青黛对结肠粘膜成纤维细胞的影响及作用机制-汕头科学技术协会.docVIP

青黛对结肠粘膜成纤维细胞的影响及作用机制 陈小璇, 代剑平(, 朱丹霞, 王革非, 李康生 汕头大学医学院 微生物与免疫学教研室，广东，汕头，515031 【摘要】目的：青黛临床治疗多种溃疡性疾病，本试验研究了青黛对结肠粘膜成纤维细胞的影响及其机制。方法：以不同方式提取青黛，并命名为水提物、醇提物和总提物，试验方法有羟脯氨酸测定，明胶酶谱分析，qRT-PCR，ELISA和Western blotting等。结果：水提物可促进结肠粘膜成纤维细胞增殖和胶原积累，提高MMPs活性和TIMP-1/-2生成，上调TGFβ1和CTGF水平，并激活PI3K/Akt、PKC及ERK/JNK/p38 MAPK信号通路。醇提物能抑制成纤维细胞增殖，减少TGFβ1、CTGF、IL-6、IL-1和TNF-α转录和释放，降低p-Akt和p-p38水平。总提物能促进成纤维细胞增殖和胶原积累，提高MMPs活性和TIMP-1/-2生成，上调TGFβ1和CTGF水平，激活PKC、ERK MAPK通路。结论：青黛水提物和醇提物对成纤维细胞增殖和胶原积累的作用相反，其总提物促进溃疡治愈可能依赖于PKC/ERK MAPK信号通路。 【关键词】青黛；成纤维细胞；溃疡愈合 Preliminary study of Indigo Naturalis on the functions of mice colonic fibroblasts CHEN Xiaoxuan, DAI Jianping, ZHU Danxia, WANG Gefei, LI Kangsheng Department of Microbiology and Immunology, Shantou University Medical College, Shantou 515041, China 【Abstract】 Objective To investigate the influence of indigo naturalis on the function of colonic fibroblasts relevant to ulcer healing. Methods Human procollagen gene promoter luciferase reporter plasmid, hydroxyproline assay, gelatin zymography, qRT-PCR, ELISA and western blotting were used. Results The aqueous extract could promote fibroblast proliferation and collagen accumulation, increased the production of MMPs, TIMP-1/-2, TGFβ1 and CTGF, and activated PI3K/Akt, PKC, ERK/JNK/p38 MAPK pathways. The alcoholic extract could inhibit fibroblast proliferation, decreased the production of TGFβ1, CTGF, IL-6, IL-1, TNF-α, and the levels of p-Akt and p-p38. The total extract could promote fibroblast proliferation and collage accumulation, increased the production of MMPs, TIMP-1/-2, TGFβ1 and CTGF, and activated PKC and ERK MAPK pathway. Conclusion Indigo naturalis contains two different kinds of ingredients with opposite effects for fibroblast proliferation and collage accumulation; the total extract promotes fibroblast proliferation and collage accumulation via activating PKC and ERK MAPK signal pathway. 【Key words】Indigo naturalis; Fibroblast; Ulcer healing 青黛是由菘蓝（Isatis indigotica）、板蓝根（Baphicacanthus cusi