马齿苋提取物对急性湿疹大鼠皮肤HRH1和Filaggrin基因表达的影响

发布时间：2018-06-11 19:04

本文选题：急性湿疹 + 马齿苋鲜榨液　；参考：《成都中医药大学》2016年硕士论文

【摘要】：目的：观察两种不同马齿苋提取物有效成分对急性湿疹大鼠背部皮肤组织HRH1和Filaggrin基因表达的影响,探讨不同马齿苋提取物治疗急性湿疹的可能机制。方法：SD大鼠50只,♂早各半,随机分为5组,每组10只,即：正常组、模型组、阳性药物对照组、马齿苋醇提取物组、马齿苋鲜榨液组。除正常组外,用电剃须刀剃去其余四组大鼠腹部毛发,面积约1 cm×2 cm,并在大鼠剃毛部位涂抹5%的2,4-二硝基氯苯(DNCB) 30μl于皮肤首次致敏;在致敏后第三天,再用电剃须刀剃去左、右对称背部约为2cm×2cm面积的剃毛区域,并在右背部剃毛区涂DNCB进行二次抗原攻击,浓度为0.2%,用量为50μ1,左背部对称的剃毛区不用药物,作为对照部位。以后按上述方法连续激发三次,每三天一次,用药后第九天观察大鼠右背部皮肤明显出现红斑,水肿,丘疹,糜烂或渗出,提示造模成功。于激发后第十天开始给药,每天两次,每次1ml,连续九天,最后在首次致敏后的第二十一天断颈处死大鼠,剪取造模区皮肤,分别予RT-PCR检测各实验组大鼠皮肤Filaggrin基因表达和免疫组化检测HRH1平均光密度。结果：1、与正常组相比,模型组大鼠皮肤组织HRH1蛋白含量明显升高,具有非常显著的统计学意义(P0.01),与模型组相比,马齿苋鲜榨液组大鼠皮肤组织HRH1蛋白含量明显降低,具有非常显著的统计学意义(P0.01),阳性对照组、马齿苋醇提物组大鼠皮肤组织HRH1蛋白含量降低,具有统计学意义(P0.05)。2、与正常组相比,模型组Filaggrin基因的2~(-△△CT)值极显著小(P0.01),与模型组比较,马齿苋鲜榨液组和阳性对照组大鼠皮肤组织细胞中Filaggrin基因的2~(-△△CT)值极显著增大(P0.01),马齿苋醇提取物组大鼠皮肤组织细胞中Filaggrin基因的2~(-△△CT)值极显著增大(P0.05)。结论：两种马齿苋提取物都可以使血管收缩,抑制毛细血管通透性,抑制炎性肿胀,从而减轻湿疹局部的瘙痒、水肿症状。尤其是马齿苋鲜榨液效果更好。其机制可能是两种马齿苋提取物能够显著的降低大鼠皮肤组织中瘙痒因子HRH1的水平,明显控制大鼠搔抓反应和改善了皮损情况,同时两种马齿苋提取物上调湿疹动物皮肤屏障系统中Filaggrin基因的表达,加速Filaggrin水解成各种天然保湿因子,有效改善和修复大鼠皮肤屏障功能,增强了机体的免疫功能,从而起到了治疗急性湿疹的作用。
[Abstract]:Objective: to observe the effects of two effective components of purslane extract on the expression of HRH1 and Filaggrin gene in the dorsal skin of rats with acute eczema, and to explore the possible mechanism of different extracts of Portulaca oleracea for the treatment of acute eczema. Methods Fifty Sprague-Dawley rats were randomly divided into 5 groups: normal group, model group, positive drug control group, ethanol extract group, fresh extract group of Portulaca oleracea and fresh extract of Portulaca oleracea. In addition to the normal group, the abdominal hair of the other four groups was shaved with electric razor, the area of which was about 1 cm 脳 2 cm, and the skin was sensitized with 5% of DNCB30 渭 l on the shaving area of the rats, and the left was shaved with the razor on the third day after sensitization. The right symmetrical back is about the shaving area of 2cm 脳 2cm, and the second antigenic attack is carried out in the shaving area of the right back. The concentration is 0.2and the dosage is 50 渭 1. The symmetrical shaving area of the left back is not used as the control site. The results showed that the skin of the right back of the rats appeared erythema, edema, papules, erosion or exudation on the ninth day after treatment, indicating that the model was successful. On the tenth day after stimulation, the rats were given drugs twice a day, 1ml / time for nine days. Finally, the rats were killed at the 21 day after the first sensitization, and the skin of the model area was cut off. The expression of Filaggrin gene was detected by RT-PCR and the mean optical density of HRH1 was detected by immunohistochemistry. Results compared with the normal group, the content of HRH1 protein in the skin tissue of the model group was significantly higher than that of the normal group, and the content of HRH1 protein in the skin tissue of the model group was significantly lower than that of the model group, and the content of HRH1 protein in the skin tissue of the model group was significantly lower than that of the model group. The content of HRH1 protein in the skin of rats in the positive control group and the ethanol extract group of Portulaca oleracea was significantly lower than that in the normal group, and the content of HRH1 protein in the skin tissue of the rats was significantly lower than that in the normal group. The value of Filaggrin gene in the model group was significantly lower than that in the model group (P 0.01), and was significantly lower than that in the model group. The value of Filaggrin gene in skin tissue cells of fresh extract group and positive control group was significantly increased (P 0.01), and that of Filaggrin gene in skin tissue cells of purslane alcohol extract group was significantly increased (P 0.05). Conclusion: both extracts of Portulaca oleracea can induce vasoconstriction, inhibit capillary permeability, inhibit inflammatory swelling, and relieve local itching and edema in eczema. Especially the fresh extract of Portulaca oleracea was better. The mechanism may be that the two extracts of Portulaca oleracea can significantly reduce the level of pruritus factor HRH1 in the skin tissue of rats, control the scratching reaction of rats and improve the condition of skin lesions. At the same time, two extracts of Portulaca oleracea upregulated the expression of Filaggrin gene in the skin barrier system of eczema animals, accelerated the hydrolysis of Filaggrin into various natural moisturizing factors, effectively improved and repaired the skin barrier function of rats, and enhanced the immune function of the body. Thus played a role in the treatment of acute eczema.
【学位授予单位】：成都中医药大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R285.5

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