ABSTRACT
OBJECTIVE: To observe effect of Invigorating Lung and Kidney Prescription (ILKP) on secretion of cytokines induced by Lipopolysaccharide (LPS) and cigarette smoke extract (CSE) in vitro and discuss prescription regularity of ILKP and reveal molecular mechanism of ILKP treatment on COPD. METHODS: Alveolar epithelial cells A549, airway epithelial cells H292 and monocyte macrophage THP-1 were stimulated with LPS and CSE, in which samples were divided into normal group (20% normal rat serum), model group (20% normal rat serum and 5% CSE with 75 pg/mL LPS), ILKP group and its Components [20% Replenishing qi group (1), Replenishing Lung and Kidney group (2), Activating blood group (3), Reduce phlegm group (4), Replenishing qi with Replenishing Lung and Kidney group (5), Replenishing qi with Activating blood group (6), Replenishing qi with Reduce phlegm group (7), Replenishing qi with Replenishing Lung and Kidney and Activating blood group (8), Replenishing qi with Replenishing Lung and Kidney and Reduce phlegm group (9), Replenishing qi with Activating blood and Reduce phlegm group (10), ILKP serum group (11)]. The cytokines were detected by ELISA and the transcription factors of cytokines were analyzed with Network tool. RESULTS: Compared with the normal group,the secretion of MMP-9, GM-CSF and TGF-beta by THP-1 cells,TIMP by A549 cells,TNF-alpha, MMP-9, GM-CSF, CD36 and TIMP-1 by H292 cells all were increased, while secretion of TNF-alpha, PPAP2B, IL-3 and SOD decreased by THP-1 cells, MMP-9, TNF-alpha, TGF-beta and IL-3 by A549 cells, IL-8 by H292 cells all were decreased. Compared with model group, for THP-1 cells, the secretion of GM-CSF in 3, 6, 10 and 11 prescription groups,TGF-beta in 10 and 11 prescription groups all were decreased. MMP-9 decreased in 9 and 5 prescription groups while it increased in 1, 2, 3, 4, 7, 8, 10 and 11 prescription groups. TNF-alpha was increased in 1, 2, 3, 4, 6, 8, and 9 prescription groups, as it decreased in 11 prescription group. Except 2 prescription group, IL-3 was increased in all other drugs groups. SOD in 2, 4, 5, 8, 9, 11 prescription groups, and PPAP2B in 1, 2, 3, 4, 5, 6, 7 prescription groups were increased while PPAP2B was decreased in 11 prescription group. For A549 cells, the secretion of TIMP-1 was increased in 1, 2, 3, 4, and 5 prescription groups, while it was decreased in 11 prescription groups; Except 1 and 4 prescription group, MMP-9 was increased in other groups; Except TGF-beta was increased in 11 prescription groups, it was increased in other groups; IL-3 was increased in 1, 2, 4, 5, 11 and 3 prescription groups. For H292 cells, the secretion of GM-CSF in 5,6,7 and 2 prescription groups,TNF-alpha in 1, 2, 4, 6, 7, 8, 9 and 10 prescription groups, MMP-9 in 4, 8, 7 and 9 prescription groups all were decreased. CONCLUSION: The effect of ILKP and its components on secretion of cytokines induced by LPS and CSE and its target cells are different, in which Activating blood prescription focuses on inflammation cytokines, Replenishing Lung and Kidney prescription on Lipid metabolism and redox regulation. The effect of ILKPA is the most widely, followed by Activating blood Replenishing Lung and Kidney, and Reduce phlegm prescription on Protease regulation.
