The Expressions Of NF-κB In Asthma Rats And Modulation By Curcumin

OBJECTIVES:(1)To observe the protein expressions of NF-κB in rats asthma;(2)toinvestigate the relation of NF-κB and GSH,MDA and its influence on the formation ofairway inflammation;(3)to study the role of Curcumin in regulating the activity ofNF-κB.METHODS:36 male rats were randomly divided into the control group (groupA),asthma group(group B), Curcumin group(group C),each group have 12 rats.In theexperiment,the rat model of animals was established by the ovalbumin(OVA) challengemethods,but the control group were challenged by Saline instead of OVA.All the animalswere sacrificed after the last challenge.The lung tissue was gained from the leftlung,Bronchoalveolar lavage fluids(BALF) was gained from the right lung;The total cellnumbers, differentiaton cell numbers and the numbers of EOS were counted by differentcount fluids;The concentrations of MDA and GSH in serum and BALF were measuredrespectively by TAB and dinitrobencene methods. The concentrations of IL-5 in serum andBALF were measured by sandwich ELISA.the condition of inflammation cell infiltrationaround the bronchus smooth muscle proliferation were measured by light microscpe;theultrastructure of bronchus and pulmonary alveolusⅡtype epithelial cell and pulmonaryarterioles were observed by transmission electron microscope (TEM);the expressions ofNF-κBp65 and Eotaxin were determinded by immune -histochemistry techniques;the resultof immunhistochemistry were checked by the picture analyse meter,the obtained LD andthe concentration of IL-5were checked out by statistics analyse;the concentrations of MDAand GSH wre also checked out by statistics analyse.RESULTS:(1)The conditions of inflammation cell in BALF:The total cell numbers in BALF of group B[(5.64±1.12)×10~9/L] were significantly higher than that of group Aand group C [(2.58±0.22)×10~9/L,(3.49±0.97)×10~9/L, respectively](P＜0.01).The ratiosof esosinophil to the total cell numbers (EOS%),The ratios of neutrophil to the total cellnumbers(NEU%) and the ratios of lymphocytes to the total cell numbers(LYM%) of groupB were significantly higher than that of group A[(11.13±3.65)%, (15.37±4.13)%and(21.21±4.27)%] versus [(1.25±0.56)%,(11.02±3.02)%, (13.12±4.13),respectively](P＜0.01).The EOS% of group C (5.67±2.16)% wassignificantly lower than that of groupB(P＜0.01).The NEU% of group C[(13.46±1.69)%] was lower than that of group B (P＜0.05). (2) The observation results of pulmonary histopathology:HE-stained lung ofrats showed inflammation cells infiltraion around the bronchus and the vascula which weremainly lymphocytes, eosinophils and neutrocytesr, bronchus mucus bolting.Airwayepithelium damaging,basement membrane and smooth muscle thickening in group B.Respectively, there were different changes in group C,but the pathwaysologic alteration ofgroup C were much lighter than those of group B.The above-mentioned parameters werenot significant in group A.(3)The observation results of TEM:TEM showed normalpulmonary alveolus diaphragm structure,normal pulmonary alveolusⅡtype epithelial cellin group B,pulmonary alveolus diaphragm proliferation, lamellasome vacuolization,basemembrane prolifeation,inflam -mation cell infiltration,pulmonary alveolusⅡtypeepithelial cell degranulation in group A;compared with group A, the proliferation ofsmooth muscle cells and the constriction of artioles of Curcumin group were significantlylighter;(4) The concentrations of IL-5 in serum and BALF: the concentrations IL-5 inserum of group B[[26.61±2.84(ng/ml)]] was significantly higher than that of controlgroupA[1.85±0.56(ng/ml)],group C[9.56±1.61(ng/ml)] (P＜0.01,repectifily);theconcentration of IL-5 in BALF of group B[24.28±2.72(ng/ml)] was significantly higherthan that of control group[1.31±0.19(ng/ml)],group C[8..57±1..05(ng/ml)](P＜0.01,repectifily). (5) The concentrations of malondiadehyde (MDA) and reducedglutathione(GSH) in serum and BALF:the concentrations MDA in BALF of groupB[5.99±0.45(nmol/ml)] was significantly higher than that of control groupA[1.71±0.37(nmol/ml)]and group C[1.89±0.30(nmol/ml)](P＜0.01); theconcentrations MDA in serum of group B7.05±2.09(nmol/ml)]was significantly higherthan that of control groupA[2.64±0.70(nmol/ml)](P＜0.01)and groupC[3.50±1.01(nmol/ml)](P＜0.01, respectively);the concentrations GSH in serum of groupB[119.90±31.36(mg/L)] was significantly lower than that of control groupA[(283.84±68.20)mg/L] and group C[(184.30±36.29)mg/L](P＜0.01);the concentrationsGSH in BALF of group B[(87.37±16.86)mg/L] was significantly lower than that ofcontrol groupA[(231.20±25.81)mg/L] and group C[(138.99±59.43)mg/L](P＜0.01)(6)The results of Immunohistochemistry: Immunohistochemistry showed that theprotein expressions of the subunit P65 of NF-κB were not only in cytoplasm but also innucleus of those cells. The expressions of group B were significantly higher than those ofgroupA,groupC(P＜0.01, respectively);none expression almost was done in group A.Immunohistochemistry also showed that the protein of Eotaxin were mainly in cytoplasmof the bronchus epitheial cells.the nucleus were negative. The expressions of group B weresignificantly higher than those of group A,group C(P＜0.01, respectively);noneexpression almost was done in group A.(7) The results of correlation analysis:Therewas a significant correlation between the concentration of IL-5 in BALF,the content ofEotaxin,the content of NF-κBp65,the numbers of EOS in BALFand the concentration ofMDA in BALF respectively;There was a significant negative correlation between theconcentration of GSH in BALF and the content of Eotaxin,the content of NF-κBP65,thenumbers of EOS in BALF, respectively.CONCLUSIONS:(1) The subunit p65 of NF-κB are strongly expressed in asthma inrats, the epithelial cells and many other inflammation cells are the chief derivedcells.Eotaxin and IL-5 are also strongly expressed following with NF-κB.It is concludedthat the activity of signal pathways NF-κB is excess activating;(2) There was a significantpositive correlation between the content of NF-κBp65, Eotaxin, IL-5, and MDArespectively; but the correlation with GSH was negative.(3)Curcumin can significantlyhold down the activity of NF-κB, significantly hold down the airway inflammation cell infiltration of asthma.The inflammation medium significantly are reduced byCurcumin,through the inhibition of tranfer into nucleus and binding to DNA.This action ofCurcumin is mainly contributed to the activity of anti-oxidation.