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肠三叶因子和黏蛋白对烧伤血清所致肠上皮细胞增殖移行能力变化的影响
引用本文:王焕,吴修文,万千雪,金星,孙勇,吴丹,曾俊杰,彭曦. 肠三叶因子和黏蛋白对烧伤血清所致肠上皮细胞增殖移行能力变化的影响[J]. 中华烧伤杂志, 2011, 27(5). DOI: 10.3760/cma.j.issn.1009-2587.2011.05.006
作者姓名:王焕  吴修文  万千雪  金星  孙勇  吴丹  曾俊杰  彭曦
作者单位:1. 蚌埠医学院护理学院,233000
2. 400038 重庆,第三军医大学西南医院全军烧伤研究所,创伤、烧伤与复合伤国家重点实验室
基金项目:国家自然科学基金,“重大新药创制”科技重大专项,创伤、烧伤与复合伤国家重点实验室自主研究课题
摘    要:
目的 观察烧伤血清所致肠上皮细胞增殖、移行能力的变化,以及联合应用肠三叶因子(ITF)和黏蛋白对其的影响.方法 将大鼠小肠上皮细胞株IEC-6传代培养,按照随机数字袁法分为正常对照组、烧伤血清组、ITF+烧伤血清组、黏蛋白+烧伤血清组和ITF+黏蛋白+烧伤血清组,均采用DMEM培养液培养,其内分别添加体积分数10%小牛血清、体积分数10%烧伤大鼠血清、25 μg/mL ITF和体积分数10%烧伤大鼠血清、250 μg/mL黏蛋白和体积分数10%烧伤大鼠血清以及联合使用上述剂量ITF、黏蛋白和烧伤大鼠血清.处理后0~4d观察细胞增殖能力;划痕实验后12、24、36、48、72 h观察细胞移行情况;Transwell法(细胞置于上室、培养液加入下室)培养4、6、8、10、12 h,观察细胞变形能力,以下室内细胞计数表示.对数据进行t检验.结果 (1)细胞增殖能力.处理后1~4d烧伤血清组细胞数量明显低于正常对照组(t值为-16.569 ~ - 2.613.P<0.05或P<0.01).ITF+烧伤血清组和黏蛋白+烧伤血清组细胞数量与烧伤血清组接近(t值分别为0.037~0 740、0.116~0.429,P值均大于0 05);处理后2d,ITF+黏蛋白+烧伤血清组细胞数量明显高于烧伤血清组(t =6.484,P<0.01)、ITF+烧伤血清组(t =3.838,P<0.01).(2)细胞移行能力.烧伤血清组划痕后各时相点细胞移行距离均远远短于正常对照组(t值为-37.594~ -6.727,P值均小于0.0I).与烧伤血清组比较,黏蛋白+烧伤血清组划痕后各时相点细胞移行距离无明显变化(t值为0.055 ~0.589,P值均大于0 05).ITF+烧伤血清组划痕后12、24、36 h细胞移行距离分别为(47±6)、(126±13)、(170±11) μm,明显长于烧伤血清组[(42±7)、(98±14)、(154±22) μm,t值为2.230~4.817,P<0.05或P<0.01].ITF+黏蛋白+烧伤血清组划痕后各时相点细胞移行距离明显长于烧伤血清组(t值为2.982 ~7.390,P<0.05或P<0.01),划痕后12 ~48 h明显长于ITF+烧伤血清组(t值为2.707 ~2.918,P<0.05或P<0.01).(3)细胞变形能力.与正常对照组比较,烧伤血清组穿孔细胞数大幅减少(t值为- 23.965 ~ -6.436,P值均小于0.01).与烧伤血清组比较,黏蛋白+烧伤血清组各时相点穿孔细胞数改变不明显(t值为0.199~ 0.797,P值均大于0 05);ITF+烧伤血清组各时相点穿孔细胞数均明显增加(t值为3.650 ~10.028,P值均小于0.01).ITF+黏蛋白+烧伤血清组各时相点穿孔细胞数明显多于烧伤血清组(t值为4.313~15.100,P值均小于0.01),培养10、12 h时穿孔细胞数分别为(328±47)、(465±37)个,明显多于ITF+烧伤血清组[(277±25)、(353±34)个,t值分别为3.051、6 945,P值均小于0.01].结论 ITF对肠上皮细胞增殖影响有限,但能明显改善细胞变形能力,促进细胞移行;单独使用黏蛋白对细胞无明显作用,与ITF联合应用能增强ITF的作用.ITF维护肠黏膜屏障的主要机制为促进细胞移行.

关 键 词:烧伤  黏蛋白类  细胞增殖  细胞移行  肠三叶因子  肠上皮细胞

Effects of intestinal trefoil factor combined with mucin on ability of proliferation and migration of intestinal epithelial cells after being treated by burn rat serum
WANG Huan,WU Xiu-wen,WAN Qian-xue,JIN Xing,SUN Yong,WU Dan,CAO Jun-jie,PENG Xi. Effects of intestinal trefoil factor combined with mucin on ability of proliferation and migration of intestinal epithelial cells after being treated by burn rat serum[J]. Chinese journal of burns, 2011, 27(5). DOI: 10.3760/cma.j.issn.1009-2587.2011.05.006
Authors:WANG Huan  WU Xiu-wen  WAN Qian-xue  JIN Xing  SUN Yong  WU Dan  CAO Jun-jie  PENG Xi
Abstract:
Objective To observe the effect of intestinal trefoil factor (ITF) combined with mucin on the ability of proliferation and migration of intestinal epithelial cells ( IEC ) after being treated by burn rat serum.Methods The rat IEC-6 cell lines were subcultured and divided into control group ( C,cultured with DMEM medium containing 10% calf serum),burn serum group ( BS,cultured with DMEM medium containing 10% burn rat serum),burn serum + ITF group (B + I,cultured with DMEM medium containing 10% burn rat serum and 25 μg/mL ITF),burn serum + mucin group ( B + M,cultured with DMEM medium containing 10% burn rat serum and 250 μg/mL mucin),and burn serum + ITF + mucin group ( B + I + M,cultured with DMEM medium containing 10% burn rat serum,25 μg/mL ITF,and 250 μg/mL mucin) according to the random number table.Cells were counted on post culture day (PCD) 0,1,2,3,4,reflecting cell proliferation ability.Cell migration distance was measured at post scratch hour (PSH) 12,24,36,48,72.Then,cells of each group were placed in upper compartment of Transwell chamber while the corresponding medium was respectively added into lower compartment of Transwell chamber.Cells in lower compartment of Transwell chamber were counted at post culture hour (PCH) 4,6,8,10,12,reflecting cytomorphosis ability.Data were processed with t test.Results ( 1 ) Cell proliferation ability.The cell numbers in BS group on PCD 0,1,2,3,4 were significantly less than those in C group ( with t values from -16.569 to -2.613,P <0.05 or P < 0.01 ).The cell number showed no statistical difference between B + I and BS groups,and between B + M and BS groups at each time point ( with t values respectively from 0.037 to 0.740 and 0.116 to 0.429,P values all above 0.05 ).The cell number in B + I + M group on PCD 2 was respectively larger than that in BS group ( t =6.484,P <0.01 ) and B + I group ( t =3.838,P <0.01).(2) Cell migration distance in BS group at PSH 12,24,36,48,72 was significantly shorter than that in C group ( with t values from - 37.594 to - 6.727,P values all below 0.01 ).There was no obvious difference in cell migration distance between BS and B + M groups at each time point ( with t values from 0.055 to 0.589,P values all above 0.05).Cell migration distance in B + I group at PSH 12,24,36was respectively (47 ±6),( 126 ± 13),( 170 ± 11 ) μm,all longer than those in BS group [ (42 ± 7),(98 ±14),(154 ±22) μm,with t values from 2.230 to 4.817,P <0.05 or P <0.01].Cell migration distance in BS group at PSH 12,24,36,48,72 and B +I group at PSH 12,24,36,48 was respectively shorter than that in B + I + M group ( with t values respectively from 2.982 to 7.390 and 2.707 to 2.918,P <0.05 or P < 0.01 ).(3) Cytomorphosis ability.Compared with those of C group,cell counts in lower compartment of BS group at PCH 4,6,8,10,12 were significantly decreased (with t values from - 23.965 to -6.436,P values all below 0.01).Cell count in lower compartment of BS group at PCH 4,6,8,10,12 was respectively less than that of B + I group ( with t values from 3.650 to 10.028,P values all below 0.0l ) and similar to that of B + M group ( with t values from 0.199 to 0.797,P values all above 0.05 ).Cell counts in lower compartment of B +I + M group at PCH 4,6,8,10,12 were significantly larger than those of BS group (with t values from 4.313 to 15.100,P values all below 0.01 ).Cell count in lower compartment of B +I+M group at PCH 10 (328 ±47) and PCH 12 (465 ±37) was respectively larger than that in B +I group (277 ± 25,353 ± 34,with t value respectively 3.051,6.945,P values all below 0.01 ).Conclusions ITF can improve cytomorphosis ability for promoting cell migration with limited effect on cell proliferation,which can be enhanced with addition of mucin.The main mechanism of ITF in maintaining intestinal mucosal barrier may be attributed to acceleration of cell migration.
Keywords:Burns  Mucins  Cell proliferation  Cell migration  Intestinal trefoil factor  Intestinal epithelial cells
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