Analysis of Effects of PTEN-Mediated TGF-β/Smad2 Pathway on Osteogenic Differentiation in Osteoporotic Tibial Fracture Rats and Bone Marrow Mesenchymal Stem Cell under Tension

IF 2.6 2区生物学 Q3 CELL BIOLOGY

Cellular Microbiology Pub Date : 2022-04-29 DOI:10.1155/2022/1004203

Shiyong Ling, Chen Yan, Kai Huang, Bo Lv, Hua Wang, Xiaoyan Wang, Jun Chen, Jingchuan Sun

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引用次数: 0

Abstract

Purpose. To discuss effects of phosphatase and tensin homolog protein (PTEN)-mediated transforming growth factor-β (TGF-β)/Smad homologue 2 (Smad2) pathway on osteogenic differentiation in osteoporotic (OP) tibial fracture rats and bone marrow mesenchymal stem cell (BMSC) under tension. Methods. A tibial fracture model was established. The rats were divided into sham-operated group and model group, and tibia tissue was collected. Purchase well-grown cultured rat BMSC, and use the Flexercell in vitro cell mechanics loading device to apply tension. The expression of PTEN was detected by qRT-PCR. After the BMSCs were transfected with si-PTEN and oe-PTEN, the force was applied to detect cell differentiation. The expression of TGF-β/Smad2 protein was detected by Western blot. The formation of calcium nodules in BMSC was detected by alkaline phosphatase (ALP) staining and alizarin red (AR) staining. Results. The expression of PTEN was higher in the model group and tension MSC group, and the expression of TGF-β and Smad2 protein was lower. The expression of TGF-β and Smad2 protein in oe-PTEN group was lower than the oe-NC group and control group. The expression of TGF-β and Smad2 protein in si-PTEN group was higher than the si-NC group and control group. The results of ALP staining and AR staining also confirmed the above results. Conclusion. PTEN-mediated TGF-β/Smad2 pathway may play a key role in the osteogenic differentiation of OP tibial fracture rats. Downregulation of PTEN and upregulation of TGF-β/Smad2 signal can promote the osteogenic differentiation of BMSC under tension.

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PTEN介导的TGF-β/Smad2通路对骨质疏松性胫骨骨折大鼠成骨分化和骨髓间充质干细胞张力作用的分析

目的。探讨磷酸酶和张力素同源蛋白(PTEN)介导的转化生长因子-β (TGF-β)/Smad同源物2 (Smad2)通路对张力作用下骨质疏松性(OP)胫骨骨折大鼠成骨分化及骨髓间充质干细胞(BMSC)的影响。方法。建立胫骨骨折模型。将大鼠分为假手术组和模型组，取胫骨组织。购买培养良好的大鼠骨髓间充质干细胞，使用Flexercell体外细胞力学加载装置施加张力。采用qRT-PCR检测PTEN的表达。si-PTEN和oe-PTEN转染骨髓间充质干细胞后，施加力检测细胞分化。Western blot检测TGF-β/Smad2蛋白的表达。碱性磷酸酶(ALP)染色和茜素红(AR)染色检测骨髓间充质干细胞钙结节的形成。结果。模型组和张力间充质干细胞组PTEN表达升高，TGF-β和Smad2蛋白表达降低。TGF-β和Smad2蛋白在e- pten组的表达低于e- nc组和对照组。si-PTEN组TGF-β和Smad2蛋白表达高于si-NC组和对照组。ALP染色和AR染色结果也证实了上述结果。结论。pten介导的TGF-β/Smad2通路可能在OP胫骨骨折大鼠成骨分化中起关键作用。下调PTEN和上调TGF-β/Smad2信号可促进紧张状态下BMSC的成骨分化。

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来源期刊

Cellular Microbiology 生物-微生物学

CiteScore

9.70

自引率

0.00%

发文量

审稿时长

3 months

期刊介绍： Cellular Microbiology aims to publish outstanding contributions to the understanding of interactions between microbes, prokaryotes and eukaryotes, and their host in the context of pathogenic or mutualistic relationships, including co-infections and microbiota. We welcome studies on single cells, animals and plants, and encourage the use of model hosts and organoid cultures. Submission on cell and molecular biological aspects of microbes, such as their intracellular organization or the establishment and maintenance of their architecture in relation to virulence and pathogenicity are also encouraged. Contributions must provide mechanistic insights supported by quantitative data obtained through imaging, cellular, biochemical, structural or genetic approaches.