龙血竭促进糖尿病溃疡大鼠创面修复的机制研究

Mechanism of dragon blood on promoting the repair of diabetic ulcer wound rats

目的研究龙血竭对糖尿病溃疡创面的修复作用,及其对细胞增殖与凋亡的影响。方法将60只清洁级Wistar雄性大鼠随机分为正常组、模型组和实验组,每组20只。适应性饲养1周后,在大鼠背部后端正中皮下植入圆片形磁铁(直径13 mm、厚2 mm),其中模型组和实验组连续3 d腹腔注射30 mg·kg-1链脲霉素(STZ)诱导糖尿病,1周后测量随机血糖大于16. 7 mmol·L-1的大鼠为糖尿病大鼠。每只大鼠在植入磁片对应的皮肤外直接添加外源磁片,进行溃疡诱导实验,以皮肤变黑、变硬、针刺不出血作为溃疡形成的标准。正常组和模型组创面每天予凡士林纱布换药,实验组予龙血竭胶囊内粉末外用于创面,每天换药1次。在实验开始后的第3,5,7,10,14天计算创面愈合率,进行HE染色观察创面组织病理形态学,第7,14天用SABC三步法检测创面组织中B细胞淋巴瘤-2蛋白(Bcl-2)、Bcl-2相关X蛋白(Bax)、转化生长因子-β1蛋白(TGF-β1)及α-平滑肌肌动蛋白(α-SMA)表达水平。结果实验开始后第14天,正常组、模型组和实验组的创面愈合率分别为(93. 03±4. 37)%,(56. 39±3. 71)%和(85. 93±4. 92)%,实验组与模型组比较差异有统计学意义(P <0. 05)。实验第14天正常组胶原纤维数量增加最明显,实验组次之;正常组与实验组表面上皮完全愈合,模型组溃疡愈合不佳。实验第7天,正常组与实验组Bcl-2表达明显,与模型组相比差异有统计学意义(P <0. 05),而模型组Bax表达最强,与正常组和实验组比较差异有统计学意义(P <0. 05),表明实验组能上调Bcl-2表达、下调Bax表达。实验第7天,正常组与实验组TGF-β1与α-SMA表达水平显著高于模型组,差异有统计学意义(P <0. 05)。结论龙血竭能有效促进糖尿病溃疡创面修复。

Objective To explore the effect of dragon blood on the repair of diabetic ulcer wound and its effect on cell proliferation and apoptosis.Methods Sixty Wistar male rats of clean grade were randomly divided into three groups:normal group,model group and experimental group.One week after adaptive feeding,a disc-shaped magnet(diameter:13 mm,thickness:2 mm)was implanted under the median skin of the rat back.After wound healing,model group and experimental group were injected intraperitoneally with 30 mg·kg-1 streptozotocin(STZ)continued for 3 d to induce diabetes.After one week,the rat was regarded as a diabetic rat,whose blood glucose was measured greater than 16.7 mmol·L-1.Each rat was directly added with an exogenous magnetic piece outside the skin corresponding to the magnetic disk,and an ulcer induction experiment was performed.The skin became dark,hardened,and would not bleed by acupuncture as a standard for ulcer formation.The normal group and model group were given vaseline gauze dressing once a day.The experimental group was given the inside of the dragon blood capsule for the wound,and the dressing was changed once a day.The wound healing rate was calculated on the 3,5,7,10 and 14 day after the start of the experiment,and the pathological morphology of the wound was observed by HE stain.The expression levels of B-cell lymphoma-2 protein(Bcl-2),Bcl-2 associated X protein(Bax),transforming growth factor-β1 protein(TGF-β1)andα-smooth muscle actin protein(α-SMA)in wound tissue were detected by SABC three-step method on 7 and 14 day.Results The wound healing rates of normal group,model group and experimental group on 14 day after experiment were(93.03±4.37)%,(56.39±3.71)%and(85.93±4.92)%.There was significant difference between experimental group and model group(P<0.05).On 14 day of experiment,the increase of the number of collagen fibers in the normal group was the most obvious,followed by the experimental group.The surface epithelium of the normal group and the experimental group completely healed,and the ulcer in the model group healed poorly.On the 7 day of experiment,the expression of Bcl-2 in normal group and experimental group were significant stronger than model group(P<0.05),the expression of Bax in model group was the strongest,and the difference between experimental group and model group was statistically significant(P<0.05).On 7 day of the experiment,the expression of TGF-β1 andα-SMA in normal group and experimental group were stronger than model group,and the difference was statistically significant(P<0.05).Conclusion Dragon blood can effectively promote the repair of diabetic ulcer wounds.