萝卜硫素对糖尿病小鼠溃疡创面愈合的作用与机制研究  

Therapeutic effect and the underlying molecular mechanism of Sulforaphane on the wound healing in diabetic ulcer mice

作　　者：高文婷 GAO Wenting(Zhejiang Chinese Medical University,Hangzhou 310053,China)

机构地区：[1]浙江中医药大学,浙江杭州310053

出　　处：《浙江实用医学》2023年第4期271-277,292,共8页Zhejiang Practical Medicine

摘　　要：目的探讨萝卜硫素对糖尿病小鼠溃疡创面愈合的影响并研究其作用机制。方法选取48只C57BL/6小鼠,分成正常组、模型组、萝卜硫素组(0.95g/kg)、贝复济组(0.8g/kg),每组再设置7天和14天,共8组,每组各6只。除正常组外,其他组小鼠在高糖高脂饲养下腹腔注射链脲佐菌(STZ)建立糖尿病小鼠模型。所有组别小鼠背部两侧进行直径约8mm圆形皮肤创伤,构建糖尿病溃疡小鼠创面模型。qRT-PCR法检测小鼠创面组织中炎症因子TNF-α、IL-6、IL-10与抗氧化应激因子SOD、CAT、HO-1 mRNA水平;ELISA法检测血清中VEGF、CD31水平;Western Blot法检测创面组织炎症通路蛋白NF-кB、氧化应激通路蛋白Nrf 2表达水平。采用细胞划痕试验检测萝卜硫素对HaCaT细胞迁移的影响;CCK-8法检测萝卜硫素对HaCaT细胞生长的影响。结果与模型组比较,萝卜硫素组和贝复济组糖尿病溃疡小鼠创面愈合速率增加(P<0.05),显著下调TNF-α、IL-6水平(P<0.05),上调IL-10、SOD、CAT、HO-1m RNA水平(P<0.05),增加VEGF、CD31含量(P<0.05);抑制NF-кB表达,促进Nrf 2表达(P<0.05);HaCaT细胞迁移与增殖能力增加(P<0.05)。萝卜硫素组与贝复济组比较,治疗效果相当,差异无统计学意义(P>0.05)。结论萝卜硫素修复糖尿病小鼠溃疡创面的效果显著,可改善溃疡创伤处炎症和氧化应激水平,加快创面愈合。Objective To investigate the effect of Sulforaphane(SFN)on the wound healing of diabetic ulcer in mice and its mechanisms.Methods 48 C57BL/6 mice were selected and divided into four groups:normal group,model group,SFN group(0.95g/kg),and Beifuji group(0.8g/kg),seach group set 7 days group and 14 days group,six mice in each group.Except for the normal group,the mice in the other groups were fed with high sugar and high fat under intraperitoneal injection of streptozotococcus(STZ)to establish a diabetic mice model.All groups of mice underwent approximately 8 mm circular skin wound on both sides of the back to construct the diabetic skin ulcer model.Eyeball blood and wound tissue were collected from each group of mice.qRT-PCR was used to detect the mRNA levels of inflammation-related factors TNF-α、IL-6、IL-10、anti-oxidative stress factors SOD、CAT and HO-1 mRNA;ELISA was used to evaluate the levels of VEGF and CD31 in serum.Western blot was used to detect the expression levels of inflammatory pathway protein NF-кB and oxidative stress pathway protein Nrf 2 in wound tissue.Results Compared with the model group,the administration of SFN and Beifuji significantly accelerated the healing rate of diabetic ulcer mice(P<0.05),down-regulated TNF-α、IL-6 levels(P<0.05)and up-regulated IL-10、SOD、CAT and HO-1 mRNA levels(P<0.05).The expression of VEGF and CD31 is higher in SFN and Beifuji group than model group(P<0.05).The protein level of NF-кB was decreased and the expression of Nrf 2 was decreased after SFN and Beifuji treatment(P<0.05).Also,SFN and Beifuji increased the migration and proliferation of HaCaT cells significantly(P<0.05).Furthermore,the therapeutic effects of Beifuji and SFN in the treatment of diabetic ulcers was comparable,no significant difference were found(P>0.05).Conclusion SFN was effective in repairing diabetic ulcer wounds in mice,reducing the levels of inflammation and oxidative stress at ulcer wounds,accelerating diabetic ulcer mice wound healing.

关 键 词：糖尿病溃疡 创面愈合 萝卜硫素 贝复济 氧化应激 炎症 

分 类 号：R285.5[医药卫生—中药学]