SP对NE诱导的体外培养大鼠心肌细胞凋亡的影响及机制  

Effect of substance P on norepinephrine-induced apoptosis in cultured rat cardiomyocytes and its mechanisms

作　　者：马艳霞[1,2] 孙韬[1,2] 郭政[1,2] 

机构地区：[1]山西医科大学麻醉学系,太原030001 [2]山西医科大学第二临床医学院麻醉科

出　　处：《山西医科大学学报》2015年第4期289-292,共4页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(30772083;30972860);山西医科大学博士启动基金资助项目(03201425)

摘　　要：目的观察P物质(SP)对去甲肾上腺素(NE)诱导的体外培养大鼠心肌细胞凋亡的影响及机制。方法选用出生1-3 d的SD大鼠建立新生大鼠心肌细胞体外培养模型,将12孔培养72 h的心肌细胞随机分为对照组(不加任何干预药物)、NE组、SP+NE组、D-SP(SP的特异性受体拮抗剂)+SP+NE组,每组n=3。以上各组中,NE、SP、D-SP终浓度分别为10-5,10-7,10-6mol/L。药物作用3 h后采用流式细胞术(FCM)测定凋亡率,采用Western blot的方法检测αB晶体蛋白(CRYAB)的含量变化,采用实时荧光定量PCR(RT-PCR)的方法检测CRYAB mRNA的变化。结果与对照组比较,NE组凋亡率升高,CRYAB蛋白表达和mRNA相对量均下降(P<0.05);与NE组比较,SP+NE组凋亡率下降,CRYAB蛋白表达与mRNA升高(P<0.05);与SP+NE组比较,D-SP+SP+NE组凋亡率升高,CRYAB蛋白表达与mRNA均下降(均P<0.05)。结论SP抑制NE所诱导的体外培养大鼠心肌细胞凋亡,该作用由特异性受体介导,与调控CRYAB表达实现表达水平内稳态有关。Objective To explore the effect of substance P（ SP） on the apoptosis induced by norepinepherine（ NE） in cultured neonatal rat cardiomyocytes and its mechanisms.Methods The neonatal cardiomyocytes from the 1-3 d-old Sprague-Dawley rats were isolated and cultured.The cardiomyocytes incubated for 72 h were used for further experiments.Twelve wells of rat cardiomyocytes were randomly divided into 4 groups（ 3 wells in each group） ：control group,NE group,SP ＋ NE group,D-SP（ a special SP receptor antagonist）＋ SP ＋ NE group.The final concentrations of NE,SP,D-SP were 10-5,10-7,10-6mol / L in each group,respectively.The apoptosis rate of the cultured cardiomyocytes was examined by FCM at 3 h after incubation with NE.The expression of αB-crystallin protein was analyzed by Western blot,and the coding mRNA of αB-crystallin was detected by quantitative real-time PCR（ qRT-PCR）.Results The apoptosis rate of the cardiomyocytes was significantly higher in NE group than that in control group by FCM（ P 〈0.05）.However,compared with control group,the expression and coding mRNA level of αB-crystallin were markedly decreased in NE group（ P 〈0.05）.The apoptosis rate was lower,and the protein expression and coding mRNA of αB-crystallin were higher in SP ＋ NE group than that in NE group（ all P 〈0.05）.D-SP,a specific antagonist of neurokinin 1 receptor,reversed the effect of SP.Conclusion SP inhibits NE-induced apoptosis in cultured cardiomyocytes,which might be associated with modulating activity of genes coding αB-crystallin and maintaining homeostasis of the small heat shock protein via specific neurokinin 1 receptor.

关 键 词：凋亡 心肌细胞 P物质 去甲肾上腺素 αB晶体蛋白 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]