The Experimental Study On The Regulation Of NF-Kappa B And TGF-Beta Signaling Pathway In Macrophages By Pseudo-ginseng And Zedoary

Objective:Based on the clinical experience of the famous traditional Chinese medicine in Yunnan,the prescription of Yifu Sanjie Fang(Also called Yifu Zhuyu Fang)has a good effect on alleviating airway inflammation and improving COPD airway remodeling.This experiment further studies the effect of zedoary and Pseudo-ginseng active components main components curcumol,curcumin,total saponins of Panax notoginseng on macrophages in this prescription,and explores whether the active components can reduce inflammatory factors by inhibiting the NF-kappa B signaling pathway to alleviate inflammation and detect the related indexes of TGF-beta /Smads signaling pathway,and explore three activities in the cell damage model.The composition acts on the relationship between the two signal pathways.Methods:The Raw264.7 cell damage model was established by using cigarette extract(Cigarette smoke extract,CSE)to stimuLate the cells.Give Curcumol,Curcumin,total saponins of Panax notoginseng of three groups of drug intervention,using MTT method to detect apoptosis and to choose the suitable concentration,The expression level of ROS(Reactive oxygen species,reactive oxygen)was detected by DCF method.The mRNA expression of inflammatory factors TNF-alpha,IL-6,IL-1 beta and fibrotic factor alpha-SMA,Smad2/3 and TGF-beta was detected by real time fluorescence quantitative PCR,and Western blot was used to detect the level of P-NF-kappa B and protein phosphorylation.ResuLts:1、After stimu Lating macrophages Raw264.7 with CSE,compared with the normal group,the fluorescence intensity of the model group increased and the expression level of ROS increased.Compared with the model group,the luminance of the drug group decreased in varying degrees,and the expression level of ROS decreased.The reduction of curcumol group was the most obvious,and P<0.05 was statistically significant.2、Curcumin,curcumol and total saponins of Panax could inhibit the NF-kappa B signaling pathway,down regulate the expression level of inflammatory cytokines TNF-alpha,IL-6 and IL-1 beta,and alleviate inflammation.3、After the inhibition of NF-kappa B with NF-kappa B inhibitor Bay 11-7082,the mRNA expression level of the inflammatory factor IL-6 and IL-1 beta in the curcumin group was obviously increased,and the P<0.05 was statistically significant.The mRNA expression level of TNF-alpha had a certain tendency to rise.the mRNA expression level of inflammatory factor TNF-alpha,IL-6 and IL-1 beta increased in total saponins of Panax group.The expression level of IL-6 and IL-1 mRNA in the curcumol group decreased,The level of mRNA expression of TNF-alpha has a tendency to rise.4、The expression level of Smad2/3 protein in curcumin group and curcumol group decreased,P<0.05,which was statistically significant.The level of mRNA expression of fibrotic factor alpha-SMA,Smad2/3 and TGF-beta was decreased.The expression level of Smad2/3 protein phosphorylation and the level of mRNA expression of Smad2/3 and TGFbeta in total saponins of Panax group were increased,and P<0.05 was statistically significant.the expression level of alpha-SMA mRNA in total saponins of Panax group was decreased,P<0.05 was statistically significant.5、After inhibiting the expression of NF-kappa B with NF-kappa B inhibitor Bay 11-7082,the expression level of alpha-SMA in the curcumin group increased,the expression level of Smad2/3 protein phosphorylation increased and P<0.05,and statistically significant compared with the model group.The expression level of mRNA in the curcumol group increased.The expression level of phosphorylation of Smad2/3 protein was significantly decreased,P<0.05 was statistically significant.the expression level of alpha-SMA mRNA in the total saponins of Panax group decreased slightly,and the expression level of Smad2/3 protein phosphorylation decreased,P<0.05 was statistically significant.Conclusion:1、Curcumol,curcumin and total saponins of Panax reduce the oxidative stress by reducing the release of ROS in macrophages.At the same time,by inhibiting the NF-kappa B signaling pathway,down regulating inflammatory cytokines TNF-alpha,IL-6,IL-1 beta expression level,alleviate inflammatory reaction.2、The main component of zedoary curcumae,curcumol and curcumin can reduce the level of mRNA expression of fibrotic factor alpha-SMA,Smad2/3 and TGF-beta by inhibiting the TGF-beta/Smads signaling pathway.