). The pathological score of colon in the intraperitoneal injection group 2.14±0.69 was lower than that in the intravenous injection group 3.57±0.76, (t= -3.612,P= 0.004). The immunofluorescence of the colonic tissues revealed more Ki67⁺cells in the intraperitoneal injection group than in the intravenous injection group. ELISA showed that the intraperitoneal injection group (130.71±7.08) pg/ml had lower concentration of IL-1βin plasma than did the intravenous injection group (163?±9.09) pg/ml, (t =-8.518,P= 0.000). The plasma concentration of TNF-αin the intraperitoneal injection group (201.71±6.75) pg/ml was also lower than that in the intravenous injection group (242.28±8.30) pg/ml, (t=-10.033,P= 0.000). In addition, the results of RT-qPCR in the colon tissues showed that the expression levels of IL-6 mRNA in the intraperitoneal injection group (4.34±0.48) were lower than those in the intravenous injection group (6.15±1.05,t=-4.147,P= 0.001) and the expression levels of IL-17A mRNA in intraperitoneal injection group 2.61±0.53 were also lower than those in the intravenous injection group (3.57±0.46,t= -4.301,P= 0.001). However, the expression levels of IL-10 mRNA in the intraperitoneal injection group (37.75±4.46) were higher than those in the intravenous injection group (27.68±2.25,t= 5.327,P?= 0.001) and the expression levels of TGF-βmRNA in the intraperitoneal injection group (15.82±0.99) were higher than those in the intravenous injection group (11.97±2.25,t= 3.740,P= 0.003).

Conclusion

Intraperitoneal injection of ADMSCs is better than intravenous injection, which may be a better route of administration for ADMSCs treating IBD.

Key words: Inflammatory bowel disease, Adipose tissue derived mesenchymal stem cells, Routes of injection

Zhengwei Fu, Zhenyu Zhang, Haiyan Ge. Effect of adipose tissue derived mesenchymal stem cells via different routes of injection on rats with colitis[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2017, 07(05): 277-286.

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Figures/Tables 11

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分值	相对体重下降（﹪）	大便隐血	大便性状
0	< 1	阴性	正常
1	1 ~ 5	隐血阳性	软便
2	5 ~ 10	?	大便松散
3	10 ~ 15	?	稀糊样便
4	> 20	肉眼血便	水样腹泻

分值	相对体重下降（﹪）	大便隐血	大便性状
0	< 1	阴性	正常
1	1 ~ 5	隐血阳性	软便
2	5 ~ 10	?	大便松散
3	10 ~ 15	?	稀糊样便
4	> 20	肉眼血便	水样腹泻

基因	正向引物（5'-3'）	反向引物（5'-3'）
IL-17A	AAGCACAGAAAGCATGATCCG	GAGTCCAGGGTGAAGTGGA
IL-10	CCTTCCTCCGTGTGGTTTGA	TTCTTGAGGGCCACTTCGTA
IL-6	GCCAGATAGAGTCGTTGCCC	AGCTAGTTGCCGTGTGTCTG
TGF-β	CCCACATGAGATCAGCCTCC	GGAGTTGGGTGGCAAGAGTT
GAPDH	TCAATGAAGGGGTCGTTGAT	CGTCCCGTAGACAAAATGGT

基因	正向引物（5'-3'）	反向引物（5'-3'）
IL-17A	AAGCACAGAAAGCATGATCCG	GAGTCCAGGGTGAAGTGGA
IL-10	CCTTCCTCCGTGTGGTTTGA	TTCTTGAGGGCCACTTCGTA
IL-6	GCCAGATAGAGTCGTTGCCC	AGCTAGTTGCCGTGTGTCTG
TGF-β	CCCACATGAGATCAGCCTCC	GGAGTTGGGTGGCAAGAGTT
GAPDH	TCAATGAAGGGGTCGTTGAT	CGTCCCGTAGACAAAATGGT

分组	动物数	第1天	第2天	第3天	第4天	第5天	第6天
正常组	8	101.00±0.84	102.07±0.77	102.74±0.64	103.08±0.40	103.37±0.44	103.74±0.33
腹腔注射组	8	96.80±0.90	94.81±0.80	93.27±0.46	5.22±0.51	96.24±0.56	98.05±0.63
静脉注射组	8	97.55±1.33	93.18±0.53	92.14±0.62	92.70±0.53^a	93.47±0.48^a	94.32±0.48^a
模型组	8	97.95±1.00	92.10±0.67	89.88±0.78	90.67±0.71^a	91.41±0.51^a	92.37±0.45^a
F值	?	22.145	283.773	548.216	680.569	751.259	727.073
P值	?	0.186	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

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