新细胞因子-趋化素样因子（CKLF1）的心血管生物学效应

Chemokine-like factor 1 (CKLF1) is a novel cytokine isolated in professor Ma Dalong's laboratory, Peking University. It has chemotactic effect on a wide spectrum of cells both in vitro and in vivo; it can also stimulate the regeneration of skeletal muscle cells in vivo, but its biological effect on cardiovascular system remains unclear.Methods:(1) To construct the eukaryotic expression vector of hCKLF1;(2) To establish cell lines, including CKLF1-overexpressed stablely cell lines and a control cell lines (BALB/C 3T3-3T3 mouse fibroblast cells and A7r5-rat aortic smooth muscle cells);(3) RNA isolation and quantitative real-time RT-PCR;(4) Cell proliferation analyzed by MTT, cell counting assay, flow cytometry analysis;(5) Cell apoptosis analyzed by Annexin-V staining;(6) Immunoblot analysis of phospho-p38-MAPK;(7) Histological examination.Main contents:(1) In vitro: to observe the effects of hCKLF1 on proliferation and apoptosis of BALB/C 3T3 and A7r5;(2) In vivo: to observe the histological changes of important organs in BALB/C mouse with intravenous injection of hCKLF1-pcDI.Results and significances: (1) We have firstly successfully established cell lines, including CKLF1-overexpressed stablely cell lines and a control cell lines (BALB/C 3T3-3T3 mouse fibroblast cells and A7r5-rat aortic smooth muscle cells); (2) Overexpression of hCKLF1 promotes proliferation and survival of BALB/C 3T3 mouse fibroblast cells;(3) hCKLF1 could exert an anti-apoptotic effect on BALB/C 3T3 cells after serum withdrawal;(4) Overexpression of human CKLF1 promotes proliferation of A7r5 rat aortic smooth muscle cells through increasing level of intracellular phospho-p38-MAPK; These results provide further evidences for overlapping chemotactic effect and proliferation effect of cytokine, enriching cytokine theory.(5) In vivo, BALB/C mouse transfected with intravenous injection of hCKLF1, histological examination showed the proliferations of interstitial cell in the heart and vascular smooth muscle cell, which were consistent with cellular studies, but the degree of proliferation was less than cellular studies; no changes in myocardial cell structure; leucocytes (lymphocyte and neutrophil) infiltration in the lung tissue between bronchus and arteriole; white pulp in spleen became smaller; and proliferation of spermatogenic cell. The structure of other organs did not change. These results lay solid foundation for further investigating pathological effect of CKLF1 in the lung and heart.

已证明趋化素样因子（CKLF1）对多种细胞、骨髓和骨骼肌具有病理生理效应。预实验表明云交∠赴头涡《鲆灿忻飨宰饔谩＝徊酱犹迥谕庋芯緾KLF1的心血管变力、心肌细胞肥大效应，对平滑肌细胞、内皮细胞、成纤维细胞分化增殖的影响及心肌、冠状动脉、侧枝血管的结构改变特点，证实其心血管生物学效应，为开发新基因工程药物提供理论依据。