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纸质出版日期:2015
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朱磊, 沈洪, 刘丽, 等. 健脾补肾清肠化湿方对静脉移植MSCs向溃疡性结肠炎大鼠结肠迁移分化的影响[J]. 中国实验方剂学杂志, 2015,21(21):88-92.
ZHU Lei, SHEN Hong, LIU Li, et al. Effects of Jianpi Bushen Qingchang Huashi Decoction on Migration and Differentiation of MSCs in Ulcerative Colitis Rat[J]. Chinese journal of experimental traditional medical formulae, 2015, 21(21): 88-92.
朱磊, 沈洪, 刘丽, 等. 健脾补肾清肠化湿方对静脉移植MSCs向溃疡性结肠炎大鼠结肠迁移分化的影响[J]. 中国实验方剂学杂志, 2015,21(21):88-92. DOI: 10.13422/j.cnki.syfjx.2015210088.
ZHU Lei, SHEN Hong, LIU Li, et al. Effects of Jianpi Bushen Qingchang Huashi Decoction on Migration and Differentiation of MSCs in Ulcerative Colitis Rat[J]. Chinese journal of experimental traditional medical formulae, 2015, 21(21): 88-92. DOI: 10.13422/j.cnki.syfjx.2015210088.
目的: 观察健脾补肾清肠化湿方对静脉移植的大鼠骨髓间充质干细胞(MSCs)向溃疡性结肠炎模型大鼠结肠迁移归巢的影响。方法: 体外分离培养SD大鼠MSCs
传至第3代
制备细胞悬液
加入0.78 mg·L-1健脾补肾清肠化湿方共培养
并以4'
6-二脒基-2-苯基吲哚(4'
6-diamidino-2-phenylindole
DAPI)荧光标记备用。大鼠分为空白组
模型组
MSCs组
干预MSCs组和联合组。2
4
6三硝基苯磺酸(TNBS)法建立溃疡性结肠炎大鼠模型
空白组、模型组大鼠尾静脉分别iv 1 mL生理盐水
MSCs组大鼠
iv 1 mL MSCs(1×106个)
干预MSCs和联合组大鼠分别尾静脉iv体外中药干预的MSCs(1×106个)
联合组再予健脾补肾清肠化湿方13.6 g·kg-1
ig 10 d。第5
10天每组各处死5只大鼠
结肠标本进行大体形态和组织学评估
共聚焦显微镜观察各组DAPI标记MSCs在结肠内迁移情况
免疫组化检测肠干细胞标记物神经细胞RNA结合蛋白(Musashi-1)的表达。结果: 与正常组比较
模型组大鼠结肠可见广泛充血水肿
伴糜烂
肉眼可见溃疡形成
提示造模成功。除模型组和正常组外
各移植组结肠组织均能检测到DAPI标记的MSCs
且数量随时间的延长而逐渐增加;各治疗组均能改善大鼠结肠组织大体形态和组织病理
且第10天联合组优于MSCs组(P<0.05);各移植组能提高Musashi-1的表达
第10天联合组和干预MSCs组优于MSCs组(P<0.05)。结论: 健脾补肾清肠化湿方能促进MSCs向结肠溃疡部位的迁移和分化
并能修复受损的结肠黏膜。
Objective: To observe whether Jianpi Bushen Qingchang Huashi decoction(JBQHD) can promote the migration of mesenchymal stem cells (MSCs) intravenously transplanted in rat's bone to the colon of ulcerative colitis (UC) rat model. Method: MSCs were isolated and cultured in vitro
then spread to the third generation to prepare cell suspension
and 0.78 mg·L-1 JBQHD was added into it for co-culture. Fluorescent pre-labeling by 4'
6-diamidino-2-phenylindole (DAPI) was carried out in vitro. The rats were divided into blank group
model group
MSCs group
intervened MSCs group and combined group. UC rats models were established by TNBS method. The rats of blank group and model group received intravenous injection with 1 mL normal saline respectively through tail veins. The rats of MSCs group received intravenous injection with 1 mL MSCs through tail veins(1×106 CFU).The rats of intervened MSCs group and combined group received intravenous injection with MSCs that had been intervened in vitro with the decoction(1×106 CFU)
and the combined group also received 13.6 g·kg-1JBQHD by ig for 10 days. Five rats were killed respectively in each group on Day 5 and Day 10.Gross morphological and histological evaluation was taken for the colon specimens
and confocal microscope was used to observe the migration of MSCs labeled by DAPI in the colon tissues;immunohistochemical staining was conducted to detect the expression of Musashi-1(a marker of intestinal stem cells). Result: Compared with the normal group
the rats in model group had extensive hyperemia and edema in colon tissues
accompanied by erosion and visible ulceration
indicating successful modeling. Except model group and normal group
MSCs labeled by DAPI can be detected in colon tissues in all other groups
and the quantity increased gradually with time.The all treatment groups could improve gross morphology and histopathology in colon tissues of rats
and the result in combined group was superior to that in MSCs group (P<0.05) on Day 10
and various transplantgroups could increase the expression of Musashi-1
and the result in combined group and intervened MSCs group was superior to that in MSCs group (P<0.05) on Day 10. Conclusion: JBQHD could not only promote the migration and differentiation of MSCs to the colon ulcers
but also repair the damaged colon mucosa.
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