Anti-SHH (aa 181-280) polyclonal antibody

Shh/Gli1 signaling plays important roles in development of spinal cord. How it is involved in spinal cord injury (SCI) remains unclear. In this study, we explored the roles of Shh/Gli1 signaling in SCI by using Shh signaling reporter Gli1(lz) mice and Gli1 mutant Gli1(lz/lz) mice. For detecting the Shh/Gli1 signaling after SCI, X-gal staining and double-immunostaining of Shh/PDGFR-beta, Shh/GFAP and LacZ/GFAP was conducted at 3 days post injury (dpi) on Gli1(lz) mice. To investigate the effects of Gil1 mutation on pathological changes after SCI, astrocytic proliferation and the content of intra-parenchymal Evans Blue were evaluated at 7dpi in wild-type and Gli1(lz/lz )mice. Furthermore, locomotor recovery was assessed by BMS scoring at 1, 3, 5 and 7dpi. The results of X-gal staining and immunohistochemistry showed that Shh/Gli1 signaling was mainly activated in reactive astrocytes after SCI. The 5-bromo-2-deoxyuridine (BrdU) incorporation assay showed that mutation of Gil1 did not affect the proliferation of astrocytes. However, the leakage of Evans Blue was significantly increased in the injured cord of Gli1(lz/lz )mice compared to wild-type mice. In addition, locomotor recovery was significantly impaired in the Gli1(lz/lz) mice. The findings demonstrated that Shh/Gli1 signaling could be induced in reactive astrocytes by SCI, and plays important role in permeability of blood-spinal cord barrier (BSCB) and locomotor recovery after SCI.

Background/Aims: To investigate the effect and the possible mechanism of lanthanum citrate on the proliferation and apoptosis of human hepatocellular carcinoma (HCC) cell line SMMC-7721 through the Hedgehog (Hh) signaling pathway. Materials and Methods: Different concentrations of lanthanum citrate and KAAD-cyclopamine (the Hh signaling pathway representative inhibitor) were used to treat SMMC-7721 cells. Cell proliferation was detected using Methylthiazolyldiphenyl-tetrazolium bromide (MTT) assays. Cell apoptosis was detected using flow cytometry analysis of Annexin V-FITC/propidium iodide (PI). The protein expressions of regulatory genes, such as cell cycle protein D1 (CyclinD1), cyclin-dependent kinase inhibitor 1 (p21), cysteinyl aspartate specific proteinase 3 (Caspase-3), B-cell lymphoma-2 (Bcl-2), glioma-associated oncogene homolog 1 (Gli1), and sonic hedgehog (Shh) were quantified using Western blot assays. The mRNA expressions of Gli1 and Shh were tested using quantitative real-time polymerase chain reaction (qRT-PCR) assays and the protein expressions of Gli1 and Shh were determined using immunofluorescence assays. Results: The Annexin V-FITC and PI double staining results revealed that the 0.1 mM lanthanum citrate group and the 15 mu M KAAD-cyclopamine group had both increased the apoptosis rate of SMMC-7721 cells. Both lanthanum citrate and KAAD-cyclopamine downregulated the protein expressions of CyclinD1, Bcl-2, Gli1, and Shh and upregulated the protein expressions of p21 and Caspase-3. Additionally, the immunofluorescence results revealed that the protein expressions of Gli1 and Shh were significantly decreased in both the lanthanum citrate group and the KAAD-cyclopamine group compared to the control group. Conclusion: Lanthanum citrate inhibits proliferation and promotes apoptosis in HCC SMMC-7721 cells by suppressing the Hh signaling pathway.