摘要
目的探讨钙敏感受体(calcium sensing receptor,CaSR)/自噬信号轴在枸橼酸二乙酯(diethyl citrate,Et 2Cit)抑制慢性肾脏病(chronic kidney disease,CKD)血管钙化中的作用。方法将大鼠和血管平滑肌细胞(vascular smooth muscle cells,VSMCs)分别分为正常对照组、模型组及低、高剂量Et 2Cit组和高剂量Et 2Cit+NPS-2143(CaSR抑制剂)组进行干预,试剂盒法检测各组大鼠主动脉钙含量;茜素红染色法检测各组细胞钙化情况;qRT-PCR法和Western blotting检测各组大鼠主动脉中钙化相关蛋白、CaSR和自噬相关蛋白的mRNA和蛋白表达。结果与对照组相比,模型组钙化显著增加,Et 2Cit干预呈剂量依赖性地降低钙化程度(P<0.05);高剂量Et 2Cit+NPS-2143组钙含量较高剂量Et 2Cit组显著增加(P<0.05);与对照组相比,模型组平滑肌22α(smooth muscle 22α,SM22α)、CaSR和Beclin1的mRNA和蛋白表达降低(P<0.05),核心结合因子(runt related transcription factor 2,RUNX2)和P62表达升高(P<0.05);Et 2Cit干预能逆转以上变化(P<0.05);与高剂量Et 2Cit组相比,高剂量Et 2Cit+NPS-2143组的SM22α、CaSR和Beclin1显著降低,而RUNX2和P62显著增加(P<0.05)。结论Et 2Cit抑制CKD血管钙化,这一过程依赖CaSR和CaSR/自噬信号轴。
Objective To investigate the effects of calcium sensitive receptor(CaSR)/autophagy signaling axis on the inhibition of vascular calcification in chronic kidney disease(CKD)by diethyl citrate(Et 2Cit).Methods Rats and vascular smooth muscle cells(VSMCs)were divided into 5 groups:normal control group,model group,low-dose Et 2Cit group,high-dose Et 2Cit group and high-dose Et 2Cit+NPS-2143(CaSR inhibitor)group.After the intervention,the content of aorta calcium in each group was detected.Alizarin red staining was used to detect calcification in each cell group.mRNA expressions of calcification-related proteins,CaSR and autophagy-related proteins in the aorta of each group were detected by qRT-PCR.The expressions of the above proteins in each group were detected by Western blotting.Results Compared with the control group,the calcification in model group was significantly increased,and Et 2Cit intervention could reduce the calcification in a dose-dependent manner(P<0.05).Compared with high-dose Et 2Cit group,high-dose Et 2Cit+NPS-2143 group had significantly higher calcium content(P<0.05).Compared with those in the control group,the expressions of smooth muscle 22α(SM22α),CaSR and Beclin1 were decreased(P<0.05),while the expressions of runt related transcription factor 2(RUNX2)and P62 were increased(P<0.05).Et 2Cit intervention could reverse the above changes ( P <0.05). Compared with the high-dose Et 2Cit group, the high-dose Et 2Cit + NPS-2143 group had significantly lower SM22α, CaSR and Beclin1, and significantly higher RUNX2 and P62 levels ( P <0.05). Conclusion Et 2Cit inhibits CKD vascular calcification partly via the CaSR and CaSR/autophagy signal axis.
作者
杨艳艳
韩锦
桂保松
姚智会
YANG Yanyan;HAN Jin;GUI Baosong;YAO Zhihui(Department of Nephrology,Xi'an 710004,China;Department of Cardiology,The Second Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710004,China)
出处
《西安交通大学学报（医学版）》
CAS
CSCD
北大核心
2020年第5期701-705,共5页
Journal of Xi’an Jiaotong University（Medical Sciences）
基金
国家自然科学基金资助项目(No.81570673),美捷登青年科学家研究基金(No.MJR20201127)。
作者简介
通信作者:姚智会,助理研究员.E-mail:310262382@sina.com。
