化腐再生法调控iNOS、IL-1β、CD16的表达促进糖尿病大鼠筋之血化  

Effect of Huafu Zaisheng Method on Tendon Vascularization by Regulating iNOS,IL-1β,CD16 Expression in Diabetic Rats

作　　者：孙旭[1] 孙瀚驰 徐强[2] 朱朝军[2] 刘潇鸽 宋子豪 李萌[1] 刘振雷 张朝晖[2] SUN Xu;SUN Hanchi;XU Qiang;ZHU Chaojun;LIU Xiaoge;SONG Zihao;LI Meng;LIU Zhenlei;ZHANG Zhaohui(Tianjin University of Traditional Chinese Medicine,Tianjin China 301617;The Second Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin China 300250)

机构地区：[1]天津中医药大学,天津301617 [2]天津中医药大学第二附属医院,天津300250

出　　处：《中医学报》2021年第10期2166-2172,共7页Acta Chinese Medicine

基　　金：国家重点研发计划项目(2019YFC1709303);国家自然科学基金面上项目(81573972);天津中医药大学第二附属医院“青苗计划”项目(QMJH202001);河北省中医药管理局2022年度中医药类科研计划项目(2022554)。

摘　　要：目的:观察化腐再生法调控糖尿病大鼠溃疡局部创面促筋之血化过程中诱导型一氧化氮合成酶(inducible nitric ox-ide synthase,iNOS)、白细胞介素-1β(interleukin-1β,IL-1β)、CD16的表达。方法:30只SD大鼠随机分为模型组、阳性对照组、治疗1组、治疗2组、治疗3组,每组6只,所有大鼠腹腔注射链脲佐菌素(streptozotocin,STZ)建立糖尿病模型后,采用外科方法在大鼠腰背部脊柱两侧制作2个伴肌腱损伤的溃疡创面并滴加50μL冰醋酸到创面上造成糖尿病大鼠肌腱变性坏死溃疡模型。模型组采用凡士林油纱外敷,阳性对照组给予水凝胶外涂,治疗1组外涂橡皮生肌膏,治疗2组外敷菠萝蛋白酶撒布,治疗3组橡皮生肌膏外涂联合菠萝蛋白酶撒布外敷,每日1次,连续21 d。分别于治疗后第3 d、7 d、14 d、21 d,眼眶采血,每次1.5 mL,ELISA法检测血清中iNOS、IL-1β、CD16的水平;免疫组化法检测不同时期大鼠创周组织中iNOS、IL-1β、CD16的蛋白表达水平。结果:除模型组外,其余大鼠血清中iNOS、IL-1β、CD16的水平均呈现先上升后下降的趋势,其中,治疗第3 d时,治疗3组大鼠血清中iNOS、IL-1β、CD16的水平显著高于模型组(P<0.05);治疗21 d时,治疗3组大鼠血清中iNOS、IL-1β、CD16的水平显著低于模型组(P<0.05)。治疗3 d时,大鼠创周组织iNOS、IL-1β、CD16的蛋白表达水平显著高于模型组(P<0.05),且随着治疗周期的延长,蛋白表达水平呈先升高后降低的趋势。结论:化腐再生法具有早期促使iNOS、IL-1β、CD16释放,而后期抑制其表达的双向调节作用,从而有效调控糖尿病大鼠筋之血化过程。Objective:To observe the effect of Huafu Zaisheng method on tendon′s vascularization of local ulcer wounds in diabetic rats by inducing the expression of inducible nitric oxide synthase(iNOS),interleukin 1β(IL-1β),and CD 16.Methods:30 SD rats were randomly divided into positive control group,model group,treatment group 1,treatment group 2,and treatment group 3,with 6 rats in each.After all the rats were intraperitoneally injected with streptozotocin(STZ)to establish a diabetes model,50μL of glacial acetic acid was added to the wound surface by surgical method to form the tendon degeneration and necrosis ulcer model of diabetic rat.The positive control group was dressing changed with hydrogel,the model group with Vaseline gauze,treatment group 1 with skin-producing adhesive plaster by external application,the treatment group 2 with bromelain scattering,and the treatment group 3 was dressing changed with external application of skin-producing adhesive plaster combined with bromelain scattering,once a day.Then on the 3 rd,7 th,14 th,and 21 st days after treatment,blood was collected from the rat′s orbit,1.5 mL each time.The levels of iNOS,IL-1β,and CD16 in the serum were detected by ELISA.And the peri-wound tissues of the rats were extracted,and immunohistochemical methods were used to detect the expression of iNOS,IL-1β,and CD16 in different periods.Results:Except for the model group,the serum levels of iNOS,IL-1β,and CD16 in the rest rats were increased first and then decreased.Among them,the serum levels of iNOS,IL-1β,and CD16 in the rats of the treatment 3 group were significantly higher than those in the model group on the 3 rd,7 th,and 14 th days of treatment(P<0.05).On the 21 st day of treatment,the serum levels of iNOS,IL-1β,and CD16 in group 3 were significantly higher than that in the model group(P<0.05).And the immunohistochemistry results showed that the protein expression levels of iNOS,IL-1β,and CD16 in the peri-wound tissue of rats were significantly higher than the model group(P<0.05).Co

关 键 词：化腐再生法 糖尿病溃疡 筋之血化 双向调节 橡皮生肌膏 菠萝蛋白酶 大鼠 

分 类 号：R285.5[医药卫生—中药学]