【摘要】：目的通過應(yīng)用280nm LED(Light Emitting Diode,LED發(fā)光二極管)紫外線在大鼠腹部皮瓣產(chǎn)生缺血再灌注損傷后對(duì)大鼠腹部術(shù)區(qū)進(jìn)行局部照射,來觀察280nm LED紫外線對(duì)大鼠皮瓣再灌注損傷后組織的治療效果,并就相應(yīng)的實(shí)驗(yàn)組和對(duì)照組所產(chǎn)生的結(jié)果來探討其相關(guān)及可能的作用機(jī)制。方法將同一批在相同環(huán)境下長成的Wistar健康雄性鼠共40只,隨機(jī)分4組,280nm、320nm的實(shí)驗(yàn)組各1組,空白組1組,10只一組,剩下的10只則可作為同批次相同環(huán)境下成長的替補(bǔ)組大鼠,防止實(shí)驗(yàn)中的大鼠因意外造成皮瓣損傷影響到實(shí)驗(yàn)結(jié)果或死去造成實(shí)驗(yàn)結(jié)果缺失的,為保證實(shí)驗(yàn)的正常進(jìn)行,選擇替補(bǔ)大鼠按照相應(yīng)的實(shí)驗(yàn)方法繼續(xù)完成實(shí)驗(yàn)。手術(shù)及替補(bǔ)組大鼠在實(shí)驗(yàn)正式開始前均養(yǎng)于SPF動(dòng)物實(shí)驗(yàn)室1w,并于術(shù)前1d禁食;10%水合氯醛腹腔麻醉行腹部皮瓣模型制作手術(shù),在所有實(shí)驗(yàn)及空白組大鼠腹部皮膚建立再灌注損傷皮瓣的模型后,對(duì)于280nm和320nm兩個(gè)實(shí)驗(yàn)組大鼠模型建成后即立刻行對(duì)應(yīng)的人工紫外線術(shù)區(qū)皮瓣照射1次,時(shí)間為持續(xù)4 min,后持續(xù)單籠飼養(yǎng),正常飼養(yǎng)進(jìn)食,直至取標(biāo)本處死;從術(shù)后第1天開始每天按時(shí)按量對(duì)兩個(gè)實(shí)驗(yàn)組的腹部皮瓣進(jìn)行相應(yīng)波長的紫外線照射,2次/1d,每次4 min,共8min,持續(xù)7 d;對(duì)于空白組大鼠其皮瓣模型制備成功后,不予任何處理措施,等空白組鼠完全從麻醉中醒后給予單籠正常飼養(yǎng)進(jìn)食;術(shù)后每天進(jìn)行觀察,若出現(xiàn)大鼠啃食皮瓣致皮瓣毀損影響結(jié)果的,及時(shí)按相同實(shí)驗(yàn)方法進(jìn)行相應(yīng)組的實(shí)驗(yàn)進(jìn)行替補(bǔ)大鼠實(shí)驗(yàn);三組實(shí)驗(yàn)鼠都在術(shù)后7 d時(shí)候通過坐標(biāo)貼紙的方法來測算腹部手術(shù)區(qū)域皮瓣的成活面積同時(shí)計(jì)算相應(yīng)皮瓣的成活率;手術(shù)后7d給予各組大鼠處死,并在各組大鼠的腹部相應(yīng)術(shù)區(qū)相應(yīng)的位置,切取皮瓣組織并立即相應(yīng)處理制作成病理切片再行免疫組織化學(xué)染色,后通過顯微鏡及Image-Pro Plus 6.0測算陽性血管內(nèi)皮生長因子(Vascular Endothelial Growth Factor,VEGF)的表達(dá)面積以及所占百分比,使用TUNEL實(shí)驗(yàn)方法檢測組織標(biāo)本中細(xì)胞中的凋亡比率,通過SPSS軟件對(duì)數(shù)據(jù)進(jìn)行處理,獲取結(jié)果。結(jié)果280nm,320nm和空白三組術(shù)后7 d皮瓣的成活率檢測分別為62.38%±8.64%、54.49%±4.48%和39.61%±4.22%,對(duì)應(yīng)的組織中VEGF陽性表數(shù)據(jù)分別為91.39%±9.29%、70.46%±4.02%和34.97%±3.08%,對(duì)應(yīng)分組皮瓣組織中細(xì)胞凋亡率顯示為53.06%±5.64%、64.72%±3.73%和32.82%±1.72%。所有的實(shí)驗(yàn)的對(duì)應(yīng)數(shù)據(jù)指標(biāo)中,280nm、320nm和對(duì)照組相比,P0.05;280nm和320nm兩實(shí)驗(yàn)組比較,P0.05。結(jié)論280nm LED-紫外線局部照射治療大鼠腹部皮瓣缺血再灌注后損傷效果明顯,作用機(jī)制與皮瓣中VEGF表達(dá)量的提高促進(jìn)毛細(xì)血管新生而對(duì)組織的成活有積極作用;經(jīng)紫外線照射的皮瓣組織細(xì)胞凋亡率升高,且紫外線波長較短照射組細(xì)胞凋亡情況較輕,此現(xiàn)象可能與皮膚對(duì)不同波長紫外線的吸收情況不同以及它們自身波長穿透性強(qiáng)弱的特性有關(guān)。
[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% 鹵8.64 鹵54.49% 鹵4.48% and 39.61% 鹵4.22 respectively in 320nm group and 39.61% 鹵4.22% in the blank group. The VEGF positive table data were 91.39% 鹵9.29cius 70.46% 鹵4.02% and 34.97% 鹵3.08 respectively. The cell apoptosis rate in the corresponding group was 53.06% 鹵5.6445% 鹵3.73% and 32.82% 鹵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.
【學(xué)位授予單位】：青島大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R622

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