| Backgorund Oral submucous Fibrosis(OSF) is a kind of chronic insidious disease and it predisposes to cancer. Histologically, OSF is characterized by epithelial atrophy and progressive accumulation of collagen fibers in the lamina propria and submucous of the oral mucosa with a progressive loss of vasularity. The majority of patients present with rigidity of lip, tongue, and palate leading to varying degrees of limitation of opening the mouth and tongue movement. A number of studies provide overwhelming evidence the betel quid is the main aetiological factor for OSF, OSF was reguarded as a precancerous condition by WHO, the malignant transformation rate of OSF was 1.7% in Hunan Province. The pathogenesis of OSF is still unknown, It may be associated with immunity, heredity, dysmetabolism and microcirculation disturbance. The complex pathogenesis of OSF makes it have no satisfying therapies. Formerly, surgery on the fibrous bands is not easy to be accepted by trauma and postoperative scar. Glucosida tripterygii TOTA and glucocorticoids also can not be taked for long-term by the side effect of gastralgia, osteoporosis, and hypocorticalism. Therefore, a new therapy for the treatment of OSF is necessary. According to the pathogenesis of OSF, it is efficient therapy by lowering the inflammatory reaction lesion and improving the microcirculation disturbance. As glucocorticoids are still the choice drug on fibrosis, and Salvia miltiorrhiza is good at activating blood circulation to dissipate blood stasis, moreover, Salvia miltiorrhiza can also inhibit osteoporosismade by glucocorticoids, our study like to evaluate the role of the combination of Salvia miltiorrhiza and low dose prednisolone on patients with oral submucous fibrosis, and explore the latent mechanism involved in the role of the combination of Salvia miltiorrhiza and low dose prednisolone on patients with oral submucous fibrosis in general level with proteomics technology.Object To establish tissue specimen bank and clinical information da-tabase of OSF. To explore the therapeutic effects of the combination of Salv-ia miltiorrhiza and low dose prednisolone on patients with oral submucous fibrosis and find the associated protein.Methods (1) To establish tissue specimen bank, and use Access 2003 and Excel 2003 to establish clinical information database of OSF.(2) There are 60 medium-term OSF patients and 60 advanced stage OSF patients, and each were randomly divided into the first groups (treated with both salvia miltiorrhiza and prednisolone) and the second group (treated separately with prednisolone). The therapeutic effects were compared among each group after the three month treatment.(3) Total proteins were extracted from OSF bucca tissues and the bucca tissues after treated by combination of Salvia miltiorrhiza and low dose prednisolone on patients with oral submucous fibrosis and were separated by immobilized PH gradient two-dimensional gel electrophoresis to establish the expression maps of proteomics. The differential expressed proteins between the two groups were analyzed with PDQuest image analysis software, and identified with MALDI-TOF-MS (matix-assisted laser desorption/ionization time of flight mass spectrometry).(4) Differentially expression level of selected protein was validate by Western blot and Immunohistochemistry analysis.Results (1) Tissue specimen bank and clinical information database of OSF were successfully constructed. More than 300 patient's clinical information was included in the information database which provides conveniences for subsequent research.(2) Both significant differences were found in the lesion area of the medium-term cases and the advanced stage cases of the first groups between prior treatment (10.37±3.40 cm2,19.60±3.27 cm2) and post treatment (5.90±4.10 cm2,16.33±4.02 cm2) (P<0.05); and also both significant differences were found in the mouth opening between prior treatment (3.41±0.77 cm,1.98±0.39 cm) and post treatment (3.87±0.67 cm,2.26±0.46 cm) (P<0.05). There were significant differences in the lesion area and mouth opening of the medium-term cases of the second groups between prior treatment (10.87±3.18 cm2,3.57±0.75 cm) and post treatment (6.70±3.75 cm2,3.97±0.69 cm) (P<0.05); but there was no significant difference in the lesion area and mouth opening of the advanced stage cases of the second groups (P>0.05). There was significant difference in the therapeutic efficacy between the first group (70%) and the second group (16.67%) of the advanced stage cases (P<0.05); but there was no significant difference in the clinical effects between the two groups of the medium-term cases (P>0.05). The side effect of prednisolone could be reduced while applied with salvia miltiorrhiza together.(3) Two-dimensional gel electrophoresis profiles with clear background, well reproducibility and high quality were obtained. Twenty-three specially expressed proteins were identified to be related to combination of Salvia miltiorrhiza and low dose prednisolone.4 spots (14-3-3σ,K4,Serotransferrin,Hemoglobin subunit beta) expressed highly and 19 spots (K6,α-SMA, MYL,TPM, ANXA2, S100A7, GAPDH, HspB5, et al) expressed lowly in therapy group specimens.(4) By analysis and classification of those proteins,14-3-3σprotein was considered to be the therapy on combination of Salvia miltiorrhiza and low dose prednisolone-associated protein candidate. Western blot exhibited an increase expression of 14-3-3σprotein. Immunohistochemical analysis of tissues showed that expression of 14-3-3σprotein in normal tissues and treated tissues were higher than in OSF tissues.Conclusion (1) Tissue specimen bank and clinical information database of OSF which were successfully constructed can be used for the study of OSF.(2) There is obvious advantage in treating OSF by the combination of Salvia miltiorrhiza and prednisolone.(3) 14-3-3σ,K4,Serotransferrin,Hemoglobin subunit beta,K6, a-SMA, MYL, TPM, ANXA2, S100A7, GAPDH and HspB5 could be thinked associated to the treatment of combination of Salvia miltiorrhiza and prednisolone on OSF.(4) 14-3-3σcould be used as indicator to predict the mechanism of combination of Salvia miltiorrhiza and prednisolone on OSF. |
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