IL-7 Antagonize The Stimulating Function Of TGF-β₁ To Fibroblast

PrefaceThe interstitial lung disease is a series of pulmonary fibrosis disease which pathogenesy is unclear and treatment is difficult. Its incidence and mortality rise gradually in recent years. No matter what is etiological factor and its pathogenesis, lung fibroblast is the common, final target cell. The TGF-β₁ is regarded as the most important cytofactor in beginning and development of fibrosis in many organs by most scholar at present. But the recent years research discovered idiopathic pulmonary fibrosis pathology characteristic-the fiber multiplication region is mainly composed of the myofibroblast expressingα-smooth muscle actin (α-SMA) and extracellular matrix such as collagen, moreover myofiobrosis is the main cell which makes the extracellular matrix abnormal deposited. Research prove that TGF-β₁ play the important role in this processes. Recently, Researcher had discovered IL-7 could decrease the generation of TGF-βin megakaryocytes, the fibrosarcoma cell and the melanian tumor cell. Therefore, base on the important function of TGF-βin pulmonary fibrosis, IL-7maybe have the anti- pulmonary fibrosis function through decreasing TGF-βgeneration.Therefore, this research's objective is to make it clear whether 11-7 can antagonize transformation from lung-fibroblast to myofibroblast stimulated by TGF-β₁, fibroblast multiplications and generation of collagen, which can help us understand the role of Il-7 in anti-pulmonary fibrosis and provides a new method for the lung fibrosis treatment.Research object and method1. Material Cell:The mouse embryo lung-fibroblast cell plant (BadB/c3T3, Shanghai China cell storehouse).Instrument:The refrigeractor, the centrifuge, the dispersion of light luminosiey meter, the western signature appliance, the thermostate, the microscope, the cell-culture laboratory bench and so on.Reagent:(1)hydroxyproline examination reagent box: Nanjing completes biological research institute.(2)Reorganization person TGF-β₁,reorganization mouse IL-7, peprotech company.(3)sole-clone mouse antia-smooth muscle actin antibody, Wuhan boster bro-engineering limited company, Goat anti-mouse two-anti, Wuhan boster bro-engineering limited company. (4)Embryo cow blood serum, buys to the Chinese Academy of medical Sciences bio-engineering research institute.(5)DMEM nutrient fluid. Hyclone company .(6)BCA protein density determination reagent box, Biyutian company.2. The experimental technique:After the fibroblast recover, stimulate them by TGF-β₁(concentration:10ng/ml), observe the situation of cell multiplication at 24,48,72 hours (MTT evaluate), the collagen generation (hydroxyproline digestion examination )andα-SMA expression (western blotting examination). Then, add different concentration IL- 7(50,100,150ng/ml) to interfere at the corresponding time, after 24 hours, reexamine them again.Result1.(50,100,150Different concentration ng/ml)IL-7 can inhibit the cell multiplication caused by TGF-β₁(10ng/ml) in different time (24,48,72h).2. 50ng/ml IL-7 has no obvious influence to collagen generation after TGF-β₁ (10ng/ml) stimulated fibroblast. 100ng/ml IL-7 has influence to collagen generation which can inhibit its production. 150ng/ml IL-7 has remarkable influence which can significantly inhibit collagen production3. Different concentration (50.100.150ng/ml)IL-7 can reducesα-SAM expression of fibroblast stimulated by TGF-β₁, its inhibitory effect depend on time and concentration.ConclusionThis article confirmed different concentration (50,100,150ng/ml)IL-7 can reduced cell multiplication, reduce collagen formation andα-SMA expression of fibroblast stimulated by TGF-β₁ in vitro.