复方青黛丸联合MTX治疗银屑病疗效及机制研究

doi:10.3969/j.issn.1673-8640.2020.02.006

摘要/Abstract

摘要：

目的研究中成药复方青黛丸联合甲氨喋呤（MTX）对银屑病患者辅助性T细胞（Th）1、Th2细胞免疫平衡调节机制,评价联合用药的疗效。方法选取50例银屑病皮损面积和严重程度评分（PASI）>10的斑块型银屑病患者,随机分为2个组,每组25例患者。一组服用MTX,每周1次;另一组服用复方青黛丸联合每周1次的MTX治疗,为期8周。收集患者治疗前（基线）和治疗8周后血样,分离外周血单个核细胞（PBMC）,应用实时聚合酶链反应（PCR）分析Th1转录因子T-bet,细胞因子白细胞介素（IL）-12和干扰素-γ（IFN-γ）,以及Th2转录因子GATA- 3和细胞因子IL-4的表达。临床表现通过PASI评分进行评估。结果治疗至第8周时,复方青黛丸联合MTX治疗组有12例患者达到PASI-75（定义为PASI评分降低75%或更多）,单用MTX组有8例患者达到PASI-75,2个组临床表现差异有统计学意义（P<0.05）。复方青黛丸联合MTX治疗组Th1相关细胞因子下调,Th2相关细胞因子上调（P<0.05）。进一步以PASI-75为阈值,复方青黛丸联合MTX治疗组达到PASI-75者的IFN-γ表达比单用MTX组患者下降更为明显（P<0.05）;复方青黛丸联合MTX治疗组达到PASI-75的患者GATA-3和IL-4水平明显高于单用MTX组患者（P<0.05）。结论复方清黛丸联合MTX治疗对银屑病有更好的疗效,其原因可能是联合治疗对Th1、Th2细胞具有更强的免疫调控作用。

关键词: 细胞因子, 基因表达, 复方青黛丸, 甲氨喋呤, 银屑病

Abstract:

Objective To study the regulation of Chinese herbal medicine compound Qingdai capsule combined with methotrexate（MTX） on the immune balance of helper T cell（Th）1 and Th2 in patients with psoriasis,and to evaluate the efficacy of combined therapy. Methods A total of 50 patients with plaque psoriasis with a psoriasis area and severity index（PASI）>10 were enrolled. The study lasted for 8 weeks,of which 25 patients received weekly MTX treatment,and the other 25 patients received daily oral compound Qingdai capsule combined with weekly MTX treatment. Blood samples were collected before treatment（baseline） and after 8 weeks of treatment. Peripheral blood mononuclear cell（PBMC） was isolated,and real-time polymerase chain reaction（PCR） was used to analyze Th1 transcription factor T-bet,cytokine interleukin（IL）-12 and interferon-gamma（IFN-γ）. In addition,the Th2 transcription factor GATA-3 and cytokine IL-4 were also determined. Clinical performance was assessed by PASI. Results At the 8th week of treatment,12 patients in compound Qingdai capsule combined with MTX group achieved PASI-75（defined as at least 75% reduction in PASI）,and 8 patients in single MTX group achieved PASI-75. There was statistical significance in clinical manifestation between the 2 groups（P<0.05）. The Th1-related cytokines were down-regulated,and the Th2-related cytokines were up-regulated in compound Qingdai capsule combined with MTX group（P<0.05）. Furthermore,with PASI-75 as the threshold,in compound Qingdai capsule combined with MTX group,the IFN-γ expression of PASI-75 was lower than that in single MTX group（P<0.05）. Compound Qingdai capsule combined with MTX group reached PASI-75,and GATA-3 and IL-4 levels were higher than those in single MTX group（P<0.05）. Conclusions Compound Qingdai capsule combined with MTX has better therapeutic effect on psoriasis. The reason may be that combination treatment has strong immune regulatory effect on Th1 and Th2.

Key words: Cytokine, Gene expression, Compound Qingdai capsule, Methotrexate, Psoriasis

中图分类号:

R446.1

季天皓, 陈军毅, 孟亚东. 复方青黛丸联合MTX治疗银屑病疗效及机制研究[J]. 检验医学, 2020, 35(2): 120-124.

JI Tianhao, CHEN Junyi, MENG Yadong. Therapeutic effect and mechanism of compound Qingdai capsule combined with MTX in the treatment of psoriasis[J]. Laboratory Medicine, 2020, 35(2): 120-124.

图/表 6

参考文献 15

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基因	编号	引物序列（5'→3'）
IL-4	AM937235	上游:CGGCTCGACAGGAACCTCT
		下游:TCCAAGAAGTTTTCCAACGTACTCT
IL-12	AF101062	上游:GAGGGCCGTCAGCAACAT
		下游:ATGATCAATCAATCTCTTCAGAAGTGCAA
GATA-3	NM_002051	上游:TTCCCCAAGAACAGCTCGTT
		下游:GGCTCAGGGAGGACATGTGT
T-bet	AF241243	上游:ACACGCATATCTTTACTTTCCAAGAA
		下游:TCAGCTGAGTAATCTCGGCATTC
IFN-r	BC0701256	上游:CTAATTATTCGGTAACTGACTTGA
		下游:ACAGTTCAGCCATCACTTGGA
GAPDH	NM_001256799	上游:AGCCGGGCATGTTCTTCAAC
		下游:AGGGAGCTTCACGTTCTTGTAT

基因	编号	引物序列（5'→3'）
IL-4	AM937235	上游:CGGCTCGACAGGAACCTCT
		下游:TCCAAGAAGTTTTCCAACGTACTCT
IL-12	AF101062	上游:GAGGGCCGTCAGCAACAT
		下游:ATGATCAATCAATCTCTTCAGAAGTGCAA
GATA-3	NM_002051	上游:TTCCCCAAGAACAGCTCGTT
		下游:GGCTCAGGGAGGACATGTGT
T-bet	AF241243	上游:ACACGCATATCTTTACTTTCCAAGAA
		下游:TCAGCTGAGTAATCTCGGCATTC
IFN-r	BC0701256	上游:CTAATTATTCGGTAACTGACTTGA
		下游:ACAGTTCAGCCATCACTTGGA
GAPDH	NM_001256799	上游:AGCCGGGCATGTTCTTCAAC
		下游:AGGGAGCTTCACGTTCTTGTAT

组别	例数	性别		年龄/岁	体质量/kg	BMI/ （kg/m²）	PASI		DLQI
组别	例数	男性	女性	年龄/岁	体质量/kg	BMI/ （kg/m²）	治疗前	治疗后	治疗前	治疗后
对照组	25	17	8	37.8 （25.3,49.2）	66.5±6.38	31.03±4.97	11.03±1.97	5.61±0.73	11.35 （7.65,14.02）	7.35 3.65,8.02）
治疗组	25	15	10	37.5 （27.1,48.2）	65.9±10.11	30.90±5.27	10.90±1.27	7.06±0.84	11.28 （7.49,13.32）	6.35 （3.15,7.02）
P值		0.87		0.93	0.92	0.89	0.70	0.04	>0.05	>0.05

组别	例数	性别		年龄/岁	体质量/kg	BMI/ （kg/m²）	PASI		DLQI
组别	例数	男性	女性	年龄/岁	体质量/kg	BMI/ （kg/m²）	治疗前	治疗后	治疗前	治疗后
对照组	25	17	8	37.8 （25.3,49.2）	66.5±6.38	31.03±4.97	11.03±1.97	5.61±0.73	11.35 （7.65,14.02）	7.35 3.65,8.02）
治疗组	25	15	10	37.5 （27.1,48.2）	65.9±10.11	30.90±5.27	10.90±1.27	7.06±0.84	11.28 （7.49,13.32）	6.35 （3.15,7.02）
P值		0.87		0.93	0.92	0.89	0.70	0.04	>0.05	>0.05

组别	例数	Th1通路						Th2通路
		IFN-γ mRNA		IL-12 mRNA		T-bet mRNA		IL-4 mRNA		GATA3 mRNA
		PASI≤75%	PASI>75%	PASI≤75%	PASI>75%	PASI≤75%	PASI>75%	PASI≤75%	PASI>75%	PASI≤75%	PASI>75%
对照组	25	6.78±1.12	4.36±0.13	7.58±1.35	5.71±1.87	3.48±1.05	3.57±1.88	1.93±0.76	1.98±0.66	11.69±2.33	13.17±1.23
治疗组	25	4.59±0.21	2.51±0.12	5.38±0.87	4.85±0.44	1.71±0.89	1.63±0.45	2.55±0.13	2.11±0.12	18.92±2.36	20.13±1.58
P 值		0.04	0.03	0.04	0.88	0.31	0.17	0.05	0.05	0.62	0.05