Effects of LPS-stimulated monocyte culture supernatant on the OPG/RANKL of osteoblastic cells

Abstract

Abstract: PURPOSE：To investigate the influence of Pg-LPS stimulated monocyte (RAW264.7) culture supernatant on the OPG/RANKL expression of osteoblastic cells (MC3T3-E1). METHODS：The culture supernatant of monocytes stimulated with Pg-LPS was applied to osteoblasts MC3T3-E1 with different diluted concentrations（10%, 20%, 30%, 40% and 50%） simultaneously for 24h, then RT-PCR was used to detect the expression changes of OPG/RANKL mRNA.Western blot was used to detect the expression changes of OPG/RANKL protein. The data was analyzed by ANOVA using SPSS 17.0 software package. RESULTS：After stimulation of different concentrations of inflammatory supernatant, the expressions of OPGmRNA and protein significantly decreased (P<0.05), whereas the expressions of RANKLmRNA and protein significantly increased(P<0.05), both of them were in a concentration-dependent manner. CONCLUSIONS：These results indicate that Pg-LPS stimulated RAW264.7 culture supernatant can inhibit the osteogenesis and differentiation of the osteoblasts through inhibiting the expression of OPGmRNA and protein of osteoblastic cells, while increasing the expression of RANKLmRNA and protein in a concentration-dependent manner .

Key words: LPS, Monocyte, Osteoblasts, OPG/RANKL Shanghai J Stomatol, 2015, 24(4):428-432.

CLC Number:

R781.4

WANG Hai-yan, QIN Hua-xiang, DENG Hui, SUN Chao-fan, HU Rong-dang. Effects of LPS-stimulated monocyte culture supernatant on the OPG/RANKL of osteoblastic cells[J]. Shanghai Journal of Stomatology, 2015, 24(4): 428-432.

References

[1] Van Gastel J, Quirynen M, Teughels W, et al. Longitudinal changes in microbiology and clinical periodontal variables after placement of fixed orthodontic appliances[J]. J Periodontol, 2008, 79(11): 2078-2086.
[2] Van Dyke TE. Control of inflammation and periodontitis[J]. Periodontology 2000, 2007, 45(1): 158-166.
[3] Czura CJ, Tracey KJ. Targeting high mobility group box 1 as a late-acting mediator of inflammation[J]. Crit Care Med,2003,31(1 Suppl): S46-50.
[4] Nason R, Lee DH, Jung J Y, et al. Radiographic and micro-computed tomographic imaging of lipopolysaccharide mediated bone resorting [J]. Ann Otol Rhinol Laryngol, 2009, 118(5): 391-396.
[5] 王海燕, 邓辉, 徐春燕, 等. 脂多糖刺激单核巨噬细胞培养上清液对成骨细胞成骨特性的影响[J]. 中华口腔正畸学杂志, 2011, 18(2): 92-96.
[6] Takahashi N, Udagawa N, Suda T. A new member of tumor necrosis factor ligand family, ODF/OPGL/TRANCE/RANKL, regulates osteoclast differentiation and function[J]. Biochem Biophys Res Commun, 1999, 256(3): 449-455.
[7] Pelt P, Zimmermann B, Ulbrich N, et al. Effects of lipopolysaccharide extracted from Prevotella intermedia on bone formation and on the release of osteolytic mediators by fetal mouse osteoblasts in vitro [J]. Arch Oral Biol, 2002, 47(12): 859-866.
[8] Kwak HB, Sun HM, Ha H, et al. Tanshinone IIA suppresses inflammatory bone loss by inhibiting the synthesis of prostaglandin E2 in osteoblasts[J]. Eur J Pharmacol,2008,601(l-3):30-37.
[9] Kawashima N, Stashenko P. Expression of bone-resorptive and regulatory eytokines in murine periapical inflammation[J]. Arch Oral Biol, 1999, 44(1): 55-66.
[10] Wang Y, Wang HY, Ye Q, et al. Co-regulation of LPS and tensile strain down regulating osteogenicity via c-fos expression[J]. Life Sci, 2013, 93(1): 38-43.
[11] Takahashi N, Udagawa N, Suda T. A new member of tumor necrosis factor ligand family, ODF/OPGL/TRANCE/RANKL, regulates osteoclast differentiation and function[J]. Biochem Biophys Res Commun, 1999, 256(3): 449-455.
[12] Liu D, Xu JK, Figliomeni L, et al. Expression of RANKL and OPG mRNA in periodontal disease: possible involvement in bone destruction [J]. Int J Mol Med, 2003, 11(1): 17-21.
[13] Horowitz MC, Xi Y, Wilson K. Control of osteoclastogenesis and bone resorption by members of the TNF family of receptors and ligands [J]. Cytokine Growth Factor Rev, 2001, 12(1): 9-18.
[14] Kim KW, Suh SJ, Lee TK, et al. Effect of safflower seeds supplementation on stimulation of the proliferation, differentiation and mineralization of osteoblastic MC3T3-E1 cells[J]. J Ethnopharmacol, 2008, 115(1): 42-49.

[1]	HU Xi, LUO Jun. Role and mechanism of miR-497-5p in the differentiation and mineralization of pre-osteoblast MC3T3-E1 [J]. Shanghai Journal of Stomatology, 2023, 32(1): 17-22.
[2]	PAN Qiao-ting, ZANG Xiao-long, SUN Zhao-wei, PAN Meng-qi, ZHU Xin-mei, LI Zhi-yong. The role of semaphorin 4D in the mechanism of bisphosphonate-related osteonecrosis of the jaw in rats [J]. Shanghai Journal of Stomatology, 2022, 31(6): 625-631.
[3]	GONG Xin-yi, HUANG Xiang-ru, ZHOU Si-ru, YANG Yi-ling, XU Hong-yuan, JIN An-ting, DAI Qing-gang, JIANG Ling-yong. Generation and validation of inducible osteoblast-specific Stat3 knockout mice [J]. Shanghai Journal of Stomatology, 2020, 29(4): 337-342.
[4]	MI Jing, YU Bo, YU Ya-qiong, LI Xiao-lin, DU Ya-xin, QIU Li-hong. Resveratrol counteracts lipopolisaccharide-mediated oxidative stress by upregulating SOCS-1 production [J]. Shanghai Journal of Stomatology, 2019, 28(3): 237-240.
[5]	ZHANG Man, LI Hao, XU Ting, LIU Jie, TAN Fei. Effect of strontium-containing cross-linked sodium alginate gel on adhesion and proliferation of pre-osteoblasts [J]. Shanghai Journal of Stomatology, 2019, 28(2): 123-127.
[6]	LI Xiao-lin, YU Ya-qiong, QIU Li-hong, GUO Jia-jie, SHAO Li-na, WANG Si-mo, YANG Di. Effects of Porphyromonas endodontalis lipopolysaccharides on the expression of monocyte chemotactic protein-1 in mouse osteoblasts [J]. Shanghai Journal of Stomatology, 2018, 27(1): 1-5.
[7]	ZHENG Jun-Yuan, HE Li, HU Tu-qiang.. The relationship between the effects of MTA on mRNA expression of four iconic proteins in cells of fetal rat skull and cell culture environment [J]. Shanghai Journal of Stomatology, 2016, 25(1): 16-21.
[8]	LI Wei-hong, HUANG Rui, LIN Hua, LI Qing-yan, ZHENG Xin-ying, LV Qiao, GAO Ning. Effect of beryllium on the morphology and chemical elements of cell membrane of Porphyromonas gingivalis [J]. Shanghai Journal of Stomatology, 2015, 24(4): 385-389.
[9]	LIN Zhi-kai, SHU Rong, SONG Zhong-chen, CHENG Lan, DONG Jia-chen, ZHANG Xiu-li. The effect of rhAm and EMPs on promoting differentiation of hBMSCs into osteoblasts [J]. Shanghai Journal of Stomatology, 2015, 24(4): 390-394.
[10]	LI Zhen-ling, ZHANG Zhi-min, LI Peng-lian, ZHANG Tong-fei, LI Tian-bo, LIU Fu-ping. Evaluation of the effect for ffh gene silencing on the aciduricity of fluoride resistant Streptococcus mutans [J]. Shanghai Journal of Stomatology, 2015, 24(4): 395-399.
[11]	PU Guang-rui, LIU Fa-yu, WANG Bo. Suppression of VEGF protein expression by arctigenin in oral squamous cell carcinoma [J]. Shanghai Journal of Stomatology, 2015, 24(4): 400-403.
[12]	HAO Yi, LI Jing, MA Jia-yin, LI Jun, XU Chun. Cylic stretching force-induced late apoptosis in human periodontal ligament cells [J]. Shanghai Journal of Stomatology, 2015, 24(4): 404-409.
[13]	HE Zhi-yan, CHENG Lan, WANG Yu-xia, HUANG Zheng-wei. Construction of luxS gene knockout mutant of Enterococcus faecalis [J]. Shanghai Journal of Stomatology, 2015, 24(4): 410-414.
[14]	XU Lin-yu, DONG Min, LU You-guang, LEI Lang. Study of factors affecting tooth discoloration during fixed orthodontics in vitro [J]. Shanghai Journal of Stomatology, 2015, 24(4): 415-418.
[15]	QIU Si-hui, WANG Jun. Effects of calcium hydroxide and Hoshino’s paste on the proliferation of porcine dental papilla cells [J]. Shanghai Journal of Stomatology, 2015, 24(4): 419-422.