Zhang Zhiguo, Chen Yanjing, Xiang Lihua, Wang Zhen, Xiao Gary Guishan, Ju Dahong
Institute of Basic Theory, China Academy of Chinese Medical Sciences, Beijing 100700, P.R. China.
School of Pharmaceutical Science, Dalian University of Technology, Dalian, Liaoning 116024, P.R. China.
Exp Ther Med. 2018 Nov;16(5):3939-3950. doi: 10.3892/etm.2018.6681. Epub 2018 Sep 3.
The present study assessed the changes in long non-coding (lnc)RNA and mRNA expression profiles when diosgenin (DIO) exerted a potential osteoprotective effect on the alveolar bone of ovariectomized (OVX) rats. Female Wistar rats underwent a sham operation (SHAM group) or ovariectomy. OVX rats were treated using vehicle (OVX group), DIO (DIO group) or estradiol valerate (EV group) for 12 weeks. After treatment, the biomarkers of bone turnover in plasma and the microstructure of alveolar bone were assessed. lncRNA microarrays were applied to assess lncRNA and mRNA expression profiles in alveolar bone in the OVX and DIO group rats. Subsequently, the differentially expressed mRNAs associated with the comprehensive bone metabolism pathway in Ingenuity Pathway Analysis (IPA) were identified and regarded as key mRNAs. Based on some of the key mRNAs and all the differentially expressed lncRNAs, a coexpression network was established and this network was further analyzed to identify the top 6 lncRNAs with the highest closeness scores (pivotal lncRNAs). Finally, 6 modules showing interactions between pivotal lncRNAs and key mRNAs were constructed. All of the pivotal lncRNAs and key mRNAs were validated with reverse transcription-quantitative polymerase chain reaction. The present findings demonstrated that DIO suppressed the loss of alveolar bone in OVX rats, and the changes to the expression of some lncRNAs or mRNAs occurred in the alveolar bone of the rats in the DIO group. Twenty-four key mRNAs were identified during pathway analysis. Furthermore, 8/24 key mRNAs ( and ) were used to establish a coexpression network, which included 1,656 nodes and 5,341 edges. During network analysis, 6 pivotal lncRNAs (XR_008346, MRuc007iji, MRAK157089, MRAK076413, MRAK143591 and AB036696) were obtained, and 6 modules illustrating pivotal lncRNA-key mRNA interactions were identified. These results revealed that the anti-osteoporotic effect of DIO on alveolar bone may be associated with the promotion of a bone formation process through increasing the signaling of the Wnt and BMPs pathways and the inhibition of the bone resorption process through decreasing stimulators of osteoclastogenesis. To conclude, several pivotal lncRNAs may serve important roles in these processes via regulating some key mRNAs in the bone metabolism pathway.
本研究评估了薯蓣皂苷元（DIO）对去卵巢（OVX）大鼠牙槽骨发挥潜在骨保护作用时，长链非编码（lnc）RNA和mRNA表达谱的变化。雌性Wistar大鼠接受假手术（假手术组）或卵巢切除术。OVX大鼠分别用赋形剂（OVX组）、DIO（DIO组）或戊酸雌二醇（EV组）治疗12周。治疗后，评估血浆中骨转换的生物标志物以及牙槽骨的微观结构。应用lncRNA微阵列评估OVX组和DIO组大鼠牙槽骨中的lncRNA和mRNA表达谱。随后，在 Ingenuity Pathway Analysis（IPA）中鉴定与综合骨代谢途径相关的差异表达mRNA，并将其视为关键mRNA。基于一些关键mRNA和所有差异表达的lncRNA，建立共表达网络，并对该网络进行进一步分析以鉴定紧密性得分最高的前6个lncRNA（关键lncRNA）。最后，构建了6个显示关键lncRNA与关键mRNA之间相互作用的模块。所有关键lncRNA和关键mRNA均通过逆转录定量聚合酶链反应进行验证。本研究结果表明，DIO抑制了OVX大鼠牙槽骨的丢失，并且DIO组大鼠牙槽骨中一些lncRNA或mRNA的表达发生了变化。在通路分析过程中鉴定出24个关键mRNA。此外，使用8/24个关键mRNA（和）建立共表达网络，该网络包含1656个节点和5341条边。在网络分析过程中，获得了6个关键lncRNA（XR_008346、MRuc007iji、MRAK157089、MRAK076413、MRAK143591和AB036696），并鉴定出6个说明关键lncRNA与关键mRNA相互作用的模块。这些结果表明，DIO对牙槽骨的抗骨质疏松作用可能与通过增加Wnt和BMPs途径的信号传导促进骨形成过程以及通过减少破骨细胞生成刺激因子抑制骨吸收过程有关。总之，一些关键lncRNA可能通过调节骨代谢途径中的一些关键mRNA在这些过程中发挥重要作用。
