Chemopreventive effect of modified zeng-sheng-ping on oral squamous cell carcinoma by regulating tumor associated macrophages through targeting tnf alpha induced protein 6.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is the most common malignancy of the head and neck. Zeng-Sheng-Ping, composed of Sophora tonkinensis Gagnep., Bistorta officinalis Delarbre, Sonchus arvensis L., Prunella vulgaris L., Dioscorea bulbifera L., and Dictamnus dasycarpus Turcz., was regarded as an anti-cancer drug with significant clinical efficacy, but was discontinued due to liver toxicity. Our research group developed a modified Zeng-Sheng-Ping (ZSP-M) based on original Zeng-Sheng-Ping that exhibited high efficiency and low toxicity in preliminary investigations, although its pharmacodynamic mechanism is still unclear. Here, we aimed to elucidate the pharmacodynamic material basis of ZSP-M and investigate its chemopreventive effect on OSCC by modulating tumor associated macrophages (TAMs). METHODS: Components of ZSP-M were characterized using ultra-performance liquid chromatography-mass spectrometry. Chemopreventive effect induced by ZSP-M against experimental oral cancer was investigated using the 4-nitroquinoline N-oxide precancerous lesion mouse model. RNA sequencing analysis was used to gain a global transcriptional view of the effect of ZSP-M treatment. A cell co-culture model was used to study the targeted effect of ZSP-M on TAMs and the biological properties of OSCC cells and to detect changes in TAM phenotypes. The binding of ZSP-M active compounds to TNF alpha induced protein 6 (TNFAIP6) protein was analyzed by molecular docking and dynamic simulation. RESULTS: Forty main components of ZSP-M were identified, the most abundant of which were flavonoids. ZSP-M inhibited the degree of epithelial dysplasia in precancerous lesions by inhibiting the expression of the TNFAIP6 and CD163 proteins in the precancerous lesions of the tongue. ZSP-M inhibited proliferation, colony formation, migration and invasion of SCC7 cells by targeting TAMs. ZSP-M reduced the expression of CD163+ cells, inhibited the expression of TNFAIP6 protein, Arg1 mRNA and Il10 mRNA in TAMs, and reduced IL-10 cytokine release in the co-culture environment. This effect was maintained after the addition of recombinant TNFAIP6 protein. Computer simulations showed that trifolirhizin and maackiain are well-connected to TNFAIP6. CONCLUSIONS: ZSP-M counteracts the immunosuppressive action of TAMs by specific targeting of TNFAIP6, thereby exerting chemopreventive activity of OSCC.

Force-Induced Nitric Oxide Promotes Osteogenic Activity during Orthodontic Tooth Movement in Mice.

Objectives: The aim of this study was to investigate the effect of nitric oxide (NO) on orthodontic tooth movement and the regulatory effect on bone formation. Design: A mouse orthodontic tooth movement model was established to measure the level of releasing NO. Besides, orthodontic tooth movement distance and the bone formation in the tension side of the orthodontic tooth were also analyzed. In vitro, human periodontal ligament stem cells (hPDLSCs) were cultured under tensile force stimulation. The production of NO and the expression level of nitric oxide synthase (NOS) were detected after mechanical stimulation. Furthermore, the downstream cellular signaling pathway regulated by NO was also explored. Results: The generation of NO steadily increased throughout the orthodontic tooth movement in mice. Orthodontic tooth movement was decreased in the NOS inhibitor group while it was accelerated in the NO precursor group. Force-induced NO promoted the osteogenic differentiation of human hPDLSCs under tensile force stimulation. And force-induced NO in hPDLSCs regulated the PI3K/Akt/ß-catenin signal pathway. Conclusion: NO is involved in the regulation of orthodontic tooth movement and promotes bone formation on the tension side of the orthodontic tooth. The PI3K/Akt/ß-catenin pathway is one of the downstream cell signal transduction pathways of NO in the orthodontic process.

MicroRNA-21 promotes orthodontic tooth movement by modulating the RANKL/OPG balance in T cells.

OBJECTIVES: The present study was designed to investigate the effects of microRNA-21 (miR-21) on orthodontic tooth movement. METHODS: The orthodontic tooth movement model was established in C57BL/6 and miR-21-/- mice with or without implantation of activated T cells. Histological and histomorphometrical analyses were performed by hematoxylin-eosin staining. Tartrate-resistant acid phosphate staining was used to analyze the osteoclast numbers during tooth movement. Enzyme-linked immunosorbent assay, reverse transcription polymerase chain reaction, and immunohistochemistry analysis were used to examine the expression of RANKL and OPG. RESULTS: In miR-21-/- mice, the distance of tooth movement was retarded, the osteoclast number was decreased, and serum RANKL expression was strongly reduced. MiR-21 promoted the secretion of RANKL from activated T cells. Furthermore, activated T cells could partially rescue the decreased orthodontic tooth movement distance in miR-21-/- mice. MiR-21 was shown to promote orthodontic tooth movement by modulating the RANKL/OPG balance in T cells. CONCLUSIONS: The impact of miR-21 on tooth movement was better elucidated, furthering our understanding of its role and clinical applications in orthodontics.

D-Mannose Enhanced Immunomodulation of Periodontal Ligament Stem Cells via Inhibiting IL-6 Secretion.

Periodontal ligament stem cell- (PDLSC-) mediated periodontal tissue regeneration has recently been proposed for the new therapeutic method to regenerate lost alveolar bone and periodontal ligament. It was reported that both autogenic and allogeneic PDLSCs could reconstruct damaged periodontal tissues but the regeneration effects were not consistent. The effective methods to improve the properties of PDLSCs should be further considered. In this study, we investigated if D-mannose could affect the immunomodulatory properties of hPDLSCs. After being pretreated with D-mannose, hPDLSCs could inhibit T cell proliferation and affect T cell differentiation into Treg cells. We found that less IL-6 could be detected in D-mannose-pretreated hPDLSCs. In the D-mannose pretreatment group, induced Treg cell number would decrease if increased IL-6 levels could be detected. Our data uncovered a previously unrecognized function of D-mannose to regulate the immunomodulatory function of PDLSCs and that IL-6 might play a key role in this process. The results provided a property method to improve PDLSC-based periodontal regeneration.