FBLN5 was regulated by PRDM9, and promoted senescence and osteogenic differentiation of human periodontal ligament stem cells.
作者: Mengyao Zhao ; Rong Rong ; Chen Zhang ; Haoqing Yang ; Xiao Han ; Zhipeng Fan ; Ying Zheng ; Jianpeng Zhang
Periodontal ligament stem cells (PDLSCs) are ideal seed cells for periodontal tissue regeneration. Our previous studies have indicated that the histone methyltransferase PRDM9 plays an important role in human periodontal ligament stem cells (hPDLSCs). Whether FBLN5, which is a downstream gene of PRDM9, also has a potential impact on hPDLSCs is still unclear.
Senescence was assessed using β-galactosidase and Enzyme-linked immunosorbent assay (ELISA). Osteogenic differentiation potential of hPDLSCs was measured through Alkaline phosphatase (ALP) activity assay and Alizarin red detection, while gene expression levels were evaluated using western blot and RT-qPCR analysis.
FBLN5 overexpression promoted the osteogenic differentiation and senescence of hPDLSCs. FBLN5 knockdown inhibited the osteogenic differentiation and senescence of hPDLSCs. Knockdown of PRDM9 decreased the expression of FBLN5 in hPDLSCs and inhibited senescence of hPDLSCs. Additionally, both FBLN5 and PRDM9 promoted the expression of phosphorylated p38 MAPK, Erk1/2 and JNK. The p38 MAPK pathway inhibitor SB203580 and the Erk1/2 pathway inhibitor PD98059 have the same effects on inhibiting the osteogenic differentiation and senescence of hPDLSCs. The JNK pathway inhibitor SP600125 reduced the senescence of hPDLSCs.
FBLN5 promoted senescence and osteogenic differentiation of hPDLSCs via activation of the MAPK signaling pathway. FBLN5 was positively targeted by PRDM9, which also activated the MAPK signaling pathway.
2023-08-21·International journal of molecular sciences
Inhibitory Effects of Simvastatin on IL-33-Induced MCP-1 via the Suppression of the JNK Pathway in Human Vascular Endothelial Cells.
An alarmin, interleukin (IL)-33 is a danger signal that causes inflammation, inducing chemotactic proteins such as monocyte chemoattractant protein (MCP)-1 in various cells. As statins have pleiotropic actions including anti-inflammatory properties, we investigated the effects of simvastatin on IL-33-induced MCP-1 expression in human umbilical vein endothelial cells (HUVECs). HUVECs were stimulated with IL-33 in the presence or absence of simvastatin. Gene expression and protein secretion of MCP-1, phosphorylation of mitogen-activated protein kinase (MAPK), nuclear translocation of phosphorylated c-Jun, and human monocyte migration were investigated. Immunocytochemical staining and Western immunoblot analysis revealed that IL-33 augmented MCP-1 protein expression in HUVECs. Real-time reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) showed that IL-33 significantly increased MCP-1 mRNA and protein secretion, which were suppressed by c-jun N-terminal kinase (JNK) inhibitor SP600125 and p38 MAPK inhibitor SB203580. Simvastatin inhibited IL-33-induced MCP-1 mRNA, protein secretion, phosphorylation of JNK and c-Jun. Additionally, the IL-33-induced nuclear translocation of phosphorylated c-Jun and THP-1 monocyte migration were also blocked by simvastatin. This study demonstrated that IL-33 induces MCP-1 expression via the JNK and p38 MAPK pathways in HUVECs, and that simvastatin inhibits MCP-1 production by selectively suppressing JNK. Simvastatin may inhibit the progression of IL-33-induced inflammation via suppressing JNK to prevent MCP-1 production.
2023-08-14·British journal of pharmacology
Inhibition of p38MAPK signal pathway alleviates radiation-induced testicular damage through improving spermatogenesis.
作者: Juan Yang ; Xiangying Ou ; Manling Shu ; Jie Wang ; Xuan Zhang ; Zhenyu Wu ; Wei Hao ; Huihong Zeng ; Lijian Shao
BACKGROUND AND PURPOSE:
How to prevent the damage of ionizing radiation to testis has become an urgent problem to be solved. The present aim is to investigate whether inhibition of p38MAPK signaling can alleviate radiation-induced testicular damage.
HE staining was used to measure the morphological changes of epididymis and testis after irradiation. Immunohistochemistry staining was used to assess the expression of PLZF, SOX9, p-p38MAPK in testis. Immunofluorescent staining was used to measure the expression of DDX4 and SCP3 in testis. RNA-Seq was used to profile gene expression in testis after irradiation. The expression of Mapk14, Atf2, Ddit3 and Ap1m1 genes was detected by qPCR. Western blotting was used to detect the protein expression of p38MAPK and p-p38MAPK in testis.
There was a dose-response relationship between testicular injury and ionizing radiation. The sperm quality was significantly decreased at 6 and 8 weeks after 6Gy of x-ray radiation. Radiation led to the decrease of PLZF+ cells and increase of SOX9+ cells in testis. There were 969 differential expressing genes in irradiated testis when compared to non-irradiated ones through RNA-Seq. The expression of genes related to p38MAPK signal pathway enriched by GO analysis was significantly increased in irradiated testis, which were further confirmed by qPCR. The protein expression of p-p38MAPK in testis was significantly increased at 28 days after irradiation. SB203580 treatment increased numbers of spermatozoa, the area and diameter of seminiferous tubules and numbers of SOX9+ cells in irradiated mice, which were consistent with the increased sperm movement rate and density under radiation with SB203580 administration. Furthermore, SB203580 treatment significantly increased numbers of SCP3+ cells in testis after irradiation, accelerating the process of spermatogenesis.
CONCLUSIONS AND IMPLICATIONS:
Ionizing radiation significantly changes testicular gene expression, in which p38MAPK signaling pathway was activated. Inhibition of p38MAPK signaling by SB203580 partially alleviated the testicular damage caused by radiation and accelerated the recovery of sperms through promoting spermatogenesis.