280nmLED-紫外线应用于皮瓣缺血再灌注损伤后治疗的实验研究

发布时间：2018-09-06 13:00

【摘要】：目的通过应用280nm LED(Light Emitting Diode,LED发光二极管)紫外线在大鼠腹部皮瓣产生缺血再灌注损伤后对大鼠腹部术区进行局部照射,来观察280nm LED紫外线对大鼠皮瓣再灌注损伤后组织的治疗效果,并就相应的实验组和对照组所产生的结果来探讨其相关及可能的作用机制。方法将同一批在相同环境下长成的Wistar健康雄性鼠共40只,随机分4组,280nm、320nm的实验组各1组,空白组1组,10只一组,剩下的10只则可作为同批次相同环境下成长的替补组大鼠,防止实验中的大鼠因意外造成皮瓣损伤影响到实验结果或死去造成实验结果缺失的,为保证实验的正常进行,选择替补大鼠按照相应的实验方法继续完成实验。手术及替补组大鼠在实验正式开始前均养于SPF动物实验室1w,并于术前1d禁食;10%水合氯醛腹腔麻醉行腹部皮瓣模型制作手术,在所有实验及空白组大鼠腹部皮肤建立再灌注损伤皮瓣的模型后,对于280nm和320nm两个实验组大鼠模型建成后即立刻行对应的人工紫外线术区皮瓣照射1次,时间为持续4 min,后持续单笼饲养,正常饲养进食,直至取标本处死;从术后第1天开始每天按时按量对两个实验组的腹部皮瓣进行相应波长的紫外线照射,2次/1d,每次4 min,共8min,持续7 d;对于空白组大鼠其皮瓣模型制备成功后,不予任何处理措施,等空白组鼠完全从麻醉中醒后给予单笼正常饲养进食;术后每天进行观察,若出现大鼠啃食皮瓣致皮瓣毁损影响结果的,及时按相同实验方法进行相应组的实验进行替补大鼠实验;三组实验鼠都在术后7 d时候通过坐标贴纸的方法来测算腹部手术区域皮瓣的成活面积同时计算相应皮瓣的成活率;手术后7d给予各组大鼠处死,并在各组大鼠的腹部相应术区相应的位置,切取皮瓣组织并立即相应处理制作成病理切片再行免疫组织化学染色,后通过显微镜及Image-Pro Plus 6.0测算阳性血管内皮生长因子(Vascular Endothelial Growth Factor,VEGF)的表达面积以及所占百分比,使用TUNEL实验方法检测组织标本中细胞中的凋亡比率,通过SPSS软件对数据进行处理,获取结果。结果280nm,320nm和空白三组术后7 d皮瓣的成活率检测分别为62.38%±8.64%、54.49%±4.48%和39.61%±4.22%,对应的组织中VEGF阳性表数据分别为91.39%±9.29%、70.46%±4.02%和34.97%±3.08%,对应分组皮瓣组织中细胞凋亡率显示为53.06%±5.64%、64.72%±3.73%和32.82%±1.72%。所有的实验的对应数据指标中,280nm、320nm和对照组相比,P0.05;280nm和320nm两实验组比较,P0.05。结论280nm LED-紫外线局部照射治疗大鼠腹部皮瓣缺血再灌注后损伤效果明显,作用机制与皮瓣中VEGF表达量的提高促进毛细血管新生而对组织的成活有积极作用;经紫外线照射的皮瓣组织细胞凋亡率升高,且紫外线波长较短照射组细胞凋亡情况较轻,此现象可能与皮肤对不同波长紫外线的吸收情况不同以及它们自身波长穿透性强弱的特性有关。
[Abstract]:Objective to observe the therapeutic effect of 280nm LED (Light Emitting Diode,LED ultraviolet radiation (UV) on the tissue of rat skin flap after ischemia reperfusion injury. The correlation and possible mechanism of the results of the experimental and control groups were discussed. Methods A total of 40 healthy Wistar male rats were randomly divided into 4 groups (n = 4) with 280 nm in each group. The blank group (n = 10) and the control group (n = 10) were randomly divided into two groups: the control group (n = 10) and the control group (n = 10). In order to ensure the normal conduct of the experiment, the replacement rats were selected to continue the experiment according to the corresponding experimental method. The rats in the operation and replacement groups were kept in the SPF animal laboratory for 1 week before the beginning of the experiment. The abdominal flap model was made by celiac anaesthesia with 10% chloral hydrate on the 1st day before operation. After all the experiments and the blank group established the model of reperfusion injury skin flap of the abdominal skin of the rats, the two experimental groups 280nm and 320nm were irradiated with the corresponding artificial ultraviolet skin flap once immediately after the establishment of the model. The time was 4 min, after feeding in a single cage and feeding normally until the specimens were killed. From the first day after operation, the abdominal flaps of the two experimental groups were irradiated with ultraviolet rays of the corresponding wavelength for 2 times / 1 day, each for 4 min, for 8 minutes, lasting 7 days. After the establishment of the model of the skin flap in the blank group, no treatment measures should be taken. The rats in the blank group were given normal feeding and feeding with a single cage after waking up completely from anesthesia, and observed every day after operation, if there were the results of skin flap damage caused by gnawing the skin flap in rats, Three groups of experimental rats were used to calculate the survival area of the abdominal region flap and the survival rate of the corresponding flap was calculated 7 days after operation by the method of coordinate sticker. On the 7th day after operation, the rats in each group were sacrificed, and the skin flap tissue was cut and processed into pathological sections at the corresponding location of the abdominal area of the rats in each group, and then immunohistochemical staining was performed. The area and percentage of positive vascular endothelial growth factor (Vascular Endothelial Growth Factor,VEGF) expression were measured by microscope and Image-Pro Plus 6.0. The apoptotic ratio in tissue samples was detected by TUNEL method. The data were processed by SPSS software. Get the result. Results the survival rate of the skin flap was 62.38% 卤8.64 卤54.49% 卤4.48% and 39.61% 卤4.22 respectively in 320nm group and 39.61% 卤4.22% in the blank group. The VEGF positive table data were 91.39% 卤9.29cius 70.46% 卤4.02% and 34.97% 卤3.08 respectively. The cell apoptosis rate in the corresponding group was 53.06% 卤5.6445% 卤3.73% and 32.82% 卤1.722% respectively. The corresponding data of all the experiments were compared with that of the control group (P 0.05, 280 nm) and 320nm group (P 0.05). Conclusion the effect of 280nm LED- ultraviolet irradiation on abdominal flap injury after ischemia reperfusion is obvious. The mechanism and the increase of VEGF expression in the flap can promote capillary neovascularization and play a positive role in tissue survival. The apoptosis rate of the skin flap irradiated by ultraviolet ray was increased, and the apoptosis rate of the skin flap irradiated by ultraviolet radiation was lighter than that of the group with shorter ultraviolet wave length. This phenomenon may be related to the different absorption of ultraviolet rays at different wavelengths and their own characteristics of wavelength penetration.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R622

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