葉酸對(duì)糖尿病小鼠皮膚創(chuàng)面愈合延遲的保護(hù)作用
發(fā)布時(shí)間：2018-07-23 13:30
【摘要】：目的采用鏈脲佐菌素（STZ）建立糖尿病小鼠模型，并在此基礎(chǔ)上構(gòu)建糖尿病小鼠皮膚創(chuàng)面愈合延遲模型，探討補(bǔ)充葉酸對(duì)糖尿病小鼠皮膚創(chuàng)面愈合延遲的效應(yīng)并初步探討其機(jī)制。 方法ICR小鼠分為正常對(duì)照小鼠及建糖尿病模型小鼠，建糖尿病模型小鼠連續(xù)5日腹腔注射STZ50mg/kg，正常對(duì)照小鼠連續(xù)5日腹腔注射檸檬酸緩沖液。首次注射前及末次注射后72h采集小鼠尾靜脈血液檢測(cè)血糖濃度，以血糖濃度高于250mg/dl判定為糖尿病模型構(gòu)建成功。在此基礎(chǔ)上，正常對(duì)照小鼠納入正常對(duì)照組（Control組），建糖尿病模型小鼠分為2組：糖尿病對(duì)照組（DM組）、糖尿病葉酸處理組（DM+FA組），，所有小鼠均制作皮膚創(chuàng)面，以創(chuàng)面制作成功之日定為第0天，實(shí)驗(yàn)過程中每日觀察小鼠皮膚創(chuàng)面愈合情況，包括創(chuàng)面濕潤(rùn)度、顏色、分泌物、形狀、創(chuàng)緣；于第0、3、6、9、12天以透明薄膜準(zhǔn)確描繪創(chuàng)面，計(jì)算各時(shí)間點(diǎn)創(chuàng)面愈合率；于第0、3、6、9、12天，每組各6只小鼠麻醉后處死，取相同部位創(chuàng)面組織用于組織學(xué)檢查，第3、6、9、12天部分創(chuàng)面組織用于測(cè)定羥脯氨酸含量；于第6天每組各6只小鼠麻醉后處死，留取創(chuàng)面組織測(cè)定谷胱甘肽（GSH）、丙二醛（MDA）、一氧化氮（NO）含量及采用Western blotting測(cè)定3-硝基酪氨酸（3-NT）蛋白表達(dá)水平，留取肝臟組織測(cè)定GSH含量；每組剩余16只小鼠持續(xù)觀察至創(chuàng)面完全愈合，記錄創(chuàng)面完全愈合時(shí)間，并于實(shí)驗(yàn)結(jié)束前再次采集小鼠尾靜脈血液，檢測(cè)血糖濃度。 結(jié)果建糖尿病模型小鼠連續(xù)5日腹腔注射STZ后小鼠精神狀態(tài)變差，毛發(fā)失去光澤，枯黃松散，體重減輕，末次注射后72h血糖濃度高于250mg/dl，實(shí)驗(yàn)結(jié)束前再次檢測(cè)小鼠血糖濃度仍高于250mg/dl。在糖尿病小鼠模型建立成功基礎(chǔ)上制作皮膚創(chuàng)面，Control組小鼠皮膚創(chuàng)面完全愈合時(shí)間為（12.08±1.08）天，DM組為（14.00±2.00）天與Control組比較明顯延遲，DM+FA組創(chuàng)面完全愈合時(shí)間為（12.73±0.79）天與DM組比較明顯縮短；第3、6、9、12天各時(shí)間點(diǎn)創(chuàng)面愈合率比較顯示Control組與DM+FA組兩組之間無差異，Control組與DM組、DM+FA組與DM組之間比較各時(shí)間點(diǎn)差異均有統(tǒng)計(jì)學(xué)意義；形態(tài)學(xué)及組織學(xué)觀察可見DM組小鼠皮膚創(chuàng)面愈合延遲，補(bǔ)充葉酸改善創(chuàng)面愈合；第3、6、9、12天三組小鼠創(chuàng)面組織羥脯氨酸含量均呈遞增趨勢(shì)，DM組與Control組比較各時(shí)間點(diǎn)羥脯氨酸含量均顯著降低，DM+FA組與DM組比較第6、9、12天羥脯氨酸含量升高。測(cè)定第6天三組小鼠創(chuàng)面皮膚組織GSH、MDA、NO含量和3-NT蛋白表達(dá)水平結(jié)果顯示，與Control組相比較，DM組MDA含量升高、GSH含量降低、NO含量降低、3-NT蛋白表達(dá)水平升高，補(bǔ)充葉酸后可降低小鼠創(chuàng)面皮膚組織MDA、3-NT水平，NO水平升高，降低機(jī)體GSH耗竭，減輕機(jī)體氧化損傷。 結(jié)論本研究得出以下結(jié)論：（1）補(bǔ)充葉酸對(duì)糖尿病小鼠皮膚創(chuàng)面愈合延遲具有保護(hù)作用；（2）葉酸對(duì)糖尿病小鼠愈合延遲的保護(hù)作用機(jī)制可能與葉酸減輕創(chuàng)面局部氧化應(yīng)激，提高創(chuàng)面組織NO水平有關(guān)。
[Abstract]:Objective to establish a diabetic mouse model with streptozotocin (STZ), and to establish a delayed skin wound healing model in diabetic mice, and to explore the effect of folic acid supplementation on the delayed healing of diabetic skin wound and its mechanism. Methods ICR mice were divided into normal control mice and diabetic model mice. STZ 50 mg / kg was injected intraperitoneally for 5 days and citric acid buffer solution was injected intraperitoneally for 5 days in normal control mice. The blood glucose concentration of tail vein of mice was detected before the first injection and 72 hours after the last injection. The blood glucose concentration was higher than that of 250mg/dl and the diabetic model was successfully constructed. On this basis, the normal control mice were included in the normal control group (Control group), and the diabetic model mice were divided into two groups: diabetic control group (DM group) and diabetic folic acid treated group (DM FA group). The wound healing was observed daily in mice, including wound wetness, color, secretion, shape and wound margin. The wound healing rate was calculated at each time point, at the 12th day after anesthesia, 6 mice in each group were killed, the wound tissue of the same site was taken for histological examination, and the content of hydroxyproline was measured in some wound tissues on the 12th day of the 3rd day. Six mice in each group were killed after anesthesia on the 6th day. The content of glutathione (GSH), malondialdehyde (MDA),) nitric oxide (MDA),) (NO) and the expression of 3-nitrotyrosine (3-NT) protein were measured by Western blotting and GSH content in liver tissue was determined by Western blotting. The remaining 16 mice in each group were observed to heal completely and the time of complete wound healing was recorded. The blood samples of caudal vein of mice were collected again before the end of the experiment to detect the concentration of blood glucose. Results after intraperitoneal injection of STZ for 5 consecutive days, the diabetic model mice showed poor mental state, lost luster of hair, loose yellow and weight loss. The concentration of blood glucose was higher than 250 mg / dl at 72 h after the last injection, and the concentration of blood sugar in mice was still higher than 250 mg / dl before the end of the experiment. The complete healing time of skin wound in control group was (12.08 鹵1.08) days and that in DM group was (14.00 鹵2.00) days, which was significantly shorter than that in Control group (12.73 鹵0.79) days. The comparison of wound healing rate between Control group and DM FA group showed that there was no difference between Control group and DM FA group. Morphological and histological observation showed that skin wound healing was delayed and folic acid supplementation improved wound healing in DM group. The content of hydroxyproline in wound tissue of DM group and Control group was significantly lower than that of DM FA group and DM group on the 12th day, and the hydroxyproline content in DM FA group was significantly higher than that in DM group. On the 6th day, the levels of GSH MDAN no and 3-NT protein expression in the skin tissue of the three groups were measured. The results showed that compared with the Control group, the MDA content in DM group was higher than that in the DM group, and the no content in DM group was lower than that in the DM group and the protein expression level of 3-NT was decreased. After folic acid supplementation, the level of MDA-3-NT in skin tissue of mice was decreased, the level of no increased, the depletion of GSH was decreased, and the oxidative damage of the body was alleviated. Conclusion the conclusions are as follows: (1) folic acid supplementation has protective effect on delayed wound healing in diabetic mice; (2) the protective mechanism of folic acid on delayed healing of diabetic mice may be associated with folic acid in relieving local oxidative stress. It is related to the increase of no level in wound tissue.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R622;R587.1
[Abstract]:Objective to establish a diabetic mouse model with streptozotocin (STZ), and to establish a delayed skin wound healing model in diabetic mice, and to explore the effect of folic acid supplementation on the delayed healing of diabetic skin wound and its mechanism. Methods ICR mice were divided into normal control mice and diabetic model mice. STZ 50 mg / kg was injected intraperitoneally for 5 days and citric acid buffer solution was injected intraperitoneally for 5 days in normal control mice. The blood glucose concentration of tail vein of mice was detected before the first injection and 72 hours after the last injection. The blood glucose concentration was higher than that of 250mg/dl and the diabetic model was successfully constructed. On this basis, the normal control mice were included in the normal control group (Control group), and the diabetic model mice were divided into two groups: diabetic control group (DM group) and diabetic folic acid treated group (DM FA group). The wound healing was observed daily in mice, including wound wetness, color, secretion, shape and wound margin. The wound healing rate was calculated at each time point, at the 12th day after anesthesia, 6 mice in each group were killed, the wound tissue of the same site was taken for histological examination, and the content of hydroxyproline was measured in some wound tissues on the 12th day of the 3rd day. Six mice in each group were killed after anesthesia on the 6th day. The content of glutathione (GSH), malondialdehyde (MDA),) nitric oxide (MDA),) (NO) and the expression of 3-nitrotyrosine (3-NT) protein were measured by Western blotting and GSH content in liver tissue was determined by Western blotting. The remaining 16 mice in each group were observed to heal completely and the time of complete wound healing was recorded. The blood samples of caudal vein of mice were collected again before the end of the experiment to detect the concentration of blood glucose. Results after intraperitoneal injection of STZ for 5 consecutive days, the diabetic model mice showed poor mental state, lost luster of hair, loose yellow and weight loss. The concentration of blood glucose was higher than 250 mg / dl at 72 h after the last injection, and the concentration of blood sugar in mice was still higher than 250 mg / dl before the end of the experiment. The complete healing time of skin wound in control group was (12.08 鹵1.08) days and that in DM group was (14.00 鹵2.00) days, which was significantly shorter than that in Control group (12.73 鹵0.79) days. The comparison of wound healing rate between Control group and DM FA group showed that there was no difference between Control group and DM FA group. Morphological and histological observation showed that skin wound healing was delayed and folic acid supplementation improved wound healing in DM group. The content of hydroxyproline in wound tissue of DM group and Control group was significantly lower than that of DM FA group and DM group on the 12th day, and the hydroxyproline content in DM FA group was significantly higher than that in DM group. On the 6th day, the levels of GSH MDAN no and 3-NT protein expression in the skin tissue of the three groups were measured. The results showed that compared with the Control group, the MDA content in DM group was higher than that in the DM group, and the no content in DM group was lower than that in the DM group and the protein expression level of 3-NT was decreased. After folic acid supplementation, the level of MDA-3-NT in skin tissue of mice was decreased, the level of no increased, the depletion of GSH was decreased, and the oxidative damage of the body was alleviated. Conclusion the conclusions are as follows: (1) folic acid supplementation has protective effect on delayed wound healing in diabetic mice; (2) the protective mechanism of folic acid on delayed healing of diabetic mice may be associated with folic acid in relieving local oxidative stress. It is related to the increase of no level in wound tissue.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R622;R587.1
【共引文獻(xiàn)】
相關(guān)期刊論文 前10條
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