局部应用PTD-SOD对机械性创伤愈合的影响

目的 探讨氧化应激在机械性创伤愈合发展中的地位、作用和抗氧化剂PTD-SOD对创伤愈合的影响.方法 制备机械性创伤小鼠模型和不同浓度的PTD-SOD(1 000 U、3 000 U、6 000 U、10 000 U)和SOD(3 000 U、6 000 U)溶液,分别用上述溶液进行治疗,同时设立模型对照组、等渗盐水对照组和复方碘液对照组,各组均连续治疗13天.观察各组创伤愈合情况,记录创伤收缩率和愈合天数;于创伤后第14天取各组小鼠创伤愈合部位皮肤,一部分制成10%组织匀浆液用于检测抗氧化酶(SOD、CAT、GSH-Px)活性及丙二醛(MDA)、羟脯氨酸(Hyp)含量,一部分制成病理组织切片用于皮肤组织学观察.结果 (1)与模型对照组、等渗盐水对照组及复方碘液对照组相比,同时相点PTD-SOD各组(10 000 U组除外)和SOD各组的抗氧化酶活性和Hyp含量显著升高(P<0.05或0.01),MDA含量显著降低(P<0.05或0.01),创伤收缩率提高(P<0.05或0.01),创伤愈合时间缩短(P<0.05或0.01).(2)在同等剂量下,从促创伤愈合时间、抗氧化酶活性、MDA含量、Hyp含量等方面比较,伤后同时相点PTD-SOD组明显优于SOD组(P<0.05或0.01).(3)10 000 U PTD-SOD组创伤愈合效果明显不如其他剂量PTD-SOD组(P<0.05或0.01).结论 在创伤愈合的早期特别是炎症期,应考虑氧化应激的存在,抗氧化治疗是一个治疗策略,适时、适当的抗氧化治疗,可以抑制氧化应激态,缩短创面愈合时间,提高创面愈合质量.适当剂量的PTD-SOD在皮肤创伤治疗中具有很好的抗氧化应激损伤、抑制炎症和促进愈合的效果.

Abstract：

Objective To study the role of the oxidative stress in the development of wound healing and observe the effect of the antioxidant PTD-SOD on damage and inflammation reaction after mechanical wound. Methods In this experiment,acute wound healing model by removal the whole layer dorsal skin of the mice was prepared,SOD(3 000 U and 6 000 U)and the fusion protein PTD-SOD with different concentrations(1 000 U,3 000 U,6 000 U and 10 000 U)were used to deal with the wounds continuously for 13 days.The mice were divided into different concentration SOD treatment group and PTD-SOD treatment group,model control group,physiological saline treatment group and compound iodine solution control group.The wound healing situation and healing percentage of the fight and left skin wounds of each mouse in every group was recorded every day.At day 14 after wound,the wound healing skin of each group was removed and some were used to make 10%tissues homogenate for detecting the activities of superoxide dismutase(SOD),catalase(CAT),glutathione peroxidase(GSH-Px)and contents of malondialdehyde(MDA)and hydroxyproline(Hyp);in the meantime,the other removed skin were fixed in 10% formalin for observing the histopathological changes of the tissues. Results Compared with the model control group,the physiological saline treatment group and the compound iodine solution control group,the skin wound healing percentage was significantly(P<0.05 or P<0.01)improved,with increase of the activities of SOD,CAT,GSH-Px and contents of Hyp (P<0.05 or P<0.01)and decrease of MDA(P<0.05 or P<0.01) in the SOD groups or PTD-SOD groups (except for 10 000 U PTD-SOD group).When compared with the physiological saline treatment group or the compound iodine solution treatment group,the effect was similar to the model control group.In comparison to the SOD groups,under the same dosage and environment condition,the PTD-SOD groups were much better than SOD groups with regard to promoting skin wound healing percentage,increasing activities of antioxidases and contents of Hyp,decreasing contents of MDA.Among the PTD-SOD groups,the effect of high dosage 10 000 U on promoting skin wound healing was declined. Conclusions The oxidative stress may playan important role in the development of wound healing.Proper application of treatment with antioxidants is a alternative strategy in the early stage of wound.PTD-SOD is able to prevent the oxidative stress damage,inhibit inflammatory infiltration and promote skin wound healing efficiently.     

局部应用PTD-SOD、SOD对小鼠皮肤创伤的抗氧化应激损伤保护效果及其比较

目的 探讨局部应用PTD-SOD、SOD对小鼠皮肤创伤的抗氧化应激损伤保护效果及其差异.方法 制备机械性创伤小鼠模型和不同浓度的PTD-SOD(1000、3000、6000 U)及SOD(1000、3000、6000 U)溶液,分别用上述溶液进行治疗,同时设立模型对照组和生理盐水对照组,各组均连续治疗13 d.观察各组创伤愈合情况,记录创伤愈合率和愈合天数;于创伤后第14天取各组小鼠创伤愈合部位皮肤,一部分制成10%组织匀浆液用于检测抗氧化酶(SOD、CAT、GSH-Px)活性及丙二醛(MDA)、羟脯氨酸(Hyp)含量,一部分制成病理组织切片用于皮肤组织学观察.结果 ①与模型对照组相比,PTD-SOD各组或SOD各组的抗氧化酶活性和Hyp含量显著(P<0.05)或极显著(P<0.01)升高,MDA含量显著(P<0.05)或极显著(P<0.01)降低,能显著(P<0.05)或极显著(P<0.01)提高创伤愈合率、缩短创伤愈合时间;与生理盐水对照组相比,结果类似.②在同等剂量下,从促创伤愈合时间、抗氧化酶活性、MDA含量、Hyp含量等方面比较,PTD-SOD组明显(P<0.05)或极明显(P<0.01)优于SOD组.③适当剂量的PTD-SOD促创伤愈合效果优于高剂量PTD-SOD的促创伤愈合效果.结论 PTD-SOD或SOD在皮肤创伤治疗中具有良好的抗氧化应激损伤效果,这种保护效果在创伤愈合的早期最显著;同等剂量下,PTD-SOD在促创伤愈合的效果上明显优于SOD.     

血府逐瘀胶囊对人微血管内皮细胞表达Dll4的影响

目的探讨血府逐瘀胶囊含药血清对体外培养的人微血管内皮细胞表达Dll4的影响。方法 12只SD大鼠随机分成血府逐瘀胶囊组和空白对照组,每组6只。分别制备血府逐瘀胶囊含药血清和空白血清。以2.5%浓度的血府逐瘀胶囊含药血清和空白血清分别干预人微血管内皮细胞-1(HMEC-1),通过实时定量PCR(Real-time PCR)检测药物处理细胞12,24,36,48 h时Dll4在mRNA水平的表达,通过蛋白免疫印迹(Western blot)检测药物处理细胞24,48,72 h时Dll4在蛋白质水平的表达。结果血府逐瘀胶囊含药血清干预内皮细胞36 h能上调Dll4在mRNA水平表达(P0.01),药物干预细胞48 h能同时上调Dll4在mRNA和蛋白水平表达。结论血府逐瘀胶囊很可能通过调控Dll4表达,影响相关信号通路,从而调控血管新生。