当归四逆汤对BALB/c硬皮病小鼠皮肤组织中CTGF,TGF-β含量的影响  

Effect of Danggui Sini Decoction on CTGF and TGF-β in Skin Tissue of BALB/c Mice with the Systemic Sclerosis

作　　者：王振亮[1] 宋建平[1] 张晓艳[1] 贾丽丽[1] 崔庆安[1] 

机构地区：[1]河南中医学院仲景医药研究所,郑州450008

出　　处：《中国实验方剂学杂志》2012年第23期179-182,共4页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81072716)

摘　　要：目的:观察当归四逆汤对BALB/c硬皮病小鼠皮肤组织及其结缔组织生长因子(connective tissue growth factor,CTGF)、转化生长因子(transforming growth factor,TGF)β含量的影响。方法:选8周龄近交系BALB/c小鼠60只,剃去背部中央区被毛,随机分为5组,每组12只。模型组和观察组小鼠背部皮内注射100μL(200 mg·L-1)博来霉素(BLM)溶液,每天1次,连续3周。正常组小鼠用磷酸盐缓冲液(PBS)0.1 mL做背部皮内注射。造模成功后,各组按20 mL·kg-1容量灌胃,连续3周。当归四逆汤高、中、低剂量组(39,23.4,7.8 g·kg-1)。正常组和模型组给等容量生理盐水。实验结束处死动物,切取小鼠背部注射区皮组织,部分剪碎后与生理盐水研磨成2%组织浆,3 000 r·min离心10 min,取部分上清液,用ELISA法测定CTGF,TGF-β的含量。部分组织固定于10%甲醛液,石蜡包埋,苏木素2伊红(HE)染色和Masson三色染色,分别用彩色病理图像分析系统和细胞免疫组织化学定量分析系统测定皮肤厚度和纤维化指数。结果:当归四逆汤高、中、低剂量均能减轻BLM致硬皮病小鼠的真皮厚度高、中剂量组分别为(25.22±2.35),(29.13±2.03)μm,P<0.01;低剂量组为(29.95±2.85)μm,P<0.05);高、中剂量组能降低硬皮病小鼠皮肤纤维化指数,分别为(86 615±8 403),(103 174±18 439),P<0.01;皮肤组织中CTGF分别为(160.2±35.7),(109.5±28.6)ng·L-1,P<0.01,TGF-β的含量分别为(48.48±4.95),(63.30±7.32)ng·L-1,P<0.01),而且存在明显的量效关系。结论:当归四逆汤能使模型小鼠的皮肤硬化得到改善,对其皮肤组织中的CTGF,TGF-β含量有降低作用。Objective： To observe the effect of Danggui Sini decoction on connective tissue growth factor （CTGF） and transforming growth factor-β （TGF-β） in skin tissue of the systemic sclerosis （SSc） mice of BALB/c. Method： BALB/c mice 60 with eight weeks old inbred strain were selected and the back fur of the central district was shaved off, and randomly divided into 5 groups （n = 12 each group）. The back skin of mouse in the model group and observation group was injected with 100 μL （200 mg · L^-1）, bleomycin （BLM） solution once each day for 3 weeks. The back skin of mouse in normal group was injected with phosphate buffered saline （PBS） 0. 1 mL. After the modeling, mice in each group was given 20 mL·kg^-1 by gavage daily for 3 weeks, the dosage of Danggui Sini decoction was 39, 23.4, 7.8 g ·kg^-1. Normal and model groups were given equal volume of saline, After the end of the experiment, the animals were killed, injected central skin was collected, andtissue homogenate was made with 2% saline and 3 000 r ·min^-1 centrifugation for 10 min, and then the supernatant was extracted and the content of CTGF and TGF-β were determined using ELISA method. Some of the skin was fixed in 10% formaldehyde solution, and then embedded in paraffin, and stained with hematoxylin-eosin （HE） and Masson staining, skin thickness and fibrosis index were determined, respectively, with color pathology images and Immunohistochemistry quantitative analysissystem. Result： Danggui Sini decoction could reduce the BLM induced scleroderma mouse dermal thickness （in high and middle dosage, which was 25.22 ± 2.35, .29.13 ± 2.03 respectively, P 〈0.01 while in the low dosage, 29.95 ~ 2.85, P 〈0.05. In high-dose group and the medium dose group skin fibrosis index was reduced （86 615 ± 8 403, 103 174 ± 18 439, respectively, P 〈 0.01 ） and skin tissue levels of CTGF （160.22 ± 35.73）, （109.51 ±28.64） ng · L^-1, respectively, P 〈 0. O1 ） and TGF- β （48.48 ±4.95）, （63.30 ±7.32�

关 键 词：硬皮病模型 当归四逆汤 小鼠 结缔组织生长因子 转化生长因子-β 

分 类 号：R285.5[医药卫生—中药学]