Treatting Effect Of Huashi Xingyu Qingre Prescription On Oral Lichen Planus And Its Regulatory Effect On Notch/NF-κB Signaling Pathway

Oral lichen planus(OLP) is an oral mucousal disease of unknownpathogenesis.It has been recognised as a kind of precancerous condition byworld hygiene organization because of its tendency to cancer. There has notbeen effective clinical treatment for this disease. Huashi Xingyu Qingreprescription was found by Professor Xu Yanzhi according to the theory ofChinese traditional medicine and clinical study over many years. It wasdemonstrated to be an effective treatment after many years' preliminaryclinical observation. But the exact mechanism of Huashi Xingyu Qingreprescription is unknown and needs further research. There were142differentially expressed genes and8signaling pathways of OLP which hadbeen selected by microarray in our initial research. The nuclear factor-kappaB pathway is closely related to OLP. Recently, increasing evidence indicatedthat the interplay between NF-κB and Notch pathway was involved in manykinds of cancer. However, to our knowledge, there has been no report on therole of Notch/NF-κB signaling pathway in OLP. Therefore, we detected theNF-κB, Notch-1and Jagged-1expressions in oral lesions tissues byimmunohistochemistry methods, and discussed the mechanism of Notch-1/NF-κB signaling system in the development and progression of OLP;Meanwhile, we detected interleukin6(IL-6),vascular endothelial growthfactor (VEGF) and soluble cell adhesion molecules-1(sICAM-1) in serum andsaliva of patients with OLP by enzyme-linked immunosorbent assay, andobserved the changes of clinical lesions and the cell factor levels of patientsbefore and after treatment. Then we discussed the clinical effect andmechanism of Huashi Xingyu Qingre prescription, which may be establish abasis on clinical medication. Part One Expression and clinicopathological significance of NF-κB,Notch-1and Jagged-1in oral lichen planusObjective: This study was designed to investigate the expression ofNF-κB, Notch-1and Jagged-1in human oral squamous cell carcinoma, orallichen planus and normal oral mucosa tissue, and analyze their correlations.And then we discussed the role of Notch-1/NF-κB signaling system in thedevelopment and progression of oral lichen planus.Methods: Immunohistochemical staining was used to measure NF-κB,Notch-1and Jagged-1in45cases oral squamous cell carcinoma tissues,52cases oral lichen planus tissues and30cases normal mucosa tissues.And thedifferences of groups for immunohistochemical staining and the correlation ofthree proteins were analyzed by statistical methods.Results:1NF-κB which mostly expressed in cytoplasm, was expressed in oralsquamous cell carcinoma tissue, oral lichen planus tissue and normal mucosatissue. Its expression in oral lichen planus and oral squamous cell carcinomatissue was much stronger than that in normal mucosa tissues(Z=-7.709,P=0.000and Z=-6.605, P=0.000), and its expression in oral lichen planustissue was weaker than that in oral squamous cell carcinoma tissue (Z=-2.028,P=0.022).2Notch-1which mostly expressed in cytoplasm and caryon wasexpressed in oral squamous cell carcinoma tissue, oral lichen planus tissue andnormal mucosa tissue. Its expression in oral lichen planus and oral squamouscell carcinoma tissue was much stronger than that in normal mucosatissues(Z=-6.528, P=0.000and Z=-6.289, P=0.000), and its expression in orallichen planus tissue was weaker than that in oral squamous cell carcinomatissue (Z=-2.027, P=0.021).3Jagged-1which mostly expressed in cytoplasm and cytomembrane wasexpressed in oral squamous cell carcinoma tissue, oral lichen planus tissue andnormal mucosa tissue. Its expression in oral lichen planus and oral squamous cell carcinoma tissue was much stronger than that in normal mucosatissues(Z=-6.630, P=0.000and Z=-6.689, P=0.000), and its expression in orallichen planus tissue was weaker than that in oral squamous cell carcinomatissue (Z=-2.646, P=0.004).4In oral lichen planus tissue, NF-κB expression level had correlationwith Notch-1expression (r=0.333, P=0.016), NF-κB expression level hadcorrelation with Jagged-1expression (r=0.284, P=0.041), Notch-1andJagged-1expression has significant correlation (r=0.440, P=0.001); In oralsquamous cell carcinoma tissue, NF-κB expression level had correlation withNotch-1expression (r=0.695, P=0.000), and NF-κB expression level hadcorrelation with Jagged-1expression (r=0.330, P=0.027). Then Notch-1andJagged-1expression has significant correlation (r=0.443, P=0.002).Conclusion:1NF-κB, Notch-1and Jagged-1expression level in oral lichen planustissue and oral squamous cell carcinoma tissue were higher than that in normalmucosa tissue, which had statistical difference.2NF-κB, Notch-1and Jagged-1had the significant correlation in orallichen planus tissue and oral squamous cell carcinoma tissue, which showedthat the abnormal activition of Notch-1/NF-κB signaling pathway might takeeffects on the progress of oral lichen planus. Part Two Effect of Huashi Xingyu Qingre Prescription on expressionsof cell factors which related to Notch/NF-κB signalingpathway in oral lichen planusObjective: This research was designed to estah1ish thin layerchromatography (TLC) for the identification of Huashi Xingyu Qingreprescription to achieve its quality contro1, and evaluate its effect to treatpatients with Oral lichen planus (OLP),and then analysis of its mechanism. Methods: Cortex phellodendri and paeoniae rubra in Huashi XingyuQingre prescription were identified by TLC. The method of enzyme-linkedimmunosorbent assay (ELISA) was used to detect interleukin6(IL-6),vascular endothelial growth factor (VEGF) and soluble cell adhesionmolecules-1(sICAM-1) in serum and saliva of30patients with OLP beforeand after treatment.Results: The corresponding TLC spots of all the samples were able to bedetected in TLC identification. After8weeks' treatment for the patients,18cases had significantly effective,10cases were improved and2cases wereineffective. The total effect was93.3%.The IL-6, VEGF and sICAM-1levelsof serum and saliva were significantly lower,and there were significantdifferences (P<0.05).Conclusion:1TLC method is simple, accurate, specific, and available for qualitycontrol of the preparation.2Huashi Xingyu Qingre prescription for OLP shows an obvious clinicaleffect.3The expression of cell factors which related to Notch/NF-κB signalingpathway was low,after therapy of Huashi Xingyu Qingre prescription.Then wedemonstrat that Huashi Xingyu Qingre prescription may be treat OLP throughregulating Notch/NF-κB signaling pathway.