愈创蓝油烃促进皮肤创伤修复的研究

Research on guaiazulenum treatment of skin wound

为了探究愈创蓝油烃促进创伤愈合的机制,选取40只SD大鼠制作皮肤创伤模型,随机分为蓝油烃治疗组和模型对照组。于造模后第3、7、14、21天各处死5只大鼠,用ImageJ软件测量创面面积,计算愈合率。用Masson染色观察伤口胶原沉积量,用qRT-PCR检测VEGF、TGF-β1、FGF-7、EGF、HGF、Smad2和Smad3的转录水平,用免疫组织化学方法检测创面微血管再生情况,用ELISA检测IL-1α、MPO、PGE-2和TNF-α的表达水平。结果显示,愈创蓝油烃组的创伤愈合率高于模型对照组的。在用药后第3天,用qRT-PCR检测相关生长因子,结果显示,愈创蓝油烃组EGF的表达量显著低于对照组的,TGF-β1的表达量与对照组的无明显差异;第7天,Smad2的表达量与对照组的无明显差异;其余各时间点,愈创蓝油烃组VEGF、HGF、FGF-7、Smad3的转录水平均高于对照组的。由Masson染色结果可见在创伤愈合的整个过程中,蓝油烃处理组大鼠的胶原沉积率高于对照组的。由免疫组织化学检测结果可见在第3和7天时,蓝油烃处理组的创面微血管密度高于对照组的,第14天时则低于对照组的。ELISA结果显示第3和7天时,愈创蓝油烃组IL-1α含量低于对照组的,而第14天时略高于对照组的;于第3、7天时,愈创蓝油烃组MPO含量较对照组的略有下降,但在第14天时略高于对照组的;各时间点治疗组PGE-2水平均低于对照组的;治疗组大鼠TNF-α水平在第3天时略高于对照组的,在第7和14天时愈创蓝油烃组低于对照组的。结论:愈创蓝油烃可促进大鼠皮肤创伤愈合。其作用机制可能是通过调控伤口胶原蛋白的沉积、抑制IL-1α和MPO等炎症因子的表达和上调TGF-β1与FGF-7等生长因子的表达等促进伤口愈合。

In order to explore the mechanism of guaiazulenum promoting wound healing,40 SD rats were selected to make skin wound models,and they were randomly divided into guaiazulenum treatment group and model control group.Five rats were killed on the 3 rd,7 th,14 th,and 21 st day of the wound and the wound area was measured with Image J software and the healing rate was calculated.Masson staining was used to observe wound collagen deposition.q RT-PCR was used to detect VEGF,TGF-β1,FGF-7,EGF,HGF,Smad2,and Smad3 transcription levels,immunohistochemistry was used to measure wound microvascular regeneration;ELISA was used to measure the expression levels of IL-1α,MPO,PGE-2,TNF-α;Western-blot was used to detection of TGF-β1,VEGF expression level.From the perspective of wound healing rate,the wound healing rate of guaiazulenum was higher than that of the model group.The q RT-PCR results showed that:on the 3 rd day,the expression of EGF in the treatment group was significantly lower than that in the control group,and the expression of TGF-β1 in the treatment group did not change significantly from that in the control group.On the 7 th day,the expression of Smad2 in the treatment group did not change much from that in the control group.At other time points,the transcription level of the treatment group was higher than that of the control group.Masson staining results showed that during the whole process of wound healing,the collagen deposition rate of the guaiazulenum treatment group was higher than that of the control group.The results of immunohistochemistry showed that on the 3 rd and 7 th day,the microvessel density of the wound in the guaiazulenum treatment group was higher than that of the control group,and it was lower than that of the control group on the 14 th day.ELISA results showed that the IL-1αcontent of the treatment group was lower than that of the control group on the 3 rd and 7 th day,and slightly higher than that of the control group on the 14 th day;on the 3 rd and 7 th day,the MPO content of the treatment group was slightly lower than that of the control group,and slightly higher that of control group on the 14 th day;PGE-2 levels in the treatment group were lower than those of the control group at each time point;the TNF-αlevel of rats in the treatment group was slightly higher on the 3 rd day,and the treatment level was lower than that of the control on the 7 th and 14 th day.In conclusion:Guaiazulenum can promote the healing of rat skin wounds.Its mechanism is to regulate the deposition of wound collagen,inhibit IL-1α,MPO and other inflammatory cytokines,and up-regulate the expression of growth factors such as TGF-β1 and FGF-7 to promote wound healing.