基质金属蛋白酶-9和骨桥蛋白在甲状腺癌和腺瘤组织中的表达及其临床意义

目的 探讨基质金属蛋白酶-9 (MMP-9)和骨桥蛋白(OPN)在甲状腺癌及腺瘤组织中的表达和临床意义.方法 应用免疫组织化学法检测MMP-9和OPN在85例甲状腺癌组织、26例甲状腺瘤和15例正常甲状腺组织中的表达水平.结果 MMP-9在正常甲状腺、甲状腺瘤和甲状腺癌组织中的表达率分别为20.00％ (3/15)、53.85％ (14/26)和84.71％ (72/85),两两比较差异有统计学意义(P＜0.05),MMP-9表达水平与甲状腺癌美国肿瘤联合委员会(AJCC)分期、组织分化、颈部淋巴结转移、包膜浸润明显相关(P＜0.05);OPN在正常甲状腺、甲状腺瘤和甲状腺癌组织中的表达率分别为13.33％ (2/15)、46.15％ (12/26)和74.12％ (63/85),两两比较差异有统计学意义(P＜0.05),OPN表达水平与甲状腺癌AJCC分期、颈部淋巴结转移、包膜浸润明显相关(P＜0.05).结论 检测MMP-9和OPN有助于判断甲状腺癌浸润转移潜能.     

骨桥蛋白在哮喘小鼠肺组织中的表达及地塞米松的干预作用

目的 探讨哮喘气道炎症与骨桥蛋白(OPN)表达的相关性,以及地塞米松(DXM)对OPN表达的影响.方法 50只小鼠随机分为正常对照组、不同程度哮喘组(卵清蛋白OVA激发1周和2周组)及DXM治疗1周和2周组,每组10只;OVA致敏、激发建立急性哮喘模型;苏木精-伊红染色观察各组小鼠气道炎症情况;ELISA法检测血清OVA-sIgE水平;免疫组化和Western blot法分别定位和定量分析肺组织OPN表达;Real-time PCR检测肺组织OPN mRNA水平.结果 哮喘组气道病理学改变较对照组和DXM组明显,且OVA激发2周哮喘组改变较激发1周组明显.哮喘组血清OVA-sIgE水平较对照组和DXM组明显升高(PPPP结论 OPN可能是哮喘发病机制中的重要因素,哮喘气道炎症越重,OPN表达水平可能越高;DXM可能通过抑制OPN的表达来减轻哮喘炎症.     

釉基质蛋白体外诱导大鼠外胚间充质细胞骨桥素和骨钙素mRNA的表达

目的：了解体外培养环境中，大鼠外胚间充质细胞在两种不同形式釉基质蛋白(粗提型EMPs及纯化型EMD)诱导条件下，成骨分化标志骨桥素及骨钙素mRNA的表达水平。方法：酶消化法原代培养SD大鼠颌突外胚间充质细胞，在含釉基质蛋白(EMPs或EMD或EMPs EMD)的培养液中进行体外诱导培养，提取总RNA，采用RT-PCR法，以β-actin cDNA为内参照，检测骨桥素及骨钙素mRNA的表达水平。结果：1)各诱导实验组细胞均出现骨桥素mRNA的表达；2)仅EMPs EMD组出现骨钙素mRNA的表达。结论：外胚间充质细胞经釉基质蛋白特别是粗提型EMPs及纯化型EMD体外复合和诱导培养，可向成骨或成牙骨质方向分化，出现骨桥素及骨钙素mRNA的表达。     

Protective effect of ellagic acid on healing alveolar bone after tooth extraction in rat—A histological and immunohistochemical study

Objective

This study has attempted to evaluate the effects of ellagic acid (EA) on alveolar bone healing after tooth extraction in rats.

Design

Twenty-four Sprague Dawley (SD) male rats (200–250 g) were selected and were anaesthetised for the extraction of upper left incisor. Then, the rats were divided into two groups, comprising 12 rats each; the first group has been considered as a control group and was given only normal saline, whereas, the second group (treated group) was intragastrically administrated with EA daily once, for 28 days. Then three rats from each group had been selected on 7th, 14th, 21st, and 28th days to dissect their maxilla tissue either for histological observation and homogenisation purposes. The tissues fixed, decalcified and embedded in paraffin. Serial sections of 5 μm thickness were prepared and stained with haematoxylin and eosin (H&E) for the histological study. Similar sections were taken for immunohistochemical analysis to assess osteocalcin (OSC) and osteopontin (OPN). Furthermore, Malondialdehyde (MDA) and superoxide dismutase (SOD) were measured in homogenated gingival maxilla tissue of rat by commercial kit.

Results

Based on the histological analysis we have identified that, EA treatment has induced earlier trabecular bone deposition in the treated group, resulting in more organised bone matrix on the 14th, 21st, and 28th days after tooth extraction, as against the control group. In comparison to control group, the positive labelling of OSC and OPN of the treated group have been highly expressed in the alveolar socket on 14th, and 21st days, which has indicated a the possibility of formation of new bone trabeculae at the beginning of the mineralisation process, after tooth extraction. In the EA treatment group, lipid per-oxidation (MDA) was significantly decreased (P < 0.05), as opposed to the control group. However, the antioxidant defense enzyme (SOD) was significantly increased in the maxilla tissue treated with EA (P < 0.05), compared to control group, which suggests that, after tooth extraction, EA plays an important role in the protection against the induction of lipid per-oxidation, particularly after 28 days of treatment with EA.

Conclusion

This study has concluded that, EA may accelerated the healing process in teeth socket of rats. Furthermore, the EA treated group showed a stronger positive immunolabelling for OSC and OPN, when compared with the control group.     

环孢素 A 对大鼠牙槽骨内 DMP1 C 末端和 OPN 表达的影响

目的：观察环孢素A（cyclosporinA,CsA）对大鼠牙槽骨内牙本质基质蛋白1（dentinmatrixprotein,DMP1）C末端和骨桥蛋白（osteopontin,OPN）表达的影响。方法：30只7周龄健康雄性Wistar大鼠,随机等分为对照组和实验组,每组再随机等分为20d、30d和40d3个亚组,实验组大鼠喂饲CsA（30mg／k·d）。免疫组化法染色下颌第一磨牙颊舌向石蜡切片,光镜下观察DMP1C末端和OPN在牙槽骨内的表达情况。结果：与对照组大鼠比较,实验组大鼠牙槽骨内DMP1C末端表达明显下调,OPN表达明显上调;在本实验周期内,上述变化随实验时间的延长而加重。结论：CsA影响大鼠牙槽骨内矿化相关非胶原蛋白DMP1C末端和OPN的表达,推测CsA可能抑制大鼠牙槽骨的矿化。     

骨桥蛋白与动脉粥样硬化和骨质疏松的关系

骨桥蛋白作为体内广泛存在的细胞因子和趋化因子, 可介导细胞粘附,聚集及迁移,具有抗炎和促炎的双重作用,近年来逐渐认识到它在心血管疾病中起着不可估量的作用。骨桥蛋白可能刺激成骨细胞增殖、钙化,促进破骨细胞与骨基质的黏附并提高破骨细胞的溶骨活性,刺激骨吸收及骨重建,增加骨吸收陷窝的数量和面积,并抑制骨组织的矿化过程。本文就骨桥蛋白在骨质疏松症与动脉硬化的发病中所起作用作一综述。     

PI3K/PKB信号通路在TGF-β_1诱导人肝星状细胞表达骨桥蛋白中的作用

目的:研究磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/PKB)信号通路在转化生长因子β1(TGF-β1)诱导人肝星状细胞表达骨桥蛋白(OPN)的调控作用。方法:体外培养LX-2人肝星状细胞株,予TGF-β1(终浓度2.5、5、10、20μg/L)刺激24 h或予TGF-β1(终浓度10μg/L)刺激12 h、24 h、48 h;先经PI3K/PKB信号通路特异性抑制剂wortmannin(0.1μmol/L)预处理1 h,再予10μg/L TGF-β1刺激24 h,收集细胞,采用real-time PCR及Western blotting法检测OPN表达情况。结果:TGF-β1能够促进LX-2细胞表达OPN,在一定浓度和时间范围内,其表达量随着TGF-β1浓度和时间的增加而增加,呈剂量和时间依赖性关系;经wortmannin预处理再予TGF-β1刺激的LX-2细胞,与对照组相比,OPN表达受到明显抑制(P0.01)。结论:TGF-β1对LX-2人肝星状细胞OPN表达具有诱导作用,此作用可能受PI3K/PKB信号通路的调控。     

骨桥蛋白通过内质网应激诱导C2C12肌细胞胰岛素抵抗

目的:探讨骨桥蛋白(osteopontin,OPN)对C2C12肌细胞胰岛素抵抗的影响及其可能的机制。方法:低血清培养辅以胰岛素处理诱导C2C12成肌细胞分化为C2C12肌细胞,蛋白免疫印迹检测蛋白质的表达,离心法分离细胞膜,葡萄糖摄取试剂盒定量葡萄糖摄取。结果:(1)骨桥蛋白以剂量依赖和时间依赖方式抑制胰岛素刺激的蛋白激酶B(Akt)的磷酸化,并抑制胰岛素所致的葡萄糖转运体4(Glut4)膜位移和葡萄糖的摄取;而特异性的抗OPN受体CD44抗体预处理可逆转上述变化。(2)OPN可诱导C2C12肌细胞的内质网应激,并促进c-Jun氨基端激酶(JNK)的磷酸化。(3)内质网应激抑制剂4-苯基丁酸(4-PBA)可降低OPN所增加的JNK磷酸化,恢复胰岛素刺激所致的Glut4的膜位移以及葡萄糖摄取。结论:骨桥蛋白通过诱导C2C12肌细胞内质网应激导致胰岛素抵抗。本研究为揭示骨桥蛋白在胰岛素抵抗和糖尿病中的作用提供了新的实验依据和理论解释。     

gax基因转染对血管平滑肌细胞中基质金属蛋白酶2和骨桥蛋白转录的影响

目的:探讨腺病毒介导gax基因转染血管平滑肌细胞(VSMC)后,VSMC中基质金属蛋白酶2(MMP2)和骨桥蛋白基因转录的变化。方法:以携带大鼠gax基因表达序列的复制缺陷型5型腺病毒载体(AdCMV-gax)转染VSMC后,应用免疫细胞化学染色检NGax蛋白表达的变化,以RT-PCR技术检测VSMC中MMP2 mRNA和骨桥蛋白mRNA的变化。结果:AdCMV-gax转染后:(1)VSMC中Gax蛋白的表达比转染前显著增高(P<0.01); (2)VSMC中MMP2 mRNA和骨桥蛋白mRNA 比未转染组均显著降低(P<0.05)。结论:增强gax基因表达可抑制MMP2和骨桥蛋白的转录。