摘要
目的探讨紫色疽疮膏治疗糖尿病足血瘀型慢性疮面的作用机制,观察其对内皮细胞自噬水平的影响。方法选取5例采用紫色疽疮膏治疗的糖尿病足血瘀型慢性疮面患者为研究对象,分别在治疗前及治疗3、6 d后局部蚕食清创处理时留取适量创面组织标本,作为治疗组,另选取肢体近端正常皮肤真皮浅层组织作为正常对照组,通过CD34免疫组织化学染色和免疫荧光四重染色观察比较2组皮肤真皮浅层微血管内径情况,采用蛋白质免疫印迹法观察比较2组创面组织中膜型微管相关蛋白1轻链3(LC3Ⅱ)/胞浆型微管相关蛋白1轻链3(LC3Ⅰ)、内皮型一氧化氮合酶(eNOS)及磷酸化哺乳动物雷帕霉素靶蛋白(p-mTOR)蛋白表达水平情况。结果免疫组织化学染色和免疫荧光四重染色结果均显示,与正常对照组比较,治疗组治疗前皮肤真皮浅层微血管内径明显增大(P<0.05);与治疗组治疗前比较,治疗组治疗3、6 d后皮肤真皮浅层微血管内径均明显减小(P<0.05),且治疗组治疗3 d后与治疗6 d后微血管内径比较差异无统计学意义(P>0.05)。与正常对照组比较,治疗组治疗前创面组织LC3Ⅱ/LC3Ⅰ及p-mTOR蛋白表达水平明显降低(P<0.05),eNOS蛋白表达水平明显升高(P<0.05);与治疗组治疗前比较,治疗组治疗3、6 d后创面组织LC3Ⅱ/LC3Ⅰ及p-mTOR蛋白表达水平明显升高(P<0.05),eNOS蛋白表达水平明显降低(P<0.05),且治疗组治疗3 d后与治疗6 d后各指标比较差异均无统计学意义(P>0.05)。结论紫色疽疮膏治疗糖尿病足血瘀型慢性创面疗效确切,其作用机制可能与增加LC3Ⅱ/LC3Ⅰ及p-mTOR蛋白表达水平,降低eNOS蛋白表达水平,提高内皮细胞自噬水平有关,可改善血管内皮细胞的舒缩功能,改善创面局部血管扩张淤滞状态,改变局部微环境,促进血液循环,促进新陈代谢,进而促进创面的生长和修复。
Objective To study the mechanism of Purple Juchuang Ointment on chronic wounds of diabetic foot with blood stasis type,and explore its effect on the autophagy level of endothelial cells.Methods Totally 5 diabetic foot patients with blood stasis type,who were used Purple Juchuang Ointment to treat chronic sores,were selected as the research subjects.When debridement was performed on the local encroachment of their diabetic foot,appropriate tissue specimens at each time point(before treatment,3 and 6 days after treatment)were retained,and the collected tissue specimens were treated as the treatment group;in addition,the normal superficial dermal tissue of the proximal skin of the limb in diabetic foot of these 5 patients was treated as the normal control group.CD34 immunohistochemical staining and immunofluorescence quadruple staining were used to observe and compare the internal diameter of superficial dermal microvessels between the two groups.Western blotting was used to observe and compare the levels of membrane microtubule-associated protein 1 light chain 3(LC3Ⅱ),cytoplasmic microtubule-associated protein 1 light chain 3(LC3Ⅰ),endothelial nitric oxide synthase(eNOS)and phosphorylated mammalian target of rapamycin(p-mTOR).Results The results immunohisto chemical staining and immunofluorescence quadruple staining all showed that:compared with the normal control group,the inner diameters of superficial dermal microvessels in the treatment group were significantly increased before treatment(P<0.05).Compared with before treatment,the inner diameters of superficial dermal microvessels in the treatment group were significantly reduced after 3 and 6 days of treatment(P<0.05),with no statistical difference(P>0.05).Compared with the normal control group,the level of LC3Ⅱ/LC3Ⅰand p-mTOR in the wound tissue in the treatment group before treatment was significantly reduced(P<0.05),and the level of eNOS was significantly increased(P<0.05).Compared with the treatment group before treatment,the levels of LC3Ⅱ/LC3Ⅰ
作者
王广宇
王雨
王雷永
徐旭英
WANG Guangyu;WANG Yu;WANG Leiyong;XU Xuying(Department of Ulcer Vascular surgery,Beijing Hospital of Traditional Chinese Medicine,Capital Medical University,Beijing 100010)
出处
《河北中医》
2021年第1期56-60,68,共6页
Hebei Journal of Traditional Chinese Medicine
基金
国家自然科学基金委员会资助项目(面上项目)(编号:81673975)。
关键词
糖尿病足
皮肤溃疡
自噬
中药疗法
Diabetic foot
Skin ulcer
Autophagy
Traditional Chinese medicin therapy
