雌二醇经雌激素受体调控分拣蛋白相关受体A对小鼠海马神经元树突棘密度和突触蛋白表达的影响

Effect of estradiol regulating sortilin-related receptor A expression on hippocampal spine density and synaptic protein expression via estrogen receptor of mouse

目的探讨雌二醇(E_(2))调控分拣蛋白相关受体A(SorLA)表达对小鼠海马神经元树突棘密度和突触蛋白表达的影响及机制。方法32只C57BL/6雌性小鼠随机分为假手术(sham)组、卵巢切除(OVX)组、卵巢切除+E_(2)组(OVX+E_(2))组、卵巢切除+氟维司琼(Fu)+E_(2)组(OVX+Fu+E_(2))组,通过Western blotting和免疫组织化学染色检测小鼠海马CA1区SorLA表达;40只C57BL/6雌性小鼠随机分为假手术+空白病毒(sham+NC-sgRNA)组、卵巢切除+空白病毒(OVX+NC-sgRNA)组、卵巢切除+空白病毒+E_(2)(OVX+NC-sgRNA+E_(2))组、卵巢切除+SorLA敲降病毒+E_(2)组(OVX+Sorl1-sgRNA+E_(2))组,通过高尔基染色检测小鼠海马CA1区神经元树突棘密度;通过Western blotting和免疫组织化学染色检测小鼠海马CA1区突触后致密蛋白95(PSD-95)和突触蛋白1(SYN1)表达。结果与sham组相比,和OVX组和OVX+Fu+E_(2)组SorLA蛋白表达显著降低(P<0.05),和OVX+E_(2)组差异无显著性(P>0.05);与sham+NC-sgRNA组相比,OVX+NC-sgRNA组和OVX+Sorl1-sgRNA+E_(2)组树突棘密度和PSD-95、SYN1表达显著降低(P<0.05),与OVX+NC-sgRNA+E_(2)组差异无显著性(P>0.05)。结论E_(2)可通过雌激素受体上调SorLA表达,进而增加小鼠海马神经元树突棘密度和突触蛋白表达。

Objective To study the effect and mechanism of estradiol(E_(2))regulating sortilin-related receptor A(SorLA)expression on hippocampal spine density and synaptic protein expression of mouse.Methods Thirty-two C57BL/6 female mice were randomly divided into sham group,ovariectomized(OVX)group,OVX+E_(2)group,and OVX+fulvestrant(Fu)+E_(2)group;The SorLA expression in hippocampal CAl region of mouse was detected by Western blotting and immunohistochemical staining.Forty C57BL/6 female mice were randomly divided into sham+NC-sgRNA group,OVX+NC-sgRNA group,OVX+NC-sgRNA+E_(2)group,and OVX+sortilin-related receptor 1(Sorll)-sgRNA+E_(2);the spine density in hippocampal CA1 region was detected by Golgi-Cox staining and the postsynaptic density protein 95(PSD-95)and synapsin-1(SYN1)expression in hippocampal CA1 region were detected by Western blotting and immunohistochemical staining.Results Compared with the sham group,the SorLA expression in OVX group and OVX+Fu+E_(2)group decreased significantly(P<0.05),but OVX+E_(2) group had no significant difference(P>0.05).Compared with the sham+NC-sgRNA group,the density of dendritic spines,and the PSD-95,SYN1 expression of OVX+NC-sgRNA group and OVX+SorllsgRNA+E_(2)group decreased significantly(P<0.05),but OVX+NC-sgRNA+E_(2) group had no significant difference(P>0.05).Conclusion E_(2) may upregulate SorLA expression through estrogen receptor and then promote the hippocampal spine density and synaptic protein expression of mouse.