纤维细胞生长因子21对血管钙化大鼠特异性caspase-12凋亡途径作用的研究  

The protective effects of fibroblast growth factor 21 on vascular calcification via inhibiting specific caspase-12 mediated apoptosis pathway

作　　者：史雨晨[1] 柳景华[1] SHI Yuchen;LIU Jinghua(Department of Cardiology,Beijing Anzhen Hospital,Capital Medical University,Beijing Institute of Heart,Lung and Blood Vessel Diseases,Beijing 100029,China)

机构地区：[1]首都医科大学附属北京安贞医院-北京市心肺血管疾病研究所心内科,100029

出　　处：《心肺血管病杂志》2021年第6期628-633,共6页Journal of Cardiovascular and Pulmonary Diseases

基　　金：国家自然科学基金(81970291);国家重点基础研究发展计划(973计划)课题(2015CB554404)。

摘　　要：目的:探究纤维细胞生长因子21(FGF21)对大鼠血管钙化的保护作用及相关机制;为深入研究血管钙化机制,研发预测因子及治疗药物,提供基础依据。方法:雄性成年SD大鼠,21只,平均体质量(190±10)g,按电脑随机数字表法平均分为对照组、血管钙化组和血管钙化+FGF21组,每组7只。血管钙化组采用维生素D3联合尼古丁(VDN)诱导大鼠血管钙化模型。以碱性磷酸酶(ALP)试剂盒和钙含量试剂盒检测大鼠主动脉组织ALP活性和钙含量,以TUNEL法检测大鼠主动脉细胞凋亡情况,以免疫荧光法检测大鼠主动脉细胞α-actin表达情况,以免疫组织化学法检测大鼠主动脉细胞caspase-3及caspase-12表达情况。结果:血管钙化组大鼠主动脉钙含量和ALP活性相较对照组分别升高6.84倍(P<0.01)和3.07倍(P<0.05)。而与单纯血管钙化组大鼠相比,FGF21可显著降低大鼠主动脉钙含量和ALP活性57.9%(P<0.05)和49.3%(P<0.05)。血管钙化组大鼠主动脉中凋亡细胞数目显著增加,而α-actin表达量显著降低。给予血管钙化大鼠FGF21后,凋亡细胞数目减少,同时α-actin表达量有所增加。进一步应用免疫组织化学法观察显示,血管钙化组大鼠主动脉细胞caspase-3及caspase-12表达量显著增加,而FGF21可以抑制两者的表达。结论:外源性FGF21对大鼠血管钙化具有保护作用,其机制可能部分与抑制内质网应激介导的特异性caspase-12凋亡通路信号有关。Objective: The present study aimed to investigate the possible role of FGF21 on vascular calcification via inhibiting specific caspase-12 mediated apoptosis pathway. Methods: The male SD rats(180-200 mg) were divided into three groups:①control group,②vitamin D3 plus nicotine(VDN) group and ③ FGF21 treated VDN group. After 4 weeks treatment, the aortic tissues were homogenized for calcium content and alkaline phosphatases(ALP) activity. The apoptosis of vascular smooth muscle cells(VSMCs) were tested by TUNEL staining. And the VSMCs expression of α-actin were tested by immunofluorescence staining. The proteins expression of caspase-3 and caspase-12 in aortic tissue were tested by immunohistochemistry. Results: The vascular calcification model induced by VDN was successfully and stably. With 28 days subcutaneous administration of FGF21, the calcium content and ALP activity in the aortas were decreased by 57.9%(P<0.05)and 49.3%(P<0.05). Furthermore, the apoptotic cells were increased by TUNEL staining associating with the decreased expression of α-actin by immunofluorescence staining as compared with the control group. And FGF21 treatment can reverse these tendencies. In addition, compared with the control group, the proteins expression of caspase-3 and caspase-12 in aortic tissue were increased by immunohistochemistry. And under the treatment of FGF21, both of them were decreased as compared with the VDN group. Conclusions: In our research, we provide the first evidence that exogenous FGF21 treatment can alleviate the VDN induced vascular calcification at least in part via suppressing specific caspase-12 mediated apoptosis pathway in rats.

关 键 词：成纤维细胞生长因子21 血管钙化 凋亡 

分 类 号：R54[医药卫生—心血管疾病]