烫伤血清对大鼠结肠平滑肌细胞骨架的影响

目的 了解烫伤血清对体外培养的大鼠结肠平滑肌细胞骨架的影响,初步探讨伤后胃肠动力障碍的可能机制. 方法 分离培养健康成年Wistar大鼠结肠平滑肌细胞.另取2只健康成年Wistar大鼠,其中1只不致伤,另一只致深Ⅱ度烫伤,采集2只大鼠血液分离血清,用培养液配成体积分数20%的烫伤血清和体积分数20%的正常血清.采用随机数字表法将大鼠平滑肌细胞分为烫伤血清组和正常血清组,分别用2种血清培养.于处理1、3、6、12 h时(每组每时相点样本数均为10),用流式细胞仪检测细胞肌动蛋白表达水平,并采用蛋白质印迹法检测β微管蛋白的相对含量.对数据行t检验. 结果 (1)烫伤血清组细胞处理1、3、6、12 h时,肌动蛋白荧光强度分别为59±4、26±6、39±6、42±6,正常血清组分别为95±10、91±10、102±9、97±9,组间比较,差异均有统计学意义(t值分别为10.528、18.069、18.748、16.647,P＜0.05或P＜0.01);烫伤血清组组内比较,处理3 h时降至最低点,6、12 h持续回升.(2)烫伤血清组处理1、3、6、12 h时,β微管蛋白的相对含量分别为14.44±0.26、8.61±0.19、11.76±0.31、12.13±0.29;正常血清组分别为22.37±1.15、21.87±1.79、23.24±1.55、21.99±2.02,组间比较差异有统计学意义(t值分别为21.176、23.365、23.000、15.273,P值均小于0.01);烫伤血清组组内比较,处理3 h时达最低,6、12 h有所回升但势头缓慢.结论 烫伤血清作用初始阶段,可使体外培养的大鼠肠道平滑肌细胞骨架含量减少,随后细胞骨架含量呈回升趋势,这可能与平滑肌细胞对烫伤血清刺激的耐受、适应和自我修复有关.

Influence of serum from rats with scald on the cytoskeleton of colonic smooth muscle cells of rats

Objective To study the influence of serum from scalded rats on the cytoskeleton of colonic smooth muscle cells (CSMC) of rats cultured in vitro, and to probe the possible mechanism of gastrointestinal motility disorder after burn. Methods CSMC isolated from healthy adult Wistar rat were cultured and divided into scald serum group (SS) and normal serum group (NS) according to the random number talbe. Two normal Wistar rats were used, one of which was inflicted with deep partial-thickness scald. Serum was obtained from blood collected from these two rats respectively and diluted to 20% in concentration. Serum from scald and normal rats were respectively added to the culture of CSMC in SS and NS groups. The expression of actin and the relative content of β-tubulin in CSMC was respectively determined with flow cytometry and Western blot at post treatment hour (PTH) 1, 3, 6, and 12 (with 10 samples in each group at each time point). Data were processed with t test. Results Fluorescence intensity of actin in SS group at PTH 1, 3, 6, and 12 was respectively 59 ± 4, 26 ± 6, 39 ± 6, and 42 ± 6, all significantly lower than those in NS group (95 ± 10, 91 ± 10, 102 ± 9, and 97 ± 9, with t value respectively 10.528, 18. 069, 18. 748, 16. 647,P ＜0.05 orP ＜0.01 ). In SS group, the fluorescence intensity decreased to the nadir at PTH 3, and then increased persistently at PTH 6 and 12. (2) Relative content of β-tubulin in SS group at PTH 1,3, 6, and 12 was respectively 14.44 ± 0.26, 8.61 ± 0. 19, 11.76 ± 0.31, and 12. 13 ± 0.29, all significantly less than those in NS group (22.37 ± 1.15, 21.87 ± 1.79, 23.24 ± 1.55, and 21.99 ± 2.02, with t value respectively 21. 176, 23. 365, 23. 000, 15. 273, P values all below 0. 01 ). In SS group, the relative content of β-tubulindecreased to the nadir at PTH 3 and increased slowly at PTH 6 and 12. Conclusions The reduction of CMSC content which has the tendency of increasing later, can be attributed to the influence of scald serum in initial stage. This may be related to the tolerance and adaptation to scald serum and self-repair of CMSC.