严重烧伤对幼鼠生长发育的影响及重组人生长激素的治疗作用

发布时间：2018-04-11 08:26

本文选题：烧伤 + 幼鼠　；参考：《重庆医科大学》2013年硕士论文

【摘要】：第一部分严重烧伤对幼鼠生长发育及生长激素水平影响的实验研究目的观察幼鼠严重烧伤后体格发育、学习记忆能力和运动能力的改变，以及体内生长激素水平的变化趋势，为严重烧伤后干预性治疗打下基础。方法 3-4周Balb/c小鼠随机平均分成四组：空白对照组、阴性对照组、7秒烧伤组和10秒烧伤组，每组26只。烧伤前测量小鼠体重、体长、尾长，伤后1d、3d、7d时每组分别随机抽取6只小鼠采血， ELISA方法测定血清中生长激素含量。每组剩余的8只小鼠饲养至成年后再次测量小鼠的体重、体长、尾长，行水迷宫实验（包括定位航行实验和空间探索实验）测试学习记忆能力，行负重游泳实验和爬杆实验测试运动能力。结果（1）7s烧伤组和10s烧伤组小鼠的体重、体长、尾长增长百分比均落后于正常对照组（P＜0.05，，P＜0.01，P＜0.01）；（2）7s烧伤组小鼠的逃避潜伏期、跨越平台次数及目标现象时间比例与正常对照组无明显差异；10s烧伤组的逃避潜伏期明显长于正常对照组（P＜0.05），而空间探索实验示跨越平台次数和目标现象时间比例较正常对照组增加（P＜0.05， P＜0.01）。（3）7s烧伤组小鼠爬杆时间和正常对照组相比，差异无统计学意义，而负重游泳时间较正常对照组缩短；10s烧伤组小鼠的爬杆时间和负重游泳时间较正常对照组缩短（P＜0.05， P＜0.01）。（4）7s烧伤组小鼠伤后第1d、3d生长激素水平较正常对照组降低（P＜0.05），伤后7d时与正常对照组相比差异无统计学意义；10s烧伤组小鼠生长激素在伤后1d、3d和7d均低于对照组，（P＜0.01、P＜0.01、P＜0.05）。结论严重烧伤导致幼鼠体格发育不良，学习记忆能力和运动能力受到影响；同时严重烧伤后幼鼠体内生长激素水平明显降低。第二部分重组人生长激素对严重烧伤幼鼠的治疗作用目的观察rhGH对严重烧伤幼鼠体格发育、学习记忆能力、运动能力、血清白蛋白和症因子水平等的影响。方法 3-4周Balb/c小鼠，随机分成6组：正常对照组组、阴性对照组、10s烧伤组、NS对照组，rhGH1mg.kg-1.d-1治疗组和rhGH3mg.kg-1.d-1治疗组。造模前测量各组小鼠体重、体长和尾长。正常对照组小鼠不做处理；阴性对照组小鼠麻醉、脱毛等处理同烧伤组；10s烧伤组采用如上介绍方法制作10s烧伤模型；NS对照组、rhGH1mg.kg-1.d-1治疗组和rhGH3mg.kg-1.d-1治疗组烧伤后每日早上8点分别皮下注射NS、rhGH1mg/kg和rhGH3mg/kg，持续10d。伤后1d、3d、5d随机抽取正常对照组组、阴性对照组和10s烧伤组每组6只小鼠取血，伤后3d、5d随机抽取NS对照组，rhGH1mg.kg-1.d-1治疗组和rhGH3mg.kg-1.d-1治疗组每组6只小鼠取血，剩余小鼠饲养至成年,成年后再次测量体重、体长、尾长，行水迷宫实验（包括定位航行实验和空间探索实验）测试学习记忆能力，行负重游泳实验和爬杆实验测试运动能力。ELISA方法检测血清IL-6和TNF-α含量，同时检测血清中白蛋白、肌酐、葡萄糖、钙离子水平。结果（1）与10s烧伤组及NS对照组相比，rhGH1mg.kg-1.d-1治疗组小鼠体重、体长、尾长增长百分比增加，但差异无统计学意义；而rhGH3mg.kg-1.d-1治疗组的体重、体长、尾长增长百分比与10s烧伤组和NS对照组相比差异有统计学意义（P＜0.05，P＜0.05，P＜0.01）；（2）rhGH1mg.kg-1.d-1治疗组小鼠第6d逃避潜伏期较10s烧伤组和NS对照组缩短（P＜0.05），目标现象停留时间延长（P＜0.01）；rhGH3mg.kg-1.d-1治疗组小鼠第5d、第6d的逃避潜伏期较10s烧伤组和NS对照组均缩短（P＜0.05，P＜0.01），穿越平台次数和目标象限停留时间较烧伤组增多（P＜0.05，P＜0.01）；（3）rhGH3mg.kg-1.d-1治疗组小鼠负重游泳时间和爬杆时间较10s烧伤组及NS对照组延长，差异有统计学意义（P＜0.05）; （4）小鼠烧伤后血清白蛋白水平降低（伤后1d、3d、5d P＜0.01）；血肌酐水平升高（伤后1d、3d、5d P＜0.05）；血清钙降低（伤后1d、3d、5d P＜0.01）；血糖升高（3d、5d P＜0.05）；（5）伤后3d（治疗2d后），rhGH1mg.kg-1.d-1治疗组和rhGH3mg.kg-1.d-1治疗组白蛋白水平升高（P＜0.05）；rhGH1mg.kg-1.d-1治疗组和rhGH3mg.kg-1.d-1治疗组血清肌酐降低（P＜0.05）； rhGH3mg.kg-1.d-1治疗组血钙升高（P＜0.05）；rhGH1mg.kg-1.d-1治疗组和rhGH3mg.kg-1.d-1治疗组血糖降低（P＜0.05）。（6）伤后5d（治疗4d后），rhGH1mg.kg-1.d-1治疗组和rhGH3mg.kg-1.d-1治疗组白蛋白水平升高（P＜0.05）；rhGH1mg.kg-1.d-1治疗组和rhGH3mg.kg-1.d-1治疗组血清肌酐降低（P＜0.05）；rhGH1mg.kg-1.d-1治疗组和rhGH3mg.kg-1.d-1治疗组血糖降低（P＜0.05）；治疗组和烧伤组及NS对照组血钙水平无明显差异。（7）与正常小鼠比较，小鼠烧伤后1d、3d和5d血清中TNF-α和IL-6水平明显升高（P＜0.01）；rhGH1mg.kg-1.d-1治疗组2d、4d时IL-6浓度较烧伤组及NS对照组降低（P＜0.05）；rhGH3mg.kg-1.d-1治疗组在治疗后2d、4d降低（P＜0.01，P＜0.05）。rhGH1mg.kg-1.d-1治疗组和rhGH3mg.kg-1.d-1治疗组小鼠TNF-α水平在治疗后2d、4d与烧伤组及NS对照组无明显差异。结论幼鼠严重烧伤后，rhGH通过促进蛋白质合成，抑制炎症反应等，降低严重烧伤后的高分解代谢状态及炎症反应状态，从而改善严重烧伤引起的体格发育落后、学习记忆能力落后及运动能力下降。
[Abstract]:Experimental study on the effects of severe burn on growth and growth hormone level in young rats
objective
We observed the changes of physical development, learning and memory ability and motor ability, and the change trend of growth hormone levels in severely burned rats, which laid the foundation for intervention treatment after severe burn.
Method
The 3-4 week Balb/c mice were randomly divided into four groups: blank control group, negative control group, 7 seconds and 10 seconds burn group and burn group, 26 rats in each group. Burn measured before the mice weight, body length, tail length, 1D after injury, 3D, 7d in each group were randomly selected 6 mice blood, ELISA method growth hormone content in serum of each group. The remaining 8 mice reared to adulthood measured again after mice body weight, body length, tail length, water maze test (including navigation experiment and space exploration experiment) to test the ability of learning and memory for swimming experiment and pole test exercise capacity.
Result
(1) the percentage of body weight, body length and tail length in the 7S burn group and the 10s burn group were all behind the normal control group (P < 0.05, P < 0.01, P < 0.01).
(2) 7S burn mice escape latency, the number of platform across time and target phenomenon ratio had no significant difference with normal control group; 10s burn group the escape latency was significantly longer than the normal control group (P < 0.05), while the space exploration experiment times of crossing the platform and target as the proportion of time increased compared with normal control group (P < 0.05, P < 0.01).
(3) there was no significant difference in the climbing time between 7S burn group and normal control group, while the time of loading swimming was shorter than that of normal control group. The time of creeping rod and weight swimming in 10s burn group was shorter than that in normal control group (P < 0.05, P < 0.01).
(4) in 7S burn group, the level of growth hormone at 1D and 3D after injury was lower than that in normal control group (P < 0.05). There was no significant difference in 7d after injury compared with that in normal control group. The growth hormone in 10s burn group was lower than that in control group after injury (P < 0.01, P < 0.01, 7d < 0.05).
conclusion
Severe burn resulted in poor physical development in young rats, and learning memory and exercise ability were affected. Meanwhile, the growth hormone level in young rats after severe burn was significantly reduced.
The therapeutic effect of recombinant human growth hormone on young rats with severe burn injury in the second part
objective
The effects of rhGH on the physical development, learning and memory ability, exercise ability, serum albumin and the level of the disease factor in young rats with severe burn were observed.
Method
The 3-4 week Balb/c mice were randomly divided into 6 groups: normal control group, negative control group, 10s burn group, NS control group, rhGH1mg.kg-1.d-1 treatment group and rhGH3mg.kg-1.d-1 treatment group. Rats measured before the mice weight, body length and tail length. Mice in normal control group without treatment; negative control group mice were anesthetized. With the hair removal and burn group; 10s group adopts the method of making the burn burn 10s model; NS control group, rhGH1mg.kg-1.d-1 treatment group and rhGH3mg.kg-1.d-1 treatment group after burn at 8 every morning were subcutaneous injections of NS, rhGH1mg/kg and rhGH3mg/kg, 10d. after injury 1D, 3D, 5D were randomly selected from the normal control group, negative control group 10s and burn group 6 mice in each group blood 3D after injury, 5D were randomly selected from the NS control group, rhGH1mg.kg-1.d-1 treatment group and rhGH3mg.kg-1.d-1 treatment group, each group of 6 mice blood, the remaining mice were reared to adult, adult To measure body weight, body length, tail length, water maze test (including navigation experiment and space exploration experiment) to test the ability of learning and memory for swimming experiment and pole test exercise capacity.ELISA method for detection of serum IL-6 and TNF- alpha content, serum albumin, creatinine, glucose, calcium level.
Result
(1) compared with 10s burn group and NS control group, rhGH1mg.kg-1.d-1 treatment group, the body weight, body length, tail length growth percentage increased, but the difference was not statistically significant; while rhGH3mg.kg-1.d-1 treatment group's body weight, body length, tail length and percentage growth compared to 10s burn group and NS control group had significant difference (P < 0.05, P < 0.05, P < 0.01);
(2) rhGH1mg.kg-1.d-1 treatment group at 6D latency than 10s burn group and NS control group (P < 0.05), shorten the residence time of target phenomenon (P < 0.01); rhGH3mg.kg-1.d-1 treatment group at 5D, the 6D latency than 10s burn group and NS control group were shorter (P < 0.05, P < 0.01), the number of cross platform and target quadrant time than burn group increased (P < 0.05, P < 0.01);
(3) the time of swimming and the time of climbing pole in the rhGH3mg.kg-1.d-1 treatment group were longer than those in the 10s group and the NS control group, and the difference was statistically significant (P < 0.05).
(4) the level of serum albumin in burned mice decreased (1D, 3D, 5D P < 0.01), serum creatinine level increased (1D after injury, 3D, 5D P < 0.05), serum calcium decreased (1D, 3D, 5D 5D < 0.01) and blood sugar increased (< 0.05).
(5) 3D after injury (2D after treatment), rhGH1mg.kg-1.d-1 treatment group and rhGH3mg.kg-1.d-1 treatment group increased albumin level (P < 0.05); rhGH1mg.kg-1.d-1 treatment group and rhGH3mg.kg-1.d-1 treatment group serum creatinine decreased (P < 0.05); rhGH3mg.kg-1.d-1 treatment group, serum calcium (P < 0.05); rhGH1mg.kg-1.d-1 treatment group and rhGH3mg.kg-1.d-1 treatment group decreased blood glucose (P < 0.05).
(6) 5D after injury (4D after treatment), rhGH1mg.kg-1.d-1 treatment group and rhGH3mg.kg-1.d-1 treatment group increased albumin level (P < 0.05); rhGH1mg.kg-1.d-1 treatment group and rhGH3mg.kg-1.d-1 treatment group serum creatinine decreased (P < 0.05); rhGH1mg.kg-1.d-1 treatment group and rhGH3mg.kg-1.d-1 treatment group decreased blood glucose (P < 0.05); treatment group and burn group NS and control group serum calcium levels were not significantly different.
(7) compared with the normal mice, mice after burn 1D, TNF- alpha and IL-6 levels of 3D and 5D in serum were significantly increased (P < 0.01); rhGH1mg.kg-1.d-1 group 2D, 4D IL-6 concentration is lower than burn group and NS control group (P < 0.05); rhGH3mg.kg-1.d-1 treatment group after treatment, 2D, 4D decreased (P < 0.01, P < 0.05) in.RhGH1mg.kg-1.d-1 group and rhGH3mg.kg-1.d-1 group were TNF- levels at 2D after treatment, no significant difference between 4D and burn group and NS control group.
conclusion
After severe burn, rhGH can reduce the state of catabolism and inflammatory reaction after severe burn by promoting protein synthesis and inhibiting inflammatory reaction, so as to improve the development of physical burn caused by severe burns, backward learning and memory ability and decreased motor ability.

【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R726.2

【参考文献】