Apelin-13通过e NOS/NO促进糖尿病小鼠主动脉细胞焦亡相关蛋白的表达

目的探讨血管活性多肽apelin-13是否通过eNOS/NO途径影响糖尿病小鼠主动脉细胞焦亡相关蛋白的表达。方法以8周龄C57/BL小鼠作为对照组小鼠;以8周龄kkAy小鼠作为2型糖尿病小鼠模型。给糖尿病小鼠皮下埋植缓释泵,持续灌注apelin-13处理14 d,剂量为30μg/(kg·d)或apelin-13和L-NAMENG-nitro-L-arginine methyl ester,L-NAME,eNOS抑制剂,剂量为10 mg/(kg·d)]共同处理14 d。用无创尾压法检测各组小鼠处理前后的血压值;取血检测血糖和糖化血红蛋白;取腹主动脉,进行HE染色,显微镜下观察血管形态学改变,免疫组织化学染色分析小鼠主动脉内eNOS和细胞焦亡相关信号蛋白NLRP3、caspase-1、gasdermin D的表达水平。结果与对照组相比,糖尿病组小鼠主动脉中eNOS含量明显升高(P<0.05),焦亡相关信号蛋白NLRP3、caspase-1、gasdermin D的表达水平升高,经过apelin-13处理后,糖尿病小鼠血管壁eNOS含量进一步升高(P<0.05),焦亡相关信号蛋白NLRP3、caspase-1、gasdermin D的表达水平也随之升高(P<0.05),给予eNOS抑制剂L-NAME之后,糖尿病小鼠主动脉eNOS含量降低(P<0.05),焦亡相关信号蛋白NLRP3、caspase-1、gasdermin D的表达水平也随之降低(P<0.05)。结论Apelin-13可能通过eNOS/NO途径促进糖尿病小鼠主动脉细胞焦亡相关蛋白的表达,导致糖尿病小鼠血管结构损伤和功能。

Apelin-13 promotes expression of pyroptosis related protein in aortic cells in diabetic mice through eNOS/NO pathway

Objective To observe the effects and mechanisms of Apelin-13 on expression of pyroptosis related proteins in aorta of diabetic mice.Methods C57/BL mice of eight weeks old were used as control group;kkAy mice of eight weeks old were used as type 2 diabetic models;osmotic pumps were used to treat kkAy mice with apelin-13 at a rate of 30μg/(kg·d),and L-NAME(eNOS inhibitor)was injected intraperitoneally at a dose of 10 mg/(kg·d)to kkAy mice.Blood was collected for detection of Hb1Ac.The aortae were harvested and fixed.Morphological changes were observed with HE staining.Expression of eNOS,NLRP3,caspase-1 and gasdermin D were measured with immunohistochemical staining.Results Compared to the control group,the level of eNOS in the aorta of diabetic mice was significantly higher than that in control mice(P<0.05),the levels of NLRP3,caspase-1 and gasdermin D were also higher than that in control mice.After apelin-13 treatment,the expressions of eNOS,NLRP3,caspase-1 and gasdermin D were further increased(P<0.05).After L-NAME and apelin-13 treatment,the expressions of eNOS,NLRP3,caspase-1 and gasdermin D were reduced as compared to apelin-13 treatment alone(P<0.05).Conclusions Apelin-13 may promote the expression of pyroptosis related protein in aortic cells by increasing eNOS/NO pathway,which would induce structural and functional damage in diabetic arteries.