龙胆苦苷对溃疡性压疮模型大鼠创面愈合的影响

Effect of gentiopicroside on wound healing in rat models with ulcerative pressure ulcer

目的观察龙胆苦苷对溃疡性压疮模型大鼠创面愈合的影响。方法将48只SD雄性大鼠随机分为正常对照组、模型组、养阴生肌散组及12.5 mg龙胆苦苷组、25 mg龙胆苦苷组、50 mg龙胆苦苷组,每组8只。除正常对照组外,其余组大鼠进行溃疡性压疮造模。造模后,养阴生肌散组及龙胆苦苷各组压疮部位均匀撒敷药粉,均2次/d,无菌敷料纱布遮盖,干预15 d。记录各组干预3 d、7 d、11 d、15 d后创面面积,比较各组干预15 d后创面愈合率,统计各组创面愈合时间。干预15 d后,取创面骨骼肌组织,采用HE染色进行形态学观察,免疫组化及RT-PCR检测增殖细胞核抗原(PCNA)蛋白及mRNA表达水平。结果养阴生肌散组及龙胆苦苷各组干预3 d、7 d、11 d、15 d后创面面积均显著低于同期模型组(P均<0.05),且50 mg龙胆苦苷组干预15 d后创面面积显著低于同期养阴生肌散组(P<0.05)。干预15 d后,养阴生肌散组及龙胆苦苷各组创面愈合率均显著高于模型组(P均<0.05),且50 mg龙胆苦苷组创面愈合率显著高于养阴生肌散组(P<0.05)。养阴生肌散组及龙胆苦苷各组创面愈合时间均显著短于模型组(P均<0.05),且50 mg龙胆苦苷组显著短于养阴生肌散组(P<0.05)。HE染色显示养阴生肌散组及龙胆苦苷各组创面组织炎性细胞明显减少,肌细胞增殖增加;RT-PCR及免疫组化结果显示,养阴生肌散组及龙胆苦苷各组创面骨骼肌组织中PCNA蛋白及mRNA表达水平均显著高于模型组(P均<0.05),且50 mg龙胆苦苷组增高明显,但与养阴生肌散组比较差异均无统计学意义(P均>0.05)。结论龙胆苦苷是养阴生肌散治疗溃疡性压疮的主要有效成分,可能通过上调创面中PCNA的表达促进肌细胞增殖,减少创面组织炎性细胞,从而促进创面愈合。

Objective It is to observe the effect of gentiopicroside on wound healing in rat models with ulcerative pressure ulcer.Methods Forty-eight SD male rats were randomly divided into normal control group,model group,Yangyin Shengji Powder group,12.5 mg gentiopicroside group,25 mg gentiopicroside group,and 50 mg gentiopicroside group,8 rats in each groups.Except for the normal control group,the rats in the other groups were modeled for ulcerative pressure ulcers.After modeling,the pressure ulcers of the Yangyinshengji Powder group and the gentiopicroside groups were evenly sprinkled with the drug powder for application,2 times per day,covered with aseptic dressing gauze,the intervention lasted for 15 days.The wound area of each group after 3,7,11,15 days of intervention was recorded,the wound healing rate of each group after 15 days of intervention was compared,and the wound healing time of each group was counted.After 15 days of intervention,the skeletal muscle tissue of the wound was taken for morphological observation by HE staining,and the expression levels of proliferating cell nuclear antigen(PCNA)protein and mRNA were detected by immunohistochemistry and RT-PCR.Results The wound area of the Yangyin Shengji Powder group and the gentiopicroside groups after intervention for 3 d,7 d,11 d,and 15 d were significantly lower than that of the model group at the same time(all P<0.05),and the area of the 50 mg gentiopicroside group intervention for 15 d was significantly lower than that of the Yangyin Shengji Powder group during the same period(P<0.05).After 15 days of intervention,the wound healing rates of the Yangyin Shengji Powder group and the gentiopicroside groups were significantly higher than that of the model group(P<0.05),and the rate of the 50 mg gentiopicroside group was significantly higher than that of the Yangyin Shengji group(all P<0.05).The wound healing time of the Yangyin Shengji Powder group and the gentiopicroside groups was significantly shorter than that of the model group(all P<0.05),and the time in the 50mg gentiopicroside group was significantly shorter than that in the Yangyin Shengji Powder group(P<0.05).HE staining showed that the inflammatory cells in the wound tissue of the Yangyin Shengji Powder group and the gentiopicroside groups were significantly reduced,and the proliferation of muscle cells was increased;RT-PCR and immunohistochemical results showed that the expression levels of PCNA protein and mRNA in the wound skeletal muscle tissues of Yangyin Shengji Powder group and gentiopicroside groups were significantly higher than that of the model group(all P<0.05),and the increase in the 50 mg gentiopicroside group was more significant,but there was no statistical difference compared with the Yangyin Shengji Powder group(all P>0.05).Conclusion Gentiopicrin is the main effective component of Yangyin Shengji Powder in the treatment of ulcerative pressure ulcers.It may increase the expression of PCNA in the wound to promote the proliferation of muscle cells and reduce the inflammatory cells in the wound tissue,thereby promoting wound healing.