摘要
目的:通过研究清热凉血中药对人永生化表皮细胞(Ha Ca T细胞)人β防御素(HBD)-2的表达及信号通路的调控作用,探讨其治疗银屑病的可能机制。方法:用白细胞介素(IL)-1β和肿瘤坏死因子(TNF)-α的混合物刺激Ha Ca T细胞建立信号通路活化及HBD-2高表达的Ha Ca T细胞模型;分别用清热凉血中药、维A酸和生理盐水进行干预,用实时荧光定量核酸扩增法(q PCR)和酶联免疫吸附试验(ELISA)测定清热凉血中药对细胞模型中HBD-2表达水平的影响;用免疫印迹法(westen blot)检测清热凉血中药对细胞模型中HBD-2及丝裂原活化蛋白激酶(mitogen-activated protein kinases,MAPK)和核转录因子kappa B(NF-κB)信号通路的影响。结果:(1)中药血清高剂量组和维A酸血清组均可明显下调Ha Ca T细胞模型中HBD-2m RNA的表达,抑制率分别为66.8%和73.5%,与细胞模型组比较,差异均有统计学意义(均P<0.01),中药血清高剂量组和维A酸血清组比较差异无统计学意义(P>0.05);(2)中药血清高剂量组与维A酸血清组均可抑制细胞培养上清中HBD-2的分泌,与细胞模型组比较,抑制率分别为35.6%和61.6%;(3)100 mg/L的IL-1β和TNF-α混合物作用18 h后可使Ha Ca T细胞中的cJun氨基末端激酶(JNK)2 MAPK信号通路活化;清热凉血中药血清可以明显下调磷酸化c-Jun氨基末端激酶(p-JNK)2蛋白的相对含量,抑制JNK2 MAPK信号通路的活化。结论:清热凉血方药可明显下调HBD-2高表达的Ha Ca T细胞模型及培养上清中HBD-2的表达。清热凉血方药可以抑制JNK2 MAPK信号通路,并阻断IL-1β和TNF-α对HBD-2的激活。
Objective： To study the effects of heat-clearing and blood-cooling medicines on HBD-2 expression and JNK2/ MAPK signaling pathway in order to elucidate the mechanisms by which these herbal medicines benefit psoriasis. Method： HaCaT cells were stimulated with a mixture of TNF-α and IL-1β to increase HBD-2 expression and to activate JNK2/MAPK signaling pathway, followed by addition of serum containing either heat-cleaving and blood-cooling medicines, retinoic acid, or normal saline. The expression levels of HBD-2, NF-KB, p38 MAPK and JNK1 were assessed using ELISA, real-time PCR and western blot techniques. Result： Heat-clearing and blood-cooling medicines and retinoic acid exhibited a similar inhibitory ef- fect on HBD-2 mRNA expression（66.8% inhibition in heat-clearing and blood-cooling medicines-treated and 73.5% in retinoic acid-treated; P〈0.01 vs. vehicle for both treatments）. Moreover, heat-clearing and blood-cooling medicines and retinoic acid in- hibited the expression of HBD-2 protein by35.6% and 61.6%, respectively. While JNK2/MAPK signaling pathway was activated following the treatment of HaCaT cells with 100mg/L of IL-1β and TNF-α for 18h, heat-clearing and blood-cooling medicines significantly decreased the levels of p-JNK2 protein in cells treated with IL-1β and TNF-α, indicating a inhibition of JNK2/ MAPK signaling pathway. Conclusion： Heat-clearing and blood-cooling medicines can markedly decrease the expression of HBD-2 via inhibition of JNK2/MAPK signaling pathway, and inhibit IL-1β and TNF-α mediated activation of HBD-2.
出处
《临床皮肤科杂志》
CAS
CSCD
北大核心
2016年第9期625-629,共5页
Journal of Clinical Dermatology
关键词
银屑病
MAP激酶信号系统
NF-κB
Β防御素
清热凉血方药
psoriasis
MAP kinase signaling system
NF-kappa B
beta-defensins
heat-clearing and blood-cooling recipe
