Study On The Anti-inflammatory Effect And Mechanism Of R-Salbutamol

Racemic Salbutamol is a β₂ adrenergic agonist which have anti-inflammatory effect,which commonly used in the treatment of bronchial asthma,It is an exact 50:50 mixture of two enantiomers,However,there was less systematic reports and lots of controversies to describe the inflammatory effect using different compounds.To address this controversy and to further clarify the effects of Salbutamol on inflammation.We have to analyze the anti-inflammatory mechanism in vivo and in vitro which lead to clinic treatment.This paper analyzes the different isomers of salbutamol,determination of the optical purity and chemical purity.We verified the S-salbutamol has proinflammatory effect in macrophages and we also verified the R-salbutamol elicits anti-inflammatory effects in ACD mice in vivo and lipopolysaccharide-induced inflammatory responses in vitro.Methods and results are as follows:1.This paper firstly analysis the different isomers of salbutamol and determine the optical purity and chemical purity and effects of R-Sal and S-Sal on Raw264.7.The results showed that,different concentrations of R-Sal and S-al has no inhibition to murine RAW264.7 cells growth,compared to the control group,there was no significant difference（p> 0.05）.S-Sal can increase the expression of NO,ROS,TNF-α,and R-Sal did not influence on RAW264.7.2.The present study investigate the anti-inflammatory effects R-salbutamol on mice with allergic contact dermatitis.ACD in mice was induced by the repeated application of 2,4-dinitrochlorobenzene（DNCB）to their skins.And we also contrast different concentrate R-Salbutamol and indomethacin to mice ACD model by morphology,pathological observation and molecular analysis level.The data shows medium concentrate R-Salbutamol has better function than indomethacin with dose-response relationship.The result is salbutamol has anti-inflammatory effects in vivo.3.This paper studied the anti-inflammatory effect of R-Sal on LPS induced RAW264.7 cells in vitro.Results show that: R-Sal can significantly reduce the LPS induced production of inflammatory factors in all kinds of content,and between the different concentrations was dose-response relationship,R-Sal inhibits of inflammatory expression in vitro,with R-Sal instead of Racemic Salbutamol and clinical application is reasonable.4.This paper also preliminary clarify the anti-inflammation mechanism about R-Salbutamol.Using β₂ adrenergic receptor agonist（ICI118551）Inhibit the function of R-Salbutamol when the 100 ng/m L LPS induce the inflammation in RAW264.7 cell.Comparing the fluorescence intensity of NO and ROS,the data shows the Salbutamol with ICI can not anti-inflammatory action（P < 0.01）.This result tells the mechanism of anti-inflammatory action is Salbutamol is β₂ adrenergic receptor agonist.And the RT-PCR was used to measure PDE4B mRNA level in different group cells.The data shows R-Salbutamol can decrease the level of intracellular PDE4B（P < 0.01）.PDE4B is one of phosphodiesterase which can make c AMP break up into AMP.Salbutamol can inhibits PDE4B which can reduce the phosphodiesterase activity.And the split of c AMP was inhibited and the intracellular quantity of c AMP was higher which reduce inflammation.The result is Salbutamol is β₂ adrenergic receptor agonist even can be anti-inflammatory drug which reduce intracellular PDE4B.