|
|
Influences of Paeoniflorin on LPS-Induced Apoptosis and Osteogenic Differentiation of Periodontal Ligament Stem Cells by Regulating LKB1-AMPK Signaling Pathway |
MEI Mei, et al |
The 940th Hospital of Joint Logistics Support Force of Chinese People's Liberation Army, Gansu Lanzhou 730050, China |
|
|
Abstract Objective: To investigate the influences of paeoniflorin (PF) on lipopolysaccharide (LPS)-induced apoptosis and osteogenic differentiation of periodontal ligament stem cells (PDLSCs) by regulating liver kinase B1 (LKB1)-adenylate-activated protein kinase (AMPK) signaling pathway. Methods: PDLSCs in logarithmic growth phase were collected and separated into control group, LPS group (10μg/mL), LPS+PF low concentration (PF-L, 10μg/mL LPS+5μmol/L PF) group, LPS+PF medium concentration (PF-M, 10μg/mL LPS+10μmol/L PF) group, LPS+PF high concentration (PF-H, 10μg/mL LPS+20μmol/L PF) group, and LPS+PF-H +LKB1 inhibitor (radicicol, 10μg/mL LPS+20μmol/L PF+10μmol/L radicicol) group. ELISA was used to detect the levels of inflammatory factors [interleukin (IL)-1β, tumor necrosis factor-α (TNF-α), IL-6] in cells; alizarin red staining was used to observe the changes of mineralized nodules; alkaline phosphatase (ALP) was used to detect the cell osteogenic differentiation; cell apoptosis was detected by flow cytometry; the expression levels of osteogenic genes (OPN, RUNX2 and OCN) were detected by qRT-PCR; Western blot was used to detect the expression levels of LKB1-AMPK pathway protein and apoptosis protein (caspase-1). Results: Compared with the control group, the mineralized nodules were reduced in the LPS group, the apoptosis rate, the expression of caspase-1, the contents of TNF-α, IL-1β and IL-6 increased obviously, the level of ALP, the expression of OPN, RUNX2 and OCN, and the phosphorylation levels of LKB1 and AMPK decreased obviously (P<0.05); compared with the LPS group, the mineralized nodules were reduced in the LPS+PF-L group, the LPS+PF-M group, and the LPS+PF-H group, the apoptosis rate, the expression of caspase-1, the contents of TNF-α, IL-1β and IL-6 decreased obviously, the level of ALP, the expression of OPN, RUNX2 and OCN, and the phosphorylation levels of LKB1 and AMPK increased obviously, all were dose-dependent (P<0.05); however, radicicol reversed the effects of PF-H on LPS-induced apoptosis and osteogenic differentiation of PDLSCs (P<0.05). Conclusion: PF inhibits LPS-induced apoptosis of PDLSCs and promotes osteogenic differentiation by up-regulating LKB1-AMPK signaling pathway.
|
|
|
|
|
[1] Duan Y,An W,Wu Y,et al.Tetramethylpyrazine reduces inflammation levels and the apoptosis of LPS stimulated human periodontal ligament cells via the downregulation of miR 302b[J].Int Mol Med,2020,45(6):1918-1926. [2] Guo L,Li L.LIN28A alleviates inflammation,oxidative stress,osteogenic differentiation and mineralization in lipopolysaccharide (LPS)-treated human periodontal ligament stem cells[J].Exp Ther Med,2022,23(6):411. [3] Zheng W,Wang S,Wang J,et al.Periodontitis promotes the proliferation and suppresses the differentiation potential of human periodontal ligament stem cells[J].Int Mol Med,2015,36(4):915-922. [4] Wen J,Xu B,Sun Y,et al.Paeoniflorin protects against intestinal ischemia/reperfusion by activating LKB1/AMPK and promoting autophagy[J].Pharmacol Res,2019,146:104308. [5] Li C,Xiao F,Wen Y,et al.Krüppel-like factor 5 -mediated Sirtuin6 promotes osteogenic differentiation and inhibits inflammatory injury of lipopolysaccharide-induced periodontal membrane stem cells by inhibiting nuclear factor kappa-B pathway[J].Bioengineered,2022,13(3):6966-6977. [6] Wang S,Jia D,Lu H,et al.Paeoniflorin improves myocardial injury via p38 MAPK/NF-KB p65 inhibition in lipopolysaccharide-induced mouse[J].Ann Transl Med,2021,9(18):1449. [7] Gurkan CG,Keles GC,et al.Histopathological and biochemical evaluation of paeoniflorin administration in an experimental periodontitis model[J].Oral Sci,2019,61(4):554-557. [8] 王如然,黄胜男,曹戬,等.miR-30a-5p通过靶向Runx2基因抑制成骨细胞分化[J].生物技术通讯,2020,31(2):142-147. [9] Zheng Y,Tao Y,Zhan X,et al.Nuclear receptor 4A1 (NR4A1) silencing protects hepatocyte against hypoxia-reperfusion injury in vitro by activating liver kinase B1 (LKB1)/AMP-activated protein kinase (AMPK) signaling[J].Bioengineered,2022,13(4):8349-8359. [10] Wang P,Ma T,Guo D,et al.Metformin induces osteoblastic differentiation of human induced pluripotent stem cell-derived mesenchymal stem cells[J].Tissue Eng Regen Med,2018,12(2):437-446. |
|
|
|