关键词： asporin   CXCR4   DDR2   gut-microbiome   leptin   osteoarthritis   SDF-1  

摘要： Osteoarthritis (OA), a disorder of joints, is prevalent in older age. The contemporary cure for OA is aimed to confer symptomatic relief, consisting of temporary pain and swelling relief. In this paper, we discuss various modalities responsible for the onset of OA and associated with its severity. Inhibition of chondrocytes receptors such as DDR2, SDF-1, Asporin, and CXCR4 by specific pharmacological inhibitors attenuates OA, a critical step for finding potential disease modifying drugs. We critically analyzed recent OA studies with an emphasis on intermediate target molecules for OA intervention. We also explored some novel and safe treatments for OA by considering disease prognosis crosstalk with cellular signaling pathways.

关键词： Lung cancer   Fibroblasts   SCRIB   Invasion   Asporin  

摘要： Aims: Cancer associated fibroblasts (CAFs) play a crucial role in lung tumor development, but the underlying mechanism is still not fully understood. Main methods: SCRIB expression in the CAFs of human lung cancer tissues was examined by immunohistochemistry (IHC). A coculture of mouse Lewis lung cancer cells (LLC) and fibroblasts was used to investigate SCRIB expression in cocultured fibroblasts. Proliferation, scratch wound, and transwell assays were used to examine the proliferation, migration and invasion ability of SCRIB knockdown fibroblasts and their effects on LLC. A 3D-coculture system and co-injection xenograft model were used to examine LLC invasion. RNA sequencing and transwell experiments were used to explore the molecules that may participate in LLC invasion. Key findings: Herein, we found that the low expression of SCRIB in CAFs is correlated with advanced tumor stages and poor survival for human lung squamous cell carcinoma. SCRIB expression in fibroblasts is drastically downregulated by LLC cells. SCRIB knockdown fibroblasts not only enhance invasion but also facilitate LLC invasion in a 3D-coculture system and in an in vivo subcutaneous transplantation model. The upregulation of asporin in SCRIB knockdown fibroblasts is involved in LLC invasion in vitro. Significance: Collectively, the results indicate that fibroblasts with low SCRIB expression promote lung cancer cell invasion, which suggests that the downregulated expression of SCRIB may represent one of the important characteristics of tumor-promoting CAFs in lung squamous cell cancer.

关键词： asporin   cardiomyocyte apoptosis   glycated low-density lipoproteins   grape seed procyanidin B2   transforming growth factor-beta 1  

摘要： The progression of diabetic cardiomyopathy is related to cardiomyocyte dysfunction and apoptosis. Our previous studies showed that asporin (ASPN) was significantly increased in the myocardium of db/db mice through proteomics, and grape seed procyanidin B2 (GSPB2) significantly inhibited the expression of ASPN in the heart of db/ db mice. We report here that ASPN played a critical role in glycated low-density lipoproteins (gly-LDL) induced-cardiomyocyte apoptosis. We found that gly-LDL upregulated ASPN expression. ASPN increased H9C2 cardiomyocyte apoptosis with down-regulation of Bcl-2, upregulation of transforming growth factor-beta 1, Bax, collagen III, fibronectin, and phosphorylation of smad2 and smad3. However, GSPB2 treatment reversed ASPN-induced impairments in H9C2 cardiomyocytes. These results provide evidence for the cardioprotective action of GSPB2 against ASPN injury, and thus suggest a new target for fighting against diabetic cardiomyopathy.

关键词： Periodontal   Mesenchymal stem cells   Asporin   Periostin   Cementum Protein-1   Bone marrow  

摘要： Background: Periodontal ligament stem cells are a source of mesenchymal stem cells, but it is unclear whether their phenotype is distinct from mesenchymal stem cells derived from different tissues, such as those derived from bone marrow. Objective: To investigate the expression of the putative PDL markers asporin, periostin, nestin and cementum protein 1, by periodontal ligament stem cells both constitutively and during osteogenic differentiation when compared to bone marrow-derived mesenchymal stem cells, and dental pulp stem cells. Methods: The primary human periodontal ligament, bone marrow, and dental pulp stem cells, and osteoblasts from different donors were cultured in vitro. The expression of periodontal marker associated genes during osteogenic induction was tested by qRT-PCR and immunofluorescence staining. Results: Asporin expression was detected in periodontal ligament stem cells and increased markedly during the time in culture (upregulated x53 fold at 21 days post-induction). During osteogenic differentiation, asporin expression significantly decreased in periodontal ligament cells whereas periostin significantly decreased in dental pulp cells. Periostin expression was absent in osteoblasts, but expression gradually increased in all other cells with time in culture. Nestin expression was mainly seen in the periodontal ligament and dental pulp cells and was largely absent in osteoblasts and bone marrow cells. Cementum protein-1 was most highly expressed in bone marrow cells and osteoblasts following osteogenic induction. Conclusions: The results provide further evidence that periodontal ligament-derived and bone marrow derived mesenchymal stem cells are phenotypically distinct. Periodontal markers are also expressed in dental pulp stem cells.

摘要： Worldwide, calcific aortic valve disease is one of the leading causes of morbidity and mortality among patients with cardiac abnormalities. Aortic valve mineralization and calcification are the key events of adult calcific aortic valve disease manifestation and functional insufficiency. Due to heavy mineralization and calcification, adult aortic valvular cusps show disorganized and dispersed stratification concomitant with deposition of calcific nodules with severely compromised adult valve function. Interestingly, shared gene regulatory pathways are identified between bone-forming cells and heart valve cells during development. Asporin, a small leucine-rich proteoglycan (43 kDa), acts to inhibit mineralization in periodontal ligament cells and is also detected in normal murine adult aortic valve leaflets with unknown function. Therefore, to understand the Asporin function in aortic cusp mineralization and calcification, adult avian aortic valvular interstitial cell culture system is established and osteogenesis has been induced in these cells successfully. Upon induction of osteogenesis, reduced expression of Asporin mRNA and increased expression of bone and osteogenesis markers are detected compared to cells maintained without osteogenic induction. Importantly, treatment with human recombinant Asporin protein reduces the mineralization level in osteogenic media-induced aortic valvular interstitial cells with the concomitant decreased level of Wnt/β-catenin signaling. Overall, all these data are highly indicative that Asporin might be a novel biomolecular target to treat patients of calcific aortic valve disease over current cusp replacement surgery. © 2020 Anisha Polley et al.

关键词： 乳头状甲状腺癌   TMT定量蛋白质组学   Asporin   HER2   肿瘤侵袭和转移  

摘要： 甲状腺癌是内分泌系统最常见的恶性肿瘤。过去二十年,甲状腺癌新发病例在全球呈现“潮涌”态势,已成为各个国家及地区发病增长速度最快的恶性肿瘤之一。84%的甲状腺癌是乳头状甲状腺癌(Papillary thyroid cancer,PTC),对小于45岁PTC病人,淋巴结转移是预后变差的独立危险因素尚存在争议;但对于大于45岁PTC病人,学术界普遍认为这类病人的淋巴结转移与其高死亡率、手术后复发及预后差正相关。特别是与无淋巴结转移(N0)或Ⅵ区淋巴结转移(N1a)相比,发生Ⅰ-Ⅵ区淋巴结转移(N1b)的PTC病人具有更高的术后复发率。遗憾的是,目前尚未有研究报道大于45岁PTC病人淋巴结转移的分子机制。为此,本课题从两方面展开研究:第一,分析PTC-N0、PTC-N1a和PTC-N1b癌组织之间蛋白水平和分子功能的差异;第二,分析PTC-N0、PTC-N1a和PTC-N1b癌与癌旁正常组织之间蛋白水平和分子功能的差异,并深入研究潜在的调控机制,寻找新的肿瘤标志物。第一部分研究结果:通过非监督的主成分和火山图分析发现,大于45岁的PTC病人,淋巴结转移程度不同,癌组织中蛋白的表达也明显不同,而癌旁正常组织中蛋白的表达差异却很小。通过热图、信号通路和基因本体富集分析PTC-N0、PTC-N1a和PTC-N1b 癌组织中的差异表达蛋白,发现这些差异表达蛋白主要富集于细胞外基质、能量代谢和细胞增殖等生物学过程,从宏观蛋白层面揭示PTC-N0、PTC-N1a和PTC-N1b预后不同的可能分子机制。第二部分研究结果:与癌旁正常组织相比,PTC-N0、PTC-N1a和PTC-N1b癌组织中蛋白的表达模式不甚相同。通过生物信息学工具筛选出PTC-N0、PTC-N1a和PTC-N1b癌组织中共同高表达的差异蛋白,并从中挑选出5个进行Western Blotting 验证,发现 Versican、PL3、SERP1NA1、CD55 和 Asporin 蛋白在 PTC-N0、PTC-N1a和PTC-N1b癌组织中均高表达。免疫组化实验发现,相比癌旁正常组织,Asporin在PTC癌组织中明显高表达,且主要表达于细胞外基质、细胞膜及胞浆。TCGA THCA数据库验证发现,PTC癌组织中高表达的Asporin mRNA与大直径肿瘤,淋巴结转移,双叶占位,高TNM分期和BRAFV600E突变,总生存期和无进展生存期缩短正相关。以上结果提示Asporin在PTC中可能是个促癌基因,可促进PTC增殖、侵袭和转移。CCK8实验发现,敲低Asporin后,BCPAP和KTC-1细胞活力明显降低。克隆形成实验发现,敲低Asporin后,BCPAP和KTC-1细胞克隆形成能力明显减弱,提示Asporin具有促进PTC癌细胞增殖和克隆形成的作用。Transwell实验发现,敲低Asporin后,BCPAP、KTC-1和BHT101细胞侵袭和转移能力都明显降低,提示Asporin具有促进甲状腺癌细胞侵袭和转移的作用。为了进一步研究Asporin致病的分子机制,Western Blotting发现,敲低BCPAP、KTC-1和BHT101细胞中Asporin后,MAPK信号通路中BRAF和p-ERK1/2蛋白表达降低,而t-ERK1/2蛋白表达不变,且MAPK信号通路下游的正向调控EMT相关蛋白β-catenin、SNAIL、SLUG、ZEB1 和 ZEB2 表达降低。提示 Asporin 可通过 MAPK 信号通路上调其下游EMT相关蛋白的表达水平。此外,敲低Asporin后,BCPAP和KTC-1细胞形态主要呈圆形,伪足减少,胞内微丝分布紊乱,提示Asporin可通过促进MAPK信号通路的激活,上调EMT相关蛋白的表达水平,进而引起细胞骨架的改变,最后促进PTC的侵袭和转移。免疫共沉淀实验发现,Asporin可结合MAPK信号通路上游蛋白HER2,而不结合EGFR和SRC。免疫荧光实验发现,Asporin和HER2在BCPAP和KTC-1细胞膜及胞浆中共定位,进一步提示Asproin和HER2存在相互作用。通过Western Blotting实验发现,相比对照细胞,Asporin敲低后,BCPAP和KTC-1细胞中HER2、p-HER2Y1248、p-SRCY418、p-EGFRY845 和 p-EGFRY1173 表达水平降低,而 SRC 和EGFR表达水平不变,提示Asporin可通过结合HER2,上调HER2表达水平,进而使p-EGFR和p-SRC表达升高,从而激活MAPK信号通路。体外rhAsporin刺激和PLX4032抑制实验发现,rhAsporin并不能在细胞外通过结合HER2激活MAPK信号通路,且MAPK信号通路并不能够正反馈调控Asporin蛋白表达水平

关键词： 椎间盘退变   腰背痛   再生治疗   生长因子   细胞治疗   基因治疗   病毒载体   非病毒载体   组织工程  

摘要： 背景:多因素引起的椎间盘退行性病变所导致的严重的腰背痛是致残率最高的脊柱疾病,随着医学分子生物学的发展及椎间盘退变机制研究的深入,椎间盘退变的再生疗法为恢复这种难治性疾病提够了可能。目的:文章对近年来国内外各研究者关于椎间盘退变的相关分子和各种再生疗法在椎间盘退变中应用的研究进行归纳总结。方法:使用计算机检索PubMed、Embase、Web of science 数据库和中国知网、万方、维普数据库在1995年1 月至2019 年3 月发表的关于椎间盘退变再生治疗的相关文献,其中中文的检索关键词包括“椎间盘退变、腰背痛、再生治疗、生长因子、细胞治疗、基因治疗、病毒载体、非病毒载体”,英文的关键词为“Intervertebraldisc degeneration,low back pain,regenerative therapy,growth factor,cell therapy;gene therapy,viralvector,non-viral vector”,筛选出相关文献后再进一步归纳总结。结果与结论:研究显示,近20 多种基因影响着髓核细胞的变性,其中就包括各种非编码RNA(如microRNA、lincRNA 和lncRNA)。目前,生长因子疗法、细胞疗法和基因疗法等再生研究已经取得了预期效果。非病毒载体转导的基因治疗能避免感染的风险和靶细胞突变的可能,将成为恢复或缓解椎间盘退变的强大工具,具用广阔的应用前景。然而,这些再生疗法也有一定的局限性,在其应用于临床之前,需进行大量的基础研究和临床实验,以便评估其疗效和对逆转退变的影响。

关键词： 椎间盘退变   生长因子治疗   细胞治疗   基因治疗   组织工程治疗  

摘要： 下腰痛是骨科常见的疾病之一,而椎间盘髓核退变是引起下腰痛的主要原因。目前对于椎间盘退变的治疗方式主要是保守治疗和手术治疗,然而这些治疗方法的目的是缓解症状,并非是针对椎间盘退变发生机制的病因治疗。对椎间盘髓核退变发生机制的详细了解有助于开展生物学再生修复。目前,针对髓核退变的分子机制和背景,发展出不同的髓核退变生物学再生修复的方法。文章从生长因子治疗、基因治疗、细胞治疗、组织工程化髓核等方面对髓核退变生物学再生修复进行综述。

关键词： asporin   collagen fiber   osteoclastogenesis   periodontitis   PLAP-1  

摘要： Background and ObjectivePeriodontalligament-associated protein-1 (PLAP-1) is an important regulator of osteogenic differentiation ofperiodontalligament cells and plays important role in the homeostasis of periodontal tissues. But the role of PLAP-1 in periodontitis is poorly understood. Expressions of PLAP-1 in experimental periodontitis are observed to elucidate whether PLAP-1 gets involved in the pathogenesis of periodontitis. Material and MethodsWistar rats were randomly allocated to two groups (n=6/group): Ligation group and Control group. PLAP-1 expression in experimental periodontitis was assessed by immunohistochemistry and collagen fibers in periodontal ligament were observed using picrosirius red staining. Expressions of PLAP-1 and CD68 in periodontitis were colocalized by double-labelled immunofluorescence. To further examine the relationship between PLAP-1 and osteoclastogenesis in experimental periodontitis, acute periodontal inflammatory infiltration and alveolar bone destruction were induced by administering ligated rats with 10ng/mL tumor necrosis factor alpha (TNF-;ligation+TNF- group, n=6). Alveolar bone loss was observed by micro-computed tomography (Micro-CT), and osteoclasts were identified by tartrate-resistant acid phosphatase staining (TRAP). Expressions of PLAP-1 in TNF- stimulated human periodontal ligament cells were also detected at 24 and 48hours by western blotting. ResultsPLAP-1 expression levels in periodontal ligament cells and collagen fibers were lower in the ligation group,compared with the control group. Similarly, TNF- decreased PLAP-1 expression in human periodontal ligament cells in vitro. Degradation or destruction of collagen fibers accompanied the reduced PLAP-1 expression in the periodontal ligament in the ligation group. Colocalization of PLAP-1 and CD68 revealedthe positive relationship between PLAP-1 and CD68+ infiltrating cells in periodontitis. More PLAP-1-positive inflammatory cells were found in the ligation+TNF- gro