绝经后骨质疏松患者血清I型胶原氨基末端、I型胶原羧基末端及骨钙素的表达变化及临床意义

摘要 目的：探讨绝经后骨质疏松患者血清I型胶原氨基末端（NTx）、I型胶原羧基末端（CTX）及骨钙素（BGP）的表达变化及临床意义。方法：选取我院2017年8月-2022年8月收治的60例绝经后骨质疏松患者作为研究对象,将其分为观察组,另选取同期来我院体检的60名绝经后健康志愿者作为对照组。对比两组患者NTx、CTX、BGP表达水平,并建立受试者特征工作（ROC）曲线分析NTx、CTX、BGP对绝经后骨质疏松的诊断效能。3个月后对所有患者进行门诊复查随访,将症状明显明显减轻,X线检查明显改善,骨密度值明显增加的35例绝经后骨质疏松患者分为预后良好组,将其余25例未达到上述标准的患者分为预后不良组,对比两组患者临床一般情况,并应用Logistic回归分析NTx、CTX、BGP对绝经后骨质疏松的预后预测价值。结果：两组受检者NTx、CTX、BGP表达水平对比差异显著,观察组NTx、CTX高于对照组,BGP低于对照组（P＜0.05）; NTx、CTX、BGP三者联合对绝经后骨质疏松的诊断效能优于单一检测（P＜0.05）;预后良好组与预后不良组患者年龄、BMI、合并基础疾病、病程、Ca表达水平对比无明显差异（P＞0.05）,预后良好组与预后不良组患者病情严重程度、骨密度T值、雌二醇、血清NTx、CTX、BGP表达水平对比差异显著（P＜0.05）;logistic回归分析结果表明：CTX、BGP为绝经后骨质疏松的预后不良的独立影响因素（P＜0.05）。结论：绝经后骨质疏松患者血清I型胶原氨基末端、I型胶原羧基末端表达水平高于非骨质疏松群体,骨钙素低于非骨质疏松群体,三者联合可提升绝经后骨质疏松的诊断效能。另外,CTX、BGP作为绝经后骨质疏松的预后不良的独立影响因素,CTX水平越高、BGP水平越低可能预示患者预后不良,因此临床需针对此类患者及时改良治疗措施,提升其预后水平。     

Insulin-like growth factor-I attenuates the inhibitory effect of type I collagen through beta1 integrin receptor

Extracellular matrix and growth factors are the crucial factors that regulate healing and regenerating processes in human periodontal ligament cells. The purpose of this study was to examine the effects of type I collagen and insulin-like growth factor-I (IGF-I) on osteopontin (OPN) expression. The data showed that OPN expression was significantly decreased when cells were cultured on collagen-coated plates. Addition of IGF-I obviously induced OPN expression only in a collagen-coated condition, suggesting an attenuating effect of IGF-I on the decrease of OPN expression. Cells treated with a combination of inhibitory antibody to beta1 integrin and IGF-I showed the same level of OPN expression as those treated with either inhibitory antibody to beta1 integrin or IGF-I alone. These results indicate that IGF-I counteracts with the inhibitory signal from type I collagen through beta1 integrin receptor.     

Expression of insulin-like growth factor-1 and insulin-like growth factor-1 receptors in EL4 lymphoma cells overexpressing growth hormone

Almost all of the previous studies with growth hormone (GH) have been done with exogenously supplied GH and, therefore, involve actions of the hormone through its receptor. However, the actions of endogenous or lymphocyte GH are still unclear. In a previous study, we showed that overexpression of GH (GHo) in a lymphoid cell line resulted in protection of the cells to apoptosis mediated by nitric oxide (NO). In the present study, we show that the protection from apoptosis could be transferred to control cells with culture fluids obtained from GHo cells and blocked by antibodies to the insulin-like growth factor-1 (IGF-1) or antibodies to the IGF-1-receptor (IGF-1R). Northern and Western blot analysis detected significantly higher levels of IGF-1 in cells overexpressing GH. An increase in the expression of the IGF-1R in GHo cells was also detected by Western blot analysis, (125)I-IGF-1 binding and analysis of IGF-1R promoter luciferase constructs. Transfection of GHo cells with a dominant negative IGF-1R mutant construct blocked the generation of NO and activation of Akt seen in GHo cells compared to vector alone control EL4 cells. The results suggest that one of the consequences of the overexpression of GH, in cells lacking the GH receptor, is an increase in the expression of IGF-1 and the IGF-1R which mediate the protection of EL4 lymphoma cells from apoptosis.     

子宫内膜癌中TGF-β1及受体TβRI的表达及其临床意义

目的：研究转化生长因子（TGF-β1）及其Ⅰ型受体（TβRI）在子宫内膜癌组织中的表达及其临床意义。方法：采用RT-PCR方法检测36例子宫内膜癌及31例正常子宫内膜组织中TGF-β1及TβRI基因的表达水平,分析其与临床病理参数之间的关系。结果：子宫内膜癌组织中TGF-β1mRNA的表达明显高于正常组织（P〈0.05）,而TβRI mRNA的表达水平明显低于正常内膜组织（P〈0.05）。TGF-β1mRNA及TβRImRNA在子宫内膜癌中的表达均与肿瘤的临床分期、组织学分级、肌层浸润深度、淋巴结转移呈显著性相关（P〈0.05）,差别有统计学意义。结论：TGF-β1及其Ⅰ型受体TβRI在子宫内膜癌的发生、发展中可能起重要作用,检测TGF-β1及TβRI对评价子宫内膜癌的转移及预后有重要价值。     

Influence of exercise duration on serum insulin-like growth factor and its binding proteins in athletes

The changes in circulating concentrations of insulin-like growth factors during exercise have to date remained incomplete in their documentation. Therefore, we examined in 25 healthy athletes the effects of three different durations of three types of exercise – incremental ergometer cycling exercise (ICE), long-distance Nordic ski race (NSR) and a treadmill-simulated soccer game (TSG) lasting 20 min, 3 h, and 2 × 45 min separated by a 15-min half-time rest respectively, on plasma concentrations of growth hormone ([GH]), insulin-like growth factor-1 ([IGF-I]) and its binding proteins 1 and 3 ([IGFBP-1], [IGFBP-3]). Compared to baseline, serum [GH] increased by 15.2-fold after ICE (P < 0.001), 2.9-fold after NSR (P < 0.01) and 4.6-fold after TSG. Serum [IGF-I] rose by 11.9% after ICE (P < 0.001), while it decreased by −14.6% after NSR (P < 0.001) and was unchanged after TSG. Serum [IGFBP-1] was slightly increased (1.7-fold) after ICE (P < 0.01), but increased markedly (11.8-fold) after NSR (P < 0.001) and by 6.3-fold after the second session of TSG (P < 0.01) (it remained unchanged at the end of the first period of TSG, i.e. after 45-min exercise). The [IGFBP-3] increased by 14.7% after ICE (P < 0.001) and by 6% after TSG (P < 0.05) while it did not change after NSR. From our results it would appear that [IGFBP-1] increase to bind free IGF and hinder their insulin-like action during long-term exercise (lasting beyond 45 min). It is suggested that IGFBP-1 might thus contribute both to preventing hypoglycaemic action of IGF and to facilitating glucose uptake by muscle cells when muscle glycogen stores become deplete. Accepted: 27 May 1998     

TGF-beta1通过抑制HIF-1alpha减少风湿性心脏病心肌细胞胶原合成

目的:探讨缺氧诱导因子-1α(HIF-1α)在转化生长因子β1(TGF-β1)促风湿性心脏病心肌细胞胶原合成中的作用。方法:以体外培养风湿性心脏病(风心病)患者经瓣膜置换术后留取组织分离而来的心肌细胞为研究对象,依据加入TGF-β1的浓度将前期实验分为四组:0,5,10及20(μg/L),观察TGF-β1对风湿性心脏病心肌细胞胶原合成及HIF-1α表达的影响;而后实验选取10μg/L TGF-β1为干预浓度,将Scrambled si RNA或HIF-1αsi RNA转染入细胞内。48小时后,分别收集各组细胞,采用RT-PCR检测I型胶原的m RNA水平,Western Blot技术测定细胞内I型胶原和HIF-1α的蛋白表达水平。结果:与0、5及10μg/L浓度TGF-β1组相比,5、10及20μg/L浓度的TGF-β1分别显著地增加了风心病心肌细胞I型胶原及HIF-1α的表达。另外,HIF-1αsi RNA则明显减少了TGF-β1诱导的心肌细胞I型胶原生成。结论:HIF-1α介导了TGF-β1对风湿性心脏病心肌细胞I型胶原合成的促进作用。     

Overexpressed homeobox B9 regulates oncogenic activities by transforming growth factor-β1 in gliomas

Glioma is the leading cause of deaths related to tumors in the central nervous system. The mechanisms of gliomagenesis remain elusive to date. Homeobox B9 (HOXB9) has a crucial function in the regulation of gene expression and cell survival, but its functions in glioma formation and development have yet to be elucidated. This study showed that HOXB9 expression in glioma tissues was significantly higher than that in nontumor tissues. Higher HOXB9 expression was also significantly associated with advanced clinical stage in glioma patients. HOXB9 overexpression stimulated the proliferation, migration, and sphere formation of glioma cells, whereas HOXB9 knockdown elicited an opposite effect. HOXB9 overexpression also increased the tumorigenicity of glioma cells in vivo. Moreover, the activation of transforming growth factor-β1 contributed to HOXB9-induced oncogenic activities. HOXB9 could be used as a predictable biomarker to be detected in different pathological and histological subtypes in glioma for diagnosis or prognosis.     

Angiotensin II type 1 receptor blockade attenuates renal fibrogenesis in an immune-mediated nephritic kidney through counter-activation of angiotensin II type 2 receptor

The relative roles of angiotensin II (Ang II) type 1 receptor (AT(1)R) and Ang II type 2 receptor (AT(2)R) in immune-mediated nephritis are unknown, and the effect of the blockade of AT(1)R and its indirect counter-activation of AT(2)R relative to the anti-fibrotic action in this disease is unclear. To address this question, we studied the role of AT(1)R and AT(2)R in anti-glomerular basement membrane nephritis in SJL mice. Groups of mice were treated with either an AT(1)R antagonist (CGP-48933; CGP group), an AT(2)R antagonist (PD-123319; PD group), both (CGP/PD group), or a vehicle (PCt group) from Day 29 to 56. At Day 56 post-treatment, fibrosis-related parameters such as interstitial matrix deposition, and the expression of genes of TGF-beta1, plasminogen activator inhibitor-1, and type I collagen were significantly reduced in the kidney in the CGP group. There were no significant effects on these parameters in the PD group. However, this anti-fibrotic action by CGP-48933 was totally abolished by co-treatment with PD-123319 in the CGP/PD group. The gene expression of renin was significantly increased in the kidneys in the CGP and CGP/PD groups, suggesting that CGP-48933 had increased Ang II generation in those groups. In conclusion, counter-activation of AT(2)R by increased Ang II under AT(1)R blockade likely conferred an anti-fibrotic protection in this model.     

血清cTnI、CK-MB及IGF-1在新生儿高胆红素血症中的变化及临床意义

摘要 目的：分析血清肌钙蛋白T( troponin T,cTnT)、肌酸激酶同工酶(creatine kinase isoenzyme,CK-MB)及胰岛素样生长因子-1(insulin-like growth factor-1,IGF-1)在新生儿高胆红素血症中变化及临床价值。方法：选择2015年1月至2019年12月我院新生儿科收治的80例新生儿高胆红素血症足月新生儿(观察组)以及80例非胆红素足月新生儿(对照组)为研究对象。收集住院资料,包括所有患儿的胎龄、日龄、出生体重、血清总胆红素（total bilirubin,TBIL）、CK-MB、cTnT、IGF-1、头颅核磁共振(skull magnetic resonance,MRI)、临床表现等及出院后随访资料。根据胆红素水平高低,将观察组分为轻度(33例)、中度组(27例)及重度组(20例),比较其：TBIL、IGF-1、CK-MB、cTnT水平。结果：各组新生儿TBIL、CK-MB、cTnT水平比较差异具有统计学意义(P＜0.05)。重度黄疸组TBIL、CK-MB、cTnT值分别高于轻度、中度黄疸组和对照组,IGF-1低于轻度、中度黄疸组和对照组(P＜0.05);中度黄疸组TBIL、CK-MB、cTnT高于轻度黄疸组和对照组,IGF-1低于轻度黄疸组和对照组(P＜0.05);轻度黄疸组TBIL、CK-MB、cTnT高于对照组,IGF-1低于对照组(P＜0.05)。血清cTnT与TBIL水平呈正相关(r=0.587,P＜0.05);血清CK-MB与TBIL水平无明显相关性(r=0.220,P＞0.05);血清IGF-1与TBIL水平呈负相关（r=-0.568,P＜0.05）。苍白球异常信号组患儿血清IGF-1值为(11.05±0.51) ng/mL,明显低于苍白球正常信号组(14.22±2.67) ng/mL(P＜0.05);苍白球异常信号组患儿血清TBIL水平为(347.62±33.01)μmol/L与苍白球正常信号组(341.75±35.14)μmol/L无明显差别(P＞0.05)。结论：检测血清cTnT和IGF-1水平分别有助于预测新生儿高胆红素血症心肌损伤和脑病的发生。     

Alpha-lipoic acid inhibits hepatic PAI-1 expression and fibrosis by inhibiting the TGF-β signaling pathway

Accumulating evidence suggests that plasminogen activator inhibitor (PAI)-1 plays an important role in the development of hepatic fibrosis via its involvement in extracellular matrix remodeling. We previously reported that alpha-lipoic acid (ALA), a naturally occurring thiol antioxidant, prevents hepatic steatosis by inhibiting the expression of sterol regulatory element binding protein-1c. The aim of the present study was to determine whether ALA prevents hepatic PAI-1 expression and fibrosis through the inhibition of multiple TGF-β-mediated molecular mediators. We investigated whether ALA inhibited the development of hepatic fibrosis in mice following bile duct ligation (BDL), an established animal model of liver fibrosis. We found that ALA markedly inhibited BDL-induced hepatic fibrosis and PAI-1 expression. We also found that ALA attenuated TGF-β-stimulated PAI-1 mRNA expression, and inhibited PAI-1 promoter activity in liver cells; this effect was mediated by Smads and the JNK and ERK pathways. The results of the present study indicate that ALA inhibits hepatic PAI-1 expression through inhibition of TGF-β-mediated molecular mediators, including Smad3, AP1, and Sp1, and prevents the development of BDL-induced hepatic fibrosis. These findings suggest that ALA may have a clinical application in preventing the development and progression of hepatic fibrosis.     

Recombinant human elafin promotes alveologenesis in newborn mice exposed to chronic hyperoxia

Background/aimsElastase inhibitors reverse elastin degradation and abnormal alveologenesis and attenuate the lung structural abnormalities induced by mechanical ventilation with O₂-rich gas. The potential of these molecules to improve endothelial function and to ameliorate severe bronchopulmonary dysplasia (BPD) during lung development is not yet understood. We sought to determine whether the intratracheal treatment of newborn mice with the elastase inhibitor elafin would prevent hyperoxia-induced lung elastin degradation and the cascade of events that cause abnormal alveologenesis.MethodsNewborn mice were exposed to 85% O₂ for 3, 7, 14 or 21 days. Recombinant human elafin was administered by intratracheal instillation from the first day every two days for 20 days. We next used morphometric analyses, quantitative RT-PCR, immunostaining, Western blotting, and ELISA methods to assess the key variables involved in elastogenesis disruption and the potential signaling pathways noted below in recombinant human elafin-treated mouse pups that had been exposed to 85% O₂.ResultsWe found that impaired alveolar development and aberrant elastin production were associated with elevations in whole lung elastase levels in 85% O₂-exposed lungs. Elafin attenuated the structural disintegration that developed in the hyperoxia-damaged lungs. Furthermore, elafin prevented the elastin degradation, neutrophil influx, activation of TGF-β1 and apoptosis caused by 85% O₂ exposure.ConclusionsPulmonary elastase plays an important role in disrupting elastogenesis during O₂-induced damage, which is the result of a pulmonary inflammatory response. Elafin prevents these changes by inhibiting elastase and the TGF-β1 signalling cascade and may be a new therapeutic target for preventing O₂-induced lung injury in neonates.     

TGF-beta1 induces alveolar epithelial to mesenchymal transition in vitro

The aim of this study was to investigate whether transforming growth factor-beta1 (TGF-beta1) could induce alveolar epithelial to mesenchymal transition (EMT) in vitro. Alveolar epithelial cells (AECs) from SD rats were isolated by elastase cell dispersion and IgG panning. Expression of alpha-smooth muscle actin (alpha-SMA) was assayed using Western blotting and immunostaining analysis. Morphological changes, the markers of epithelial cell (E-cadherin), and stress fiber by actin reorganization were detected by an indirect immunostaining. The contents of collagen I were determined by spectrophotometry. The levels of endogenous TGF-beta1 were measured with ELISA. Incubation of AECs with TGF-beta1 (0.1 approximately 10 ng/mL) induced abundant expression of alpha-SMA protein, and alpha-SMA expression in AECs reached a plateau when TGF-beta1 was > 3 ng/mL. Furthermore, we found that TGF-beta1 (3 ng/mL) exposure of AECs induced an authentic EMT characterized by abundant expression of alpha-smooth muscle actin, transformation of myofibroblastic morphology, increased formation of stress fiber by actin reorganization, and loss of epithelial marker E-cadherin. Meanwhile, significant increase in the levels of collagen I from 32.0 +/- 6.6 mg/g in control to 98 +/- 10.8 mg/g in TGF-beta1-treated group was found over a 72 h incubation period. Moreover, following stimulated by TGF-beta1 (3 ng/mL), a marked and time-dependent increase in endogenous TGF-beta1 released from AECs was observed. At time points 72 h, TGF-beta1 release mounted to 3451 pg/ml, which was much enough to induce EMT in vitro. These results demonstrated that AECs, under stimulation of TGF-beta1, underwent a conversion process into myofibroblasts in vitro.     

胰岛素样生长因子-1 对脂肪间充质干细胞增殖的影响

目的：探讨胰岛素样生长因子-1(insulin-like growth factor-1,IGF-1)对脂肪间充质干细胞(adipose-derived stem cells,ADSCs) 增殖的影响。方法：采用密度梯度离心法结合贴壁法分离脂肪间充质干细胞,接种于含体积分数为10%的胎牛血清的DMEM 培 养基中行贴壁培养。流式细胞仪检测ADSCs表面标志物(CD90、CD29、CD31、CD34、CD45)的表达情况,利用成骨、成脂诱导液诱 导ADSCs 向成骨细胞、成脂细胞分化,用碱性磷酸酶、油红O 染色观察。采用终浓度为0、5、10、15、20、30 ng/mL IGF的培养基培 养ADSCs,利用Edu 染色标记ADSCs,分析不同浓度的IGF-1 对ADSCs增殖的影响。结果：流式细胞术显示ADSCs的表型分子 CD90、CD29 呈阳性,CD31、CD45 呈阴性,成骨诱导后碱性磷酸酶染色阳性,成脂诱导后油红O染色可见大量脂滴,表明培养的 ADSCs具有成骨、成脂分化的能力。IGF-1 促进ADSCs 增殖的作用随IGF-1 的作用浓度的增加而增加,并逐渐趋于饱和,在趋于 15 滋g/mL的浓度时达到最大促增殖作用,且随着IGF-1 作用时间的延长其促ADSCs 增殖的作用逐渐增强。结论：本实验成功分 离培养ADSCs,IGF-1 对体外培养的ADSCs 有促进增殖的作用。     

Pravastatin up-regulates transforming growth factor-beta1 in THP-1 human macrophages: effect on scavenger receptor class A expression

Statins have been shown to interact with several monocyte/macrophage functions. We tested the effect of pravastatin on transforming growth factor-beta1 (TGF-beta1) production and its possible involvement in scavenger receptors class A (SRA) expression in human THP-1 cells. TGF-beta1s biological activity in THP-1 cell conditioned medium, evaluated by luciferase activity of transfected cell with a TGF-beta responsive promoter, was increased in a dose-dependent manner after incubation with pravastatin (1-20 microM). Pravastatin (1-20 microM) induced a dose-dependent increase in TGF-beta1 mRNA expression and protein production in THP-1 cells. PMA-induced SRA gene and protein expression was suppressed by pravastatin with a mean 3-fold decrease at 10 microM. This last effect was reversed by a mouse monoclonal anti-TGF-beta1 neutralizing antibody. PD98059, a specific inhibitor of MAP kinase cascade, completely reversed pravastatin-induced SRA down-regulation. p44 and p42 isoforms showed a dose-dependent phosphorylation after treatment with pravastatin (1-20 microM) which was inhibited by a mouse monoclonal anti-TGF-beta1 antibody. Our results demonstrate that pravastatin significantly up-regulates TGF-beta1 expression which may be in involved in down-regulation of SRA expression in THP-1 cell cultures. A new pathway for pravastatin effects in atherogenesis can be suggested.     

Collagen XI chain misassembly in cartilage of the chondrodysplasia (cho) mouse

Molecular mechanisms controlling the assembly of cartilage-specific types II, IX and XI collagens into a heteropolymeric network of uniformly thin, unbanded fibrils are not well understood, but collagen XI has been implicated. The present study on cartilage from the homozygous chondrodysplasia (cho/cho) mouse adds support to this concept. In the absence of alpha1(XI) collagen chains, thick, banded collagen fibrils are formed in the extracellular matrix of cho/cho cartilage. A functional knock-out of the type XI collagen molecule has been assumed. We have re-examined this at the protein level to see if, rather than a complete knock-out, alternative type XI chain assemblies were formed. Mass spectrometry of purified triple-helical collagen from the rib cartilage of cho/cho mice identified alpha1(V) and alpha2(XI) chains. These chains were recovered in roughly equal amounts based on Coomassie Blue staining of SDS-PAGE gels, in addition to alpha1(II)/alpha3(XI) collagen chains. Using telopeptide-specific antibodies and Western blot analysis, it was further shown that type V/XI trimers were present in the matrix cross-linked to each other and to type II collagen molecules to form heteropolymers. Cartilage from heterozygous (cho/+) mice contained a mix of alpha1(V) and alpha1(XI) chains and a mix of thin and thick fibrils on transmission electron microscopy. In summary, the results imply that native type XI collagen molecules containing an alpha1(XI) chain are required to form uniformly thin fibrils and support a role for type XI collagen as the template for the characteristic type II collagen fibril network of developing cartilage.     

Astrocytes express type VIII collagen during the repair process of brain cold injury

We recognized for the first time upregulation of type VIII collagen gene expression during the repair process in the mouse brain cold injury model. Immunohistochemical staining showed that type VIII collagen expression was around the necrotic region, where reactive astrocytes are frequently observed. Cultured astrocytes demonstrated a high expression of type VIII collagen genes. TGF-beta1 enhanced the expression of both alpha1(VIII) and alpha2(VIII) genes by astrocytes in culture. Further, we tested selected biological activities of type VIII collagen, compared with those of type I, IV, and V collagens and fibronectin. Astrocytes adhered to type VIII collagen via receptors requiring metal ions. Astrocyte migration on type VIII collagen was more stimulated than that observed on the other ECM molecules. These data indicate that type VIII collagen plays an important role in glial scar formation during the repair process by astrocytes.