O papel de proteínas das vias WNT/ß-Catenina, TGF-ß/BMP4 e SHH na odontogênese e odontomas / The role of proteins from the WNT/ß-Catenin, TGF-ß/BMP4 and SHH pathways in odontogenesis and odontomas

O desenvolvimento do dente depende de uma série de interações sinalizadoras recíprocas entre o epitélio oral (EO) e o ectomesênquima derivado da crista neural, a via WNT com o TGF-ß e BMP4 tem sido implicada na tumorigênese. A via de sinalização tipo Wingless (Wnt) / ß-catenina é essencial para a ativação precoce da odontogênese e no desenvolvimento de tumores odontogênicos. O TGF-ß e as BMPs tem sido associadas aos processos de dentinogênese reacionária e reparadora. A sinalização de Shh pode regular a proliferação celular no ectomesênquima dentário, controlando assim a morfogênese dentária. O objetivo da pesquisa foi investigar a atuação de algumas proteínas das vias na odontogênese e na formação de odontomas e tumores odontogênicos mistos benignos, para isto, foi desenvolvido um estudo seccional restrospectivo e imuno-histoquímico contendo 23 odontomas compostos, 21 odontomas complexos, 17 germes dentários, 05 fibro-odontomas ameloblásticos e 01 fibroma ameloblástico. Os resultados encontrados demonstraram maiores imunoexpressões da via WNT/ß-catenina no epitélio dos germes dentários (p<0,001) e no fibroma ameloblástico, enquanto que, esteve no ectomesênquima dos odontomas (p<0,001) e fibro-odontomas ameloblásticos. A via WNT/ßcatenina correlacionou-se moderadamente e significativamente com a CK14 no epitélio (p = 0,007) dos odontomas. A BMP4 foi imunoexpressa, especialmente, no ectomesênquima dos odontomas complexos (mediana = 33,7; p<0,001). A via Shh foi mais imunoexpressa no epitélio dos germes dentários (p<0,001) e no ectomesênquima dos odontomas complexos (p=0,029). De forma similar, o TGFß apresentou maior imunoexpressão no epitélio dos germes dentários (p<0,001) e no ectomesênquima dos odontomas complexos (p = 0,002). O dente em desenvolvimento exibiu maiores concentrações para estas proteínas no epitélio odontogênico nas fases de botão e capuz e a expressão diferencial ocorreu, principalmente, no ectomesênquima dos tumores, o que indica que esse componente é de fato mais proliferativo (AU).

Tooth development depends on a series of reciprocal signaling interactions between oral epithelium (EO) and neural crest-derived ectomesenchyme, the WNT pathway with TGF-ß and BMP4 has been implicated in tumorigenesis. The Wingless (Wnt)/ß-catenin signaling pathway is essential for the early activation of odontogenesis and the development of odontogenic tumors. TGF-ß and BMPs have been associated with reactionary and reparative dentinogenesis processes. Shh signaling can regulate cell proliferation in dental ectomesenchyme, thus controlling dental morphogenesis. The objective of the research was to investigate the role of some proteins in the pathways in odontogenesis and in the formation of odontomas and benign mixed odontogenic tumors. tooth germs, 05 ameloblastic fibro-odontomas and 01 ameloblastic fibroma. The results found showed higher immunoexpressions of the WNT/ß-catenin pathway in the epithelium of tooth germs (p<0.001) and in ameloblastic fibroma, while it was in the ectomesenchyme of odontomas (p<0.001) and ameloblastic fibroodontomas. The WNT/ß-catenin pathway correlated moderately and significantly with CK14 in the epithelium (p = 0.007) of odontomas. BMP4 was immunoexpressed, especially in the ectomesenchyme of complex odontomas (median = 33.7; p<0.001). The Shh pathway was more immunoexpressed in the epithelium of tooth germs (p<0.001) and in the ectomesenchyme of complex odontomas (p=0.029). Similarly, TGF-ß showed higher immunoexpression in the epithelium of tooth germs (p<0.001) and in the ectomesenchyme of complex odontomas (p = 0.002). The developing tooth exhibited higher concentrations of these proteins in the odontogenic epithelium in the bud and cap phases and the differential expression occurred mainly in the ectomesenchyme of the tumors, which indicates that this component is in fact more proliferative (AU).

Effect and potential signaling pathway of hedgehog protein on the function of human retinal microvascular endothelial cell / 中华实验眼科杂志

Objective@#To explore the impact of Hedgehog protein on human retinal microvascular endothelial cell(HRMEC)and its signaling pathway.@*Methods@#The cultured HRMECs were divided into normal control group, 0.5 μmol/L agonist group and 1.0 μmol/L agonist group, and were cultured in medium with final concentration of 0, 0.5 and 1.0 μmol/L Hedgehog agonist, respectively; HRMECs cultured in high glucose medium were divided into high glucose control group, 1.5 μmol/L inhibitor group and 2.5 μmol/L inhibitor group.Erismodegib, the Smoothed inhibitor with final concentration of 0, 1.5 and 2.5 μmol/L was added into corresponding group, respectively.MTS method and Transwell cell migration method were used to detect the proliferation(A490 value)and relative mobility of HRMEC.The phosphorylation of PLCγ1, Akt and Erk proteins were detected by Western blot.@*Results@#The relative expression of Hedgehog protein in the high glucose control group was 6.24±0.11, which was significantly higher than 1.00±0.00 in the normal control group(t=667.573, P<0.001). The A490 value was 1.349±0.050 and 1.422±0.053, and the relative mobility rate was 2.34±0.14 and 3.59±0.32 in the0.5 μmol/L agonist group and the 1.0 μmol/L agonist group, respectively, which were significantly higher than 1.203±0.101 and 1.00±0.00 in the normal control group(all at P<0.01). The A490 value was 0.849±0.010 and 0.737±0.030, and the relative mobility rate was 0.43±0.02 and 0.27 ±0.01 in the 1.5 μmol/L inhibitor group and the 2.5 μmol/L inhibitor group, respectively, which were significantly lower than 1.000±0.040 and 1.00±0.00 in the high glucose control group(all at P<0.01). The phosphorylation ratios of PLCγ1, Akt and Erk in the 0.5 μmol/L agonist group and the 1.0 μmol/L agonist group were significantly higher than those in the normal control group(all at P<0.01). The phosphorylation ratios of PLCγ1, Akt and Erk in the 1.5 μmol/L inhibitor group and the 2.5 μmol/L inhibitor group were significantly lower than those in the high glucose control group(all at P<0.01).@*Conclusions@#High glucose induces the expression of Hedgehog protein in HRMEC.Hedgehog protein may regulate the function of HRMEC by regulating the phosphorylation of PLCγ1, Akt and Erk in G Protein-coupled receptors pathway.

Effect and potential signaling pathway of hedgehog protein on the function of human retinal microvascular endothelial cell / 中华实验眼科杂志

Objective To explore the impact of Hedgehog protein on human retinal microvascular endothelial cell(HRMEC) and its signaling pathway. Methods The cultured HRMECs were divided into normal control group,0. 5μmol/L agonist group and 1. 0μmol/L agonist group,and were cultured in medium with final concentration of 0,0. 5 and 1. 0μmol/L Hedgehog agonist,respectively;HRMECs cultured in high glucose medium were divided into high glucose control group,1. 5μmol/L inhibitor group and 2. 5μmol/L inhibitor group. Erismodegib,the Smoothed inhibitor with final concentration of 0,1. 5 and 2. 5 μmol/L was added into corresponding group,respectively. MTS method and Transwell cell migration method were used to detect the proliferation( A490 value) and relative mobility of HRMEC. The phosphorylation of PLCγ1, Akt and Erk proteins were detected by Western blot. Results The relative expression of Hedgehog protein in the high glucose control group was 6. 24±0. 11,which was significantly higher than 1. 00±0. 00 in the normal control group(t=667. 573,P<0. 001). The A490 value was 1. 349±0. 050 and 1. 422±0. 053,and the relative mobility rate was 2. 34±0. 14 and 3. 59±0. 32 in the 0. 5μmol/L agonist group and the 1. 0μmol/L agonist group, respectively, which were significantly higher than 1. 203 ± 0. 101 and 1. 00 ± 0. 00 in the normal control group(all at P<0. 01). The A490 value was 0. 849±0. 010 and 0. 737±0. 030,and the relative mobility rate was 0. 43 ± 0. 02 and 0. 27 ± 0. 01 in the 1. 5 μmol/L inhibitor group and the 2. 5 μmol/L inhibitor group, respectively,which were significantly lower than 1. 000±0. 040 and 1. 00±0. 00 in the high glucose control group(all at P<0. 01). The phosphorylation ratios of PLCγ1,Akt and Erk in the 0. 5μmol/L agonist group and the 1. 0μmol/L agonist group were significantly higher than those in the normal control group ( all at P<0. 01 ) . The phosphorylation ratios of PLCγ1,Akt and Erk in the 1. 5μmol/L inhibitor group and the 2. 5μmol/L inhibitor group were significantly lower than those in the high glucose control group ( all at P<0. 01 ) . Conclusions High glucose induces the expression of Hedgehog protein in HRMEC. Hedgehog protein may regulate the function of HRMEC by regulating the phosphorylation of PLCγ1,Akt and Erk in G Protein-coupled receptors pathway.

Molecular basis of basal cell carcinoma

Abstract: Basal cell carcinoma is the most common cancer, presenting low mortality but high morbidity, and it has as risk factor exposure to sunlight, especially UVB spectrum. The most important constitutional risk factors for basal cell carcinoma development are clear phototypes (I and II, Fitzpatrick classification), family history of basal cell carcinoma (30-60%), freckles in childhood, eyes and light hair. The environmental risk factor better established is exposure to ultraviolet radiation. However, different solar exposure scenarios probably are independent risk factors for certain clinical and histological types, topographies and prognosis of this tumor, and focus of controversy among researchers. Studies confirm that changes in cellular genes Hedgehog signaling pathway are associated with the development of basal cell carcinoma. The cellular Hedgehog signaling pathway is activated in organogenesis, but is altered in various types of tumors.

Role of Shh-PARP-1 signaling pathway in the protective effects of tea polyphenols against fatty acid-induced injury to islet microvessel endothelial function / 第二军医大学学报

Objective To investigate the role of Shh-PARP-1 signaling pathway in the protective effects of tea polyphenols against the fatty acid-induced islet microvessel endothelial function injury. Methods Mouse islet microvessel endothelial MS-1 cells were used in this study, and the cells were divided into normal control group, solvent group, fatty acid group (0. 25 mmol/L palmitic acid + 0. 5 mmol/L oleic acid), tea polyphenols group (25 μmol/L), fatty acid + tea polyphenols group, PARP-1 inhibitor (8 μmol/L BYK204165) + fatty acid group, PARP-1 inhibitor + fatty acid + tea polyphenols group, Shh inhibitor (2. 5 μmol/L cyclopamine) + fatty acid group, Shh inhibitor + fatty acid + tea polyphenols group and inhibitors of Shh and PARP-1 + fatty acid +tea polyphenols group. The changes of cell viability, apoptosis, nitric oxide (NO) synthesis and oxidative stress related indicators were examined in each group. Results After fatty acid treatment, the survival rate ofMS-1 cells was decreased, and the level of apoptosis was significantly increased (P0. 05). Conclusion Fatty acid can directly trigger islet microvessel endothelial function injury, and tea polyphenols shows a protective effect against the toxicity of fatty acid, which can be enhanced by inhibiting Shh-PARP-1 signal pathway.

Strontium ranelate promotes osteogenic differentiation of rat bone mesen-chymal stem cells through Hedgehog/Gli1 signaling pathway / 中国病理生理杂志

AIM: To explore whether strontium ranelate ( Sr ) promotes osteogenic differentiation of rat bone mesenchymal stem cells (BMSCs) through the Hedgehog/Gli1 signaling pathway.METHODS:BMSCs were isolated from 4-week-old rats by adherent culture and induced to differentiate into osteoblasts .According to the experimental purposes , the cells were exposed to different concentrations of Sr , cyclopamine ( Cy, an inhibitor of Hedgehog receptor ) or Gli1-siR-NA.The expression of Gli1 and Runx2 in the cells was detected by Western blotting .The activity of alkaline phosphatase ( ALP) was measured by the method of colorimetry , and the mineralized nodules were observed under microscope with aliz-arin red staining .RESULTS:Exposure to Sr at concentrations of 0.1 to 5 mmol/L for 7 d markedly increased the expres-sion of Gli1 in the BMSCs , and the increase in Gli1 expression was the most obvious following Sr exposure at concentration of 3 mmol/L.Cy at concentration of 10 μmol/L inhibited Sr-induced up-regulation of Gli1 expression.Transfection of the BMSCs with Gli1-siRNA not only obviously inhibited Sr-induced up-regulation of Gli1 and Runx2 ( a downstream protein of Gli1) expression, but also antagonized Sr-induced enhancement of ALP activity and the formation of mineralized nodules . CONCLUSION:The Hedgehog/Gli1 pathway is involved in Sr-induced osteogenic differentiation of rat BMSCs .