牙本质基质蛋白-1对白细胞介素-1β诱导的大鼠软骨细胞炎症损伤的研究

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Abstract

Objective To explore the biological effect of Dentin Matrix Protein-1 (DMP-1) on chondrocytes in osteoarthritis (OA).Methods The expression of DMP-1 in cartilage cells of OA rat model was analyzed with Affymetrix microarray.The chondrocyte was isolated，cultured and transfected with DMP-1 gene.Subsequently，the effect of DMP-1 on the proliferation of chondrocyte was examined with CCK-8 assay.ELISA assay was performed for detection of chondrocyte glycosaminoglycan (GAG) secretion and proinflammatory cytokines (IL-6 and TNF-α).The expression of Col2a1 was observed by immunofluorescence.RT-qPCR was to evaluate the expression levels of DMP-1，Sox9，Acan，and Col2a1 genes.Western Blot assay was conducted for DMP-1，Sox9，Acan，Col2a1 expressions.Flow cytometry and Calcein AM/PI staining assays were conducted for detection of the anti-apoptosis ability of chondrocyte treated with DMP-1.Results The Microarray analysis showed that the expression level of DMP-1 in OA group was lower than that in Control group in vivo (P＜0.05)；The expression level of DMP-1 in chondrocytes of OA group was also lower than that in Control group in vitro (P＜0.001).After transfection with DMP-1 gene，the expression level of DMP-1 in chondrocytes and chondrocytes induced by IL-1β was higher than that in the non-transfected group.The viability of chondrocytes induced by IL-1β was higher in transfected DMP-1 chondrocyte than that in the non-transfected group (P＜0.001).The expression of GAGs and immunofluorescence of Col2a1 in the transfected DMP-1 chondrocytes induced by IL-1β was significantly higher than that in the untransfected group，as well as the expression levels of Sox9，Acan and Col2a1 were significantly higher than that in the untransfected group (P＜0.001).The levels of IL-6 and TNF-α and apoptosis of transfected DMP-1 chondrocytes induced by IL-1β were significantly lower than those of non-transfected group (P＜0.001). Conclusion DMP-1 can be used to transfect chondrocytes and reduce the progression of OA.

Key words： dentin matrix protein-1 cartilage cells inflammation osteoarthritis

Corresponding Authors: Zhao Ping

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	Lu Bin
	Cheng Jiafeng
	Wang Yang
	Ji Lin
	Wang Yangsong
	Zhao Ping

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Lu Bin,Cheng Jiafeng,Wang Yang, et al. Experimental Study of Dentin Matrix Protein-1 in Interleukin-1β induced Rat Chondrocyte Inflammatory Lesion[J]. sygkzz, 2022, 28(11): 991-.

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http://www.sygkzz.com/EN/ OR http://www.sygkzz.com/EN/Y2022/V28/I11/991

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