摘要
【目的】观察化腐再生理论指导下菠萝蛋白酶与橡皮生肌膏对糖尿病大鼠伴肌腱坏死溃疡创面出现筋之血化过程中局部皮肤与肌腱组织血管内皮生长因子(VEGF)表达的影响,探讨化腐再生法调控局部微环境构建血管新生促进"筋之血化"形成的机制。【方法】将30只大鼠随机分为对照组、模型组、再生组、化腐组、化腐再生组,每组6只。各组均采用链脲佐菌素(STZ)诱导构建糖尿病大鼠模型,再采用外科方法建立糖尿病大鼠肌腱坏死溃疡模型。造模结束后,以上各组分别对应给予水凝胶、凡士林油纱、橡皮生肌膏、菠萝蛋白酶、菠萝蛋白酶+橡皮生肌膏外敷治疗,共21 d。计算创面愈合率,记录出现筋之血化及创面愈合的时间,分别于治疗第3、7、14、21天,采用免疫组织化学法检测创面皮肤与肌腱组织VEGF阳性表达以及平均光密度值(AOD),苏木素-伊红(HE)染色法观察创面皮肤与肌腱组织病理变化。【结果】化腐再生组创面愈合率最高,与其余4组比较,差异均有统计学意义(P<0.05)。化腐再生组出现筋之血化时间明显短于模型组和化腐组,治疗第7、14天肌腱组织VEGF阳性表达强于模型组,AOD值高于模型组(均P<0.05),HE染色结果可见少量肉芽组织,肌腱附近可见较多新生小血管,有少量炎性细胞浸润,化腐组、再生组未见新生小血管。化腐再生组创面愈合时间最短,与其余4组比较,差异均有统计学意义(P<0.05)。治疗第3、14、21天,各组创面皮肤组织VEGF的AOD值组间差异均无统计学意义(P>0.05)。治疗第7天,化腐再生组创面皮肤组织VEGF阳性表达强于模型组,AOD值高于模型组(P<0.05),HE染色结果可见较多新生小血管;治疗第14天,化腐再生组可见大量肉芽组织,对照组、化腐组、再生组可见较多新生小血管;治疗第21天,除模型组外,其他各组表皮层结构较完整,可见毛囊、皮脂腺等附属器官,未见明显炎症。【结论】化腐再生法可促进糖尿病大鼠伴肌腱坏死溃疡创面出现筋之血化及创面愈合,其机制可能与其在创面愈合早中期(7~14 d)促进VEGF表达有关。应用菠萝蛋白酶与橡皮生肌膏治疗糖尿病大鼠伴肌腱坏死溃疡创面,为筋之血化的出现及创面愈合提供了有利条件。
Objective To observe the effect of bromelain and Xiangpi Shengji Ointment on the expression of vascular endothelial growth factor(VEGF)in local skin and tendon tissues during the vascularization of tendon produced in wound surface of diabetes mellitus rats complicated with tendon necrosis ulcer under the guidance of putridity-resolving and regeneration theory,so as to explore the mechanism of putridity-resolving and regenerative therapyregulatinglocalmicroenvironmenttoconstructangiogenesisthuspromotingtheformationofvascularization of tendon. Methods Thirty SD rats were randomly divided into control group, model group, regeneration group,putridity-resolving group,and putridity-resolving and regeneration group,6 rats in each group. All the rats in the above groups were induced into diabetes mellitus model by intraperitoneal injection of streptozotocin(STZ),and then further induced into diabetes mellitus model complicated with tendon necrosis ulcer. After modeling, the control group, model group, regeneration group, putridity-resolving group, and putridity-resolving and regeneration group were given external use of hydrogel,vaseline gauze,Xiangpi Shengji Ointment,bromelain,and bromelain combined with Xiangpi Shengji Ointment,respectively,for 21 days. The wound healing rate was calculated,and the time for vascularization of tendon and wound healing were recorded. On treatment day 3,7,14,and 21,the positive expression level and average optical density(AOD)of VEGF in the ulcer skin and tendon tissues were determined by immunohistochemistry,and the pathological features of ulcer skin and tendon tissues were observed by haematoxylin-eosin(HE)staining. Results The wound healing rate in the putridity-resolving and regeneration group was the highest,and the difference was statistically significant as compared with that in the other 4 groups(P<0.05). The time for vascularization of tendon in the putridity-resolving and regeneration group was significantly shorter than that in the model group and putridity-resolving group,positive expression degree and AOD value of VEGF in tendon tissue on treatment day 7 and 14 were higher than those in the model group on the same time(P<0.05),and the HE results showed that a small amount of granulation tissue,more new small blood vessels and a small number of inflammatory cells infiltration near the tendon were seen. No new small blood vessels were found in the putridity-resolving group and regeneration group. The wound healing time in the putridityresolving and regeneration group was the shortest,and the difference was statistically significant as compared with that in the other 4 groups(P<0.05). On treatment day 3,14,21,there was no significant difference in AOD value of VEGF among the various groups(P>0.05). On treatment day 7,the positive expression degree and AOD value of VEGF in the skin tissue of the putridity-resolving and regeneration group were higher than those of the model group(P<0.05), and HE staining results showed that more new small blood vessels were seen. On treatment day 14,a large amount of granulation tissue was seen in the putridity-resolving and regeneration group,and many new small blood vessels were seen in the control group, putridity-resolving group and regeneration group. On treatment day 21, apart from the model group, the relatively complete epidermal structure, and accessory organs involving hair follicles and sebaceous glands in the other 4 groups were found,and no obvious inflammation was seen. Conclusion Putridity-resolving and regeneration therapy has effects on promoting the vascularization of tendon and wound healing in diabetes mellitus rat model complicated with tendon necrosis ulcer,and its mechanism may be related to promoting the expression of VEGF in the early and middle stage(7-14 days)of wound healing. The application of bromelain and Xiangpi Shengji Ointment for the treatment of diabetes mellitus rat model complicated with tendon necrosis ulcer provides favorable conditions for the vascularization of tendon and wound healing.
作者
宋子豪
孙旭
徐强
孙瀚驰
朱朝军
张朝晖
SONG Zi-Hao;SUN Xu;XU Qiang;SUN Han-Chi;ZHU Chao-Jun;ZHANG Zhao-Hui(Postgraduate School of Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;The Second Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300250,China)
出处
《广州中医药大学学报》
CAS
2020年第9期1737-1746,共10页
Journal of Guangzhou University of Traditional Chinese Medicine
基金
国家自然科学基金面上项目(编号:81573972)。
关键词
化腐再生
筋之血化
糖尿病
溃疡
肌腱坏死
菠萝蛋白酶
橡皮生肌膏
血管内皮生长因子
大鼠
putridity-resolving and regeneration
vascularization of tendon
diabetes mellitus
ulcer
tendon necroesis
bromelain
Xiangpi Shengji Ointment
vascular endothelial growth factor(VEGF)
rats
