Background The present study aimed to investigate the detailed mode and specific sites for their binding as well as the functional relevance of this binding in the phenotypic proliferation of vascular smooth muscle ce...Background The present study aimed to investigate the detailed mode and specific sites for their binding as well as the functional relevance of this binding in the phenotypic proliferation of vascular smooth muscle cells(SMCs). Methods CREG knocked-down SMCs were employed to evaluate the biological activity of wtCREG and mCREG.Expressions of SMC differentiation markers SM myosin heavy chain(SM-MHC),SM-actin,heavy caldesmon and myocardin were determined by Western blotting using specific antibodies. Cellular growth of SMCs was assessed by bromide dewuridine (BrdU) incorporation and cell cycle analysis on fluorescence-activated cell sorting(FACS).A solid-phase binding assay was used to study the binding of CREG to extracellular domains of M6P/IGF2R.The cellular co-localization of the two recombinant CREGs with M6P/IGF2R was detected on SMC surface by immunoprecipitation and immunofluorescence analysis.Results The molecular weight of wtCREG was around 30 kD while that of the mCREG was～25 kD.Treatment of wtCREG with PNGase F reduced its molecular weight from～30 kD to～25 kD,whereas PNGase F treatment had no effect on the molecular weight of mCREG.Both wtCREG and mCREG proteins enhanced SMC differentiation,inhibited BrdU incorporation,and arrested cell cycle progression when added to the culture medium.In CREG knocked-down SMCs,the amount of CREG detected by immunoblotting in M6P/IGF2R immunoprecipitates was significantly reduced when compared to normal cells.Both recombinant CREGs co-immunoprecipitated with M6P/IGF2R, although slightly reduced amount of the mutant CREG was detected in M6P/IGF2R immunoprecipitates.Immunostaining revealed that His-tagged CREGs co-localized with IGF2R on the cell surface in a glycosylation-independent manner.In vitro binding assay showed that CREGs bound to M6P/ IGF2R extracellular domains 7-10 and 11-13 in a glycosylation -dependent and -independent manner,respectively.Further blocking experiments using soluble M6P/IGF2R fragments and M6P/IGF2R neutralizing antibody indicated that the biological activities of recombinant CREGs in SMC growth and the up-regulation of SMC differentiation markers were all abolished by treatment with the M6P/IGF2R neutralizing antibody. However,although the growth inhibitory effect of wtCREG was nearly abolished by D7-10 or D11-13,the effect of mCREG was only reversed by Dll-13,indicating that the binding to domains 11-13 is required for CREG to modulate the proliferation of SMCs.Conclusions These data suggest that solubleCREG proteins can exert their biological function via binding to the extracellular domains 7-10 and 11-13 of cell surface M6P/IGF2R in both a glycosylation-dependent and -independent manner.展开更多
目的探讨胰高血糖素样肽-1(GLP-1)激动剂利拉鲁肽对2型糖尿病(T2DM)合并动脉粥样硬化患者血管重构、肠道菌群代谢物及血清C1q/肿瘤坏死因子相关蛋白13(CTRP13)的影响。方法选择2021年6月至2024年6月于该院接受治疗的214例T2DM合并动脉...目的探讨胰高血糖素样肽-1(GLP-1)激动剂利拉鲁肽对2型糖尿病(T2DM)合并动脉粥样硬化患者血管重构、肠道菌群代谢物及血清C1q/肿瘤坏死因子相关蛋白13(CTRP13)的影响。方法选择2021年6月至2024年6月于该院接受治疗的214例T2DM合并动脉粥样硬化患者,采用随机数字表法分为观察组与对照组,每组107例。对照组在常规治疗基础上给予二甲双胍治疗,观察组在对照组基础上给予利拉鲁肽治疗。比较2组患者治疗前后血糖[餐后2小时血糖(2 h PG)、空腹血糖(FPG)、糖化血红蛋白(HbA1c)]、血脂[高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、甘油三酯(TG)、总胆固醇(TC)]、血压[舒张压(DBP)、收缩压(SBP)]、血管重构指标、肠道菌群代谢物[三甲胺氧化物(TMAO)]、炎症因子[C反应蛋白(CRP)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)]及CTRP13水平,以及临床疗效和不良反应情况。结果观察组总有效率为87.9%,高于对照组的65.4%(P<0.05)。治疗后2组血糖、血压、LDL-C、TG、TC水平明显下降,且观察组低于对照组,治疗后2组HDL-C水平明显升高,且观察组HDL-C水平高于对照组,差异均有统计学意义(P<0.05)。与治疗前相比,2组患者CTRP13水平均升高,而TMAO水平均降低,且观察组CTRP13水平高于对照组,TMAO水平低于对照组,差异均有统计学意义(P<0.05)。与治疗前相比,2组患者治疗后CRP、IL-6、TNF-α水平降低(P<0.05),且观察组低于对照组(P<0.05)。治疗后观察组颈内动脉内膜中层厚度(IMT)、不稳定斑块检出率低于对照组(P<0.05)。观察组治疗前后血管正性重构发生率及负性重塑发生率比较,差异均无统计学意义(P>0.05)。对照组、观察组不良反应发生率分别为14.0%、16.8%,差异无统计学意义(χ^(2)=0.322,P=0.570)。结论利拉鲁肽有利于控制T2DM合并动脉粥样硬化患者血糖、血脂、血压水平,减少TMAO释放,提升CTRP13水平,减轻机体炎症反应,改善血管重构,降低IMT,稳定斑块,且安全性较好。展开更多
目的:探究子痫前期患者血清胎盘蛋白-13(Placental protein 13,PP13)、基质金属蛋白酶抑制因子2(Tissue inhibitor of metalloproteinase,TIMP-2)、缺氧诱导因子-1α(Hypoxia inducible factor-1α,HIF-1α)水平与胎儿生长受限的相关性...目的:探究子痫前期患者血清胎盘蛋白-13(Placental protein 13,PP13)、基质金属蛋白酶抑制因子2(Tissue inhibitor of metalloproteinase,TIMP-2)、缺氧诱导因子-1α(Hypoxia inducible factor-1α,HIF-1α)水平与胎儿生长受限的相关性及临床意义。方法:选取2021年9月至2023年9月期间本院收治的94例子痫前期孕妇作为研究对象。根据孕妇分娩后胎儿的体重情况评估胎儿是否发生胎儿生长受限(Fetal growth restriction,FGR),将研究对象分为FGR组(n=31)和非FGR组(n=63)。另选取同期内的94例健康孕妇作为研究对象的对照组。对比三组血清PP13、TIMP-2、HIF-1α水平。分析血清PP13、TIMP-2、HIF-1α水平与FGR的相关性和独立影响因素。分析各指标对子痫前期患者FGR发生的预测价值。结果:FGR组的血清PP13水平明显低于非FGR组和对照组,非FGR组血清PP13水平低于明显对照组(P<0.05)。FGR组的血清TIMP-2和HIF-1α水平明显高于非FGR组和对照组,非FGR组的血清TIMP-2和HIF-1α水平明显高于对照组(P<0.05)。血清PP13水平与FGR发生呈负相关,血清TIMP-2、HIF-1α水平与FGR发生呈正相关(P<0.05)。血清PP13是FGR发生的独立保护因素,血清TIMP-2、HIF-1α是FGR发生的独立危险因素(P<0.05)。血清PP13、TIMP-2、HIF-1α水平联合预测FGR发生的曲线下面积(Area under the curve,AUC)明显大于各血清指标的单独预测(P<0.05)。结论:血清PP13、TIMP-2、HIF-1α水平与子痫前期患者FGR发生关系密切,各指标联合检测可为预测FGR发生提供有效参考。展开更多
目的 探讨血清孤独G蛋白偶联受体配体-13(solitary G protein-coupled receptor ligand-13,Apelin-13)、碱性成纤维细胞生长因子(fibroblast growth factor 2,FGF2)、中枢神经特异蛋白(central nervous system specific protein B,S100B...目的 探讨血清孤独G蛋白偶联受体配体-13(solitary G protein-coupled receptor ligand-13,Apelin-13)、碱性成纤维细胞生长因子(fibroblast growth factor 2,FGF2)、中枢神经特异蛋白(central nervous system specific protein B,S100B)水平与脑卒中后抑郁(post-stroke depression, PSD)的关系。方法 回顾性分析河南科技大学附属黄河医院/黄河三门峡医院2022年4月至2024年1月收治的260例脑卒中患者临床资料,随访1个月,参考17项汉密尔顿抑郁量表(Hamilton Depression Rating Scale, HAMD-17)评估结果统计PSD发生率,并将其分为发生组和未发生组,比较两组血清Apelin-13、FGF2、S100B水平及其他临床资料,采用Logistic回归模型分析PSD发生的影响因素;绘制受试者工作特征曲线(Receiver operating characteristic curve, ROC)评价血清Apelin-13、FGF2、S100B对PSD的预测价值。结果 PSD发生率为37.31%;与未发生组比较,发生组女性、糖尿病、左侧大脑半球病灶占比、美国国立卫生研究院卒中量表(national Institutes of health stroke scale, NIHSS)评分、同型半胱氨酸水平、系统性免疫炎症指数(systemic immune-inflammatory index, SII)、血清Apelin-13、S100B水平较高(P<0.05),月均可支配医疗费用、血清FGF2水平较低(P<0.05);月均可支配医疗费用(OR=0.732,95%CI:0.599~0.894)、左侧大脑半球病灶(OR=1.478,95%CI:1.159~1.885)、NIHSS评分(OR=1.293,95%CI:1.095~1.527)、同型半胱氨酸水平(OR=1.278,95%CI:1.058~1.542)、SII(OR=1.370,95%CI:1.133~1.657)、Apelin-13水平(OR=1.278,95%CI:1.086~1.503)、FGF2水平(OR=0.807,95%CI:0.689~0.946)、S100B水平(OR=1.357,95%CI:1.089~1.690)为PSD发生的影响因素(P<0.05);血清Apelin-13、FGF2、S100B联合预测PSD的灵敏度高于单独预测,曲线下面积(area under curve, AUC)高于单独预测方法(P<0.05),特异度与单独预测基本一致。结论 血清Apelin-13、FGF2、S100B水平与月均可支配医疗费用、左侧大脑半球病灶、NIHSS评分、同型半胱氨酸、SII可能是PSD发生的影响因素,且血清Apelin-13、FGF2、S100B可能对PSD有预测价值,其中联合预测价值更显著。展开更多
文摘Background The present study aimed to investigate the detailed mode and specific sites for their binding as well as the functional relevance of this binding in the phenotypic proliferation of vascular smooth muscle cells(SMCs). Methods CREG knocked-down SMCs were employed to evaluate the biological activity of wtCREG and mCREG.Expressions of SMC differentiation markers SM myosin heavy chain(SM-MHC),SM-actin,heavy caldesmon and myocardin were determined by Western blotting using specific antibodies. Cellular growth of SMCs was assessed by bromide dewuridine (BrdU) incorporation and cell cycle analysis on fluorescence-activated cell sorting(FACS).A solid-phase binding assay was used to study the binding of CREG to extracellular domains of M6P/IGF2R.The cellular co-localization of the two recombinant CREGs with M6P/IGF2R was detected on SMC surface by immunoprecipitation and immunofluorescence analysis.Results The molecular weight of wtCREG was around 30 kD while that of the mCREG was～25 kD.Treatment of wtCREG with PNGase F reduced its molecular weight from～30 kD to～25 kD,whereas PNGase F treatment had no effect on the molecular weight of mCREG.Both wtCREG and mCREG proteins enhanced SMC differentiation,inhibited BrdU incorporation,and arrested cell cycle progression when added to the culture medium.In CREG knocked-down SMCs,the amount of CREG detected by immunoblotting in M6P/IGF2R immunoprecipitates was significantly reduced when compared to normal cells.Both recombinant CREGs co-immunoprecipitated with M6P/IGF2R, although slightly reduced amount of the mutant CREG was detected in M6P/IGF2R immunoprecipitates.Immunostaining revealed that His-tagged CREGs co-localized with IGF2R on the cell surface in a glycosylation-independent manner.In vitro binding assay showed that CREGs bound to M6P/ IGF2R extracellular domains 7-10 and 11-13 in a glycosylation -dependent and -independent manner,respectively.Further blocking experiments using soluble M6P/IGF2R fragments and M6P/IGF2R neutralizing antibody indicated that the biological activities of recombinant CREGs in SMC growth and the up-regulation of SMC differentiation markers were all abolished by treatment with the M6P/IGF2R neutralizing antibody. However,although the growth inhibitory effect of wtCREG was nearly abolished by D7-10 or D11-13,the effect of mCREG was only reversed by Dll-13,indicating that the binding to domains 11-13 is required for CREG to modulate the proliferation of SMCs.Conclusions These data suggest that solubleCREG proteins can exert their biological function via binding to the extracellular domains 7-10 and 11-13 of cell surface M6P/IGF2R in both a glycosylation-dependent and -independent manner.
文摘目的探讨胰高血糖素样肽-1(GLP-1)激动剂利拉鲁肽对2型糖尿病(T2DM)合并动脉粥样硬化患者血管重构、肠道菌群代谢物及血清C1q/肿瘤坏死因子相关蛋白13(CTRP13)的影响。方法选择2021年6月至2024年6月于该院接受治疗的214例T2DM合并动脉粥样硬化患者,采用随机数字表法分为观察组与对照组,每组107例。对照组在常规治疗基础上给予二甲双胍治疗,观察组在对照组基础上给予利拉鲁肽治疗。比较2组患者治疗前后血糖[餐后2小时血糖(2 h PG)、空腹血糖(FPG)、糖化血红蛋白(HbA1c)]、血脂[高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、甘油三酯(TG)、总胆固醇(TC)]、血压[舒张压(DBP)、收缩压(SBP)]、血管重构指标、肠道菌群代谢物[三甲胺氧化物(TMAO)]、炎症因子[C反应蛋白(CRP)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)]及CTRP13水平,以及临床疗效和不良反应情况。结果观察组总有效率为87.9%,高于对照组的65.4%(P<0.05)。治疗后2组血糖、血压、LDL-C、TG、TC水平明显下降,且观察组低于对照组,治疗后2组HDL-C水平明显升高,且观察组HDL-C水平高于对照组,差异均有统计学意义(P<0.05)。与治疗前相比,2组患者CTRP13水平均升高,而TMAO水平均降低,且观察组CTRP13水平高于对照组,TMAO水平低于对照组,差异均有统计学意义(P<0.05)。与治疗前相比,2组患者治疗后CRP、IL-6、TNF-α水平降低(P<0.05),且观察组低于对照组(P<0.05)。治疗后观察组颈内动脉内膜中层厚度(IMT)、不稳定斑块检出率低于对照组(P<0.05)。观察组治疗前后血管正性重构发生率及负性重塑发生率比较,差异均无统计学意义(P>0.05)。对照组、观察组不良反应发生率分别为14.0%、16.8%,差异无统计学意义(χ^(2)=0.322,P=0.570)。结论利拉鲁肽有利于控制T2DM合并动脉粥样硬化患者血糖、血脂、血压水平,减少TMAO释放,提升CTRP13水平,减轻机体炎症反应,改善血管重构,降低IMT,稳定斑块,且安全性较好。
文摘目的:探究子痫前期患者血清胎盘蛋白-13(Placental protein 13,PP13)、基质金属蛋白酶抑制因子2(Tissue inhibitor of metalloproteinase,TIMP-2)、缺氧诱导因子-1α(Hypoxia inducible factor-1α,HIF-1α)水平与胎儿生长受限的相关性及临床意义。方法:选取2021年9月至2023年9月期间本院收治的94例子痫前期孕妇作为研究对象。根据孕妇分娩后胎儿的体重情况评估胎儿是否发生胎儿生长受限(Fetal growth restriction,FGR),将研究对象分为FGR组(n=31)和非FGR组(n=63)。另选取同期内的94例健康孕妇作为研究对象的对照组。对比三组血清PP13、TIMP-2、HIF-1α水平。分析血清PP13、TIMP-2、HIF-1α水平与FGR的相关性和独立影响因素。分析各指标对子痫前期患者FGR发生的预测价值。结果:FGR组的血清PP13水平明显低于非FGR组和对照组,非FGR组血清PP13水平低于明显对照组(P<0.05)。FGR组的血清TIMP-2和HIF-1α水平明显高于非FGR组和对照组,非FGR组的血清TIMP-2和HIF-1α水平明显高于对照组(P<0.05)。血清PP13水平与FGR发生呈负相关,血清TIMP-2、HIF-1α水平与FGR发生呈正相关(P<0.05)。血清PP13是FGR发生的独立保护因素,血清TIMP-2、HIF-1α是FGR发生的独立危险因素(P<0.05)。血清PP13、TIMP-2、HIF-1α水平联合预测FGR发生的曲线下面积(Area under the curve,AUC)明显大于各血清指标的单独预测(P<0.05)。结论:血清PP13、TIMP-2、HIF-1α水平与子痫前期患者FGR发生关系密切,各指标联合检测可为预测FGR发生提供有效参考。
