丹参素通过JAK/STAT信号通路对冠心病大鼠的心肌保护作用及对血栓弹力图的影响

Effect of Danshensu on Myocardial Protection of Coronary Heart Disease Rats through JAK/STAT Signal Pathway and Its Effect on Thromboelastography

目的研究丹参素通过蛋白酪氨酸激酶(JAK)/信号传导子和转录激活子(STAT)信号通路对冠心病模型大鼠的心肌保护作用以及对血栓弹力图(TEG)相关指标的影响。方法选取60只无特定病原体(SPF)级健康SD大鼠,随机抽取12只作为正常对照组,剩余48只饲喂高脂饮食和腹腔注射垂体后叶素建立冠心病模型大鼠。将45只建模成功的大鼠随机分为模型组(11只)、阳性对照组(11只)、丹参素低剂量组(11只)、丹参素高剂量组(12只)。丹参素低剂量组、丹参素高剂量组分别按10 mL/kg灌胃1 mg/mL、2 mg/mL的丹参素溶液,阳性对照组按10 mL/kg灌胃盐酸地尔硫[艹卓]溶液,正常对照组及模型组按10 mL/kg灌胃生理盐水,每日1次,连续2周。比较各组TEG相关指标、心肌损伤指标、氧化应激相关指标水平;原位缺口末端标记(TUNEL)法检测心肌细胞凋亡情况;比较心肌组织中蛋白酪氨酸激酶2(JAK2)、信号转导子和转录激活子1(STAT1)、信号转导子和转录激活子3(STAT3)mRNA水平及其磷酸化JAK2(p-JAK2)/JAK2、磷酸化STAT1(p-STAT1)/STAT1、磷酸化STAT3(p-STAT3)/STAT3比值。结果与正常对照组比较,模型组、丹参素低剂量组、丹参素高剂量组、阳性对照组R值缩短,Angle角及MA值增大,肌酸激酶同工酶(CK-MB)、心肌肌钙蛋白I(cTnI)、丙二醛(MDA)、心肌细胞凋亡指数、p-JAK2/JAK2、p-STAT1/STAT1、p-STAT3/STAT3均升高,超氧化物歧化酶(SOD)、总抗氧化能力(T-AOC)均降低,差异均有统计学意义(P<0.05);与模型组比较,丹参素低剂量组、丹参素高剂量组、阳性对照组R值延长,Angle角及MA值减小,CK-MB、cTnI、MDA、心肌细胞凋亡指数、p-JAK2/JAK2、p-STAT1/STAT1、p-STAT3/STAT3均降低,SOD、T-AOC均升高,差异均有统计学意义(P<0.05);与丹参素低剂量组比较,丹参素高剂量组、阳性对照组R值延长,Angle角和MA值减小,CK-MB、cTnI、MDA、心肌细胞凋亡指数、p-JAK2/JAK2、p-STAT1/STAT1、p-STAT3/STAT3均降低,SOD、T-AOC均升高,差异均有统计学意义(P<0.05);丹参素高剂量组较阳性对照组R值延长,Angle角及MA值减小,CK-MB、cTnI、MDA、心肌细胞凋亡指数、p-JAK2/JAK2、p-STAT1/STAT1、p-STAT3/STAT3降低,SOD、T-AOC升高,差异均有统计学意义(P<0.05)。各组JAK2、STAT1、STAT3 mRNA相对表达量比较,差异均无统计学意义(P>0.05)。结论丹参素可改善TEG相关指标及氧化应激,具有心肌保护作用,可能通过抑制JAK/STAT信号通路发挥调控作用。

Objective To study the protective effect of Danshensu on myocardium in coronary heart disease(CHD)model rats through Janus kinase(JAK)/signal transducer and activator of transcription(STAT)signal pathway and its effect on thromboelastography(TEG)related indicators.Methods Sixty healthy SPF-grade SD rats were selected,twelve randomly selected rats were used as normal control group,and the remaining 48 rats were fed high-fat diet and intraperitoneal injection of pituitary lutein to establish CHD model.Forty-five successfully modeled rats were randomly divided into model group(11 rats),positive control group(11 rats),Danshensu low-dose group(11 rats),and Danshensu high-dose group(12 rats).The rats in Danshensu low-dose group and Danshensu high-dose group were administrated with 1 mg/mL and 2 mg/mL of Danshensu solution according to 10 mL/kg body weight,respectively,and the rats in positive control group were administered with hydrochloric acid diltiazem solution according to 10 mL/kg body weight,the rats in normal control group and model group were administrated with physiological saline according to 10 mL/kg,once a day for 2 weeks.The levels of TEG-related indexes,myocardial injury indexes,and oxidative stress-related indexes were compared between groups.The apoptosis of cardiomyocytes was detected by in situ end labeling(TUNEL).Janus kinase 2(JAK2),signal transducer and activator of transcription(STAT)1,STAT3 mRNA levels,and phosphorylated JAK2(p-JAK2)/JAK2,phosphorylated STAT1(p-STAT1)/STAT1,phosphorylated STAT3(p-STAT3)/STAT3 ratio were compared.Results Compared with normal control group,R values were shorter,Angle angles and MA values were increased,creatine phosphokinase-MB(CK-MB),cardiac troponin I(cTnI),malondialdehyde(MDA),myocardial cell apoptosis index,p-JAK2/JAK2,p-STAT1/STAT1,p-STAT3/STAT3 were increased,and superoxide dismutase(SOD)and total antioxidant capacity(T-AOC)were decreased in the model group,Danshensu low-dose group,Danshensu high-dose group,positive control group(P<0.05).Compared with model group,R values were longer,Angle angles and MA values were decreased,CK-MB,cTnI,MDA,myocardial cell apoptosis index and p-JAK2/JAK2,p-STAT1/STAT1,p-STAT3/STAT3 were decreased,SOD,T-AOC were increased in Danshensu low-dose group,Danshensu high-dose group,positive control group(P<0.05).Compared with Danshensu low-dose group,R values were longer,Angle angles and MA values were decreased,CK-MB,cTnI,MDA,myocardial cell apoptosis index,p-JAK2/JAK2,p-STAT1/STAT1,p-STAT3/STAT3 were decreased,SOD and T-AOC were increased in Danshensu high-dose group and positive control group(P<0.05).Compared with positive control group,the R value of Danshenu high-dose group was prolonged,the Angle angles and MA value were decreased,the CK-MB,cTnI,MDA,myocardial apoptosis index,p-JAK2/JAK2,p-STAT1/STAT1,p-STAT3/STAT3 were decreased,and SOD and T-AOC were increased(P<0.05).There were no significant differences in the relative expression of JAK2,STAT1,and STAT3 mRNA in each group(P>0.05).Conclusion Danshinsu can improve TEG-related indicators and oxidative stress,with myocardial protective effect,which may play some regulatory role by inhibiting JAK/STAT signaling pathway.