芹菜素对大鼠烧伤创面微循环改善作用的研究

Ameliorative Effect of Apigenin on Microcirculation of Burn Wounds in Rats

目的探讨芹菜素对大鼠烧伤创面微循环的改善作用及机制。方法180只雄性Wistar大鼠按随机数字表法随机分为假烧伤组、模型组、磺胺嘧啶银软膏单药组和磺胺嘧啶银软膏+芹菜素低、中、高剂量组,各30只,采用压力0.03MPa、106℃持续5s的方法制备深II度烧伤大鼠模型。假烧伤组和模型组腹腔注射0.9%氯化钠溶液2mL,每天1次;磺胺嘧啶银软膏单药组于烧伤皮肤涂抹磺胺嘧啶银软膏,每2天换药1次,腹腔注射0.9%氯化钠溶液2mL,每天1次;磺胺嘧啶银软膏+芹菜素低、中、高剂量组于烧伤皮肤涂抹磺胺嘧啶银软膏,每2天换药1次,腹腔注射相应浓度芹菜素溶液(12.5、25、50mg/kg),疗程14天。观察各组大鼠创面大体情况、HE染色法行创面组织病理学检查,计算烧伤创面愈合率,检测创面组织微循环血流灌注值(MPD)、微血管密度(MVD)和创面组织含水量,Western blot法检测表皮生长因子(EGF)、碱性成纤维细胞因子(b-FGF)、血管内皮生长因子(VEGF)蛋白表达量。结果与模型组比较,磺胺嘧啶银软膏+芹菜素低、中、高剂量组烧伤大鼠创面新生上皮生长活跃、病理性形态结构明显改善,创面愈合率增加[(61.09±6.85)%、(65.41±7.32)%、(71.70±8.74)%比(30.24±4.58)%,P均<0.01];创面组织MPD、MVD表达上调[MPD:(482.64±115.07)PU、(648.25±130.64)PU、(936.27±189.61)PU比(362.48±75.93)PU,P均<0.01;MVD:(24.17±1.75)piece/mm^2、(29.62±2.28)piece/mm^2、(34.16±2.53)piece/mm^2比(21.19±1.24)piece/mm^2,P均<0.01];创面组织EGF、bFGF、VEGF表达显著上调[EGF:(0.49±0.09)、(0.57±0.11)、(0.63±0.13)比(0.39±0.05),P均<0.01;bFGF:(0.21±0.06)、(0.39±0.09)、(0.52±0.14)比(0.15±0.02),P均<0.01;VEGF:(0.35±0.10)、(0.51±0.14)、(0.68±0.17)比(0.27±0.05),P<0.05或P<0.01];磺胺嘧啶银软膏+芹菜素中、高剂量组创面组织含水量显著低于模型组[(68.69±1.19)%、(68.27±1.20)%比(70.41±1.28)%,P均<0.01];与磺胺嘧啶银软膏单药组比较,磺胺嘧啶银软膏+芹菜素中、高剂量组创面组织病理性形态结构改变改善更明显,创面愈合率增加[(65.41±7.32)%、(71.70±8.74)%比(58.37±7.01)%,P<0.05或P<0.01];创面组织MPD、MVD表达上调[MPD:(648.25±130.64)PU、(936.27±189.61)PU比(445.35±90.18)PU,P均<0.01;MVD:(29.62±2.28)piece/mm^2、(34.16±2.53)piece/mm^2比(22.78±1.39)piece/mm^2,P均<0.01];创面组织EGF、bFGF、VEGF表达显著上调[EGF:(0.57±0.11)、(0.63±0.13)比(0.42±0.07),P均<0.01;bFGF:(0.39±0.09)、(0.52±0.14)比(0.17±0.04),P均<0.01;VEGF:(0.51±0.14)、(0.68±0.17)比(0.31±0.07),P均<0.01]。结论芹菜素能够通过改善大鼠深II度烧伤创面微循环而促进创面愈合,其机制可能与促进创面血管新生有关。

Objective To investigate the ameliorative effect and mechanism of apigenin on microcirculation of burn wound in rats.Methods One hundred and eighty male Wistar rats were randomly divided into sham burn group,model group,sulfadiazine silver ointment group,and sulfadiazine silver ointment plus low,medium and high dose of apigenin groups,each with 30 animals.The model rats with deep II degree burn injury were constructed by using a pressure of 0.03 MPa and 106℃for 5 seconds.The sham burn group and the model group received intraperitoneal injection of 2 mL of 0.9%sodium chloride solution once a day.Sulfadiazine silver ointment group was treated with sulfadiazine silver ointment to burned skin once every 2 days,and was intraperitoneally injected 2 mL of 0.9%sodium chloride solution once a day.sulfadiazine silver ointment plus low,medium and high dose of apigenin groups were treated with sulfadiazine silver ointment to burned skin once every 2 days,and were intraperitoneally injected apigenin solution with different concentration(12.5,25 and 50 mg/kg)for 14 days.The general condition of the wound surface of rats in each group was observed and histopathological examination of the wound surface was performed by HE staining.The healing rate of the burn wound surface was calculated.The microcirculation perfusion value(MPD),microvascular density(MVD)and water content of the wound surface were detected.Expressions of epidermal growth factor(EGF),basic fibroblast factor(b-FGF)and vascular endothelial growth factor(VEGF)were detected by Western blot.Results Compared to the model group,the neoepithelia of burned rats grew actively in sulfadiazine silver ointment plus low,medium and high dose of apigenin groups,the pathological morphological and structural changes were significantly improved,the wound healing rate increased[(61.09±6.85)%,(65.41±7.32)%and(71.70±8.74)%vs(30.24±4.58)%,P<0.01,respectively];MPD increased[(482.64±115.07),(648.25±130.64)and(936.27±189.61)vs(362.48±75.93)PU,P<0.01,respectively],MVD increased[(24.17±1.75),(29.62±2.28)and(34.16±2.53)vs(21.19±1.24)piece/mm^2,P<0.01,respectively];expression of EGF,bFGF and VEGF up-regulated,i.e.,EGF:[(0.49±0.09),(0.57±0.11)and(0.63±0.13)vs(0.39±0.05),P<0.01,respectively],bFGF:[(0.21±0.06),(0.39±0.09)and(0.52±0.14)vs(0.15±0.02),P<0.01,respectively],VEGF:[(0.35±0.10),(0.51±0.14)and(0.68±0.17)vs(0.27±0.05),P<0.05 or P<0.01].Water content in sulfadiazine silver ointment+medium-and high-dose of apigenin groups were lower than that in model group[(68.69±1.19)%and(68.27±1.20)%vs(70.41±1.28)%,P<0.01,respectively].Compared to sulfadiazine silver ointment group,the pathological morphological changes of wound tissues in the sulfadiazine silver ointment+medium-and high-dose apigenin groups improved more obviously,and the wound healing rate increased[(65.41±7.32)%and(71.70±8.74)%vs(58.37±7.01)%,P<0.05 or P<0.01];MPD increased[(648.25±130.64)and(936.27±189.61)vs(445.35±90.18)PU,P<0.01,respectively],MVD increased[(29.62±2.28)and(34.16±2.53)vs(22.78±1.39)piece/mm^2,P<0.01,respectively];expression of EGF,bFGF and VEGF up-regulated significantly,i.e.,EGF:[(0.57±0.11)and(0.63±0.13)vs(0.42±0.07),P<0.01,respectively],bFGF:[(0.39±0.09)and(0.52±0.14)vs(0.17±0.04),P<0.01,respectively],VEGF:[(0.51±0.14)and(0.68±0.17)vs(0.31±0.07),P<0.01,respectively].Conclusion Apigenin can promote wound healing by improving microcirculation of deep II degree burn wounds in rats,and its mechanism may be related to promoting wound angiogenesis.