摘要
目的:分析丹酚酸通过AKT/mTOR/p70S6K信号通路对骨骼肌缺血再灌注损伤小鼠细胞自噬及凋亡的影响。方法:选取SPF级雄性大鼠45只,随机数字表法分为假手术组、模型组、丹酚酸组,模型组、丹酚酸组制备骨骼肌缺血再灌注损伤模型,丹酚酸组分别在开始实验前72 h、48h、24 h及再灌注前15 min,腹腔注射剂量为30mg/kg的丹酚酸溶液1 mL,假手术组和模型组大鼠腹腔注射生理盐水1 mL。观察各组大鼠腓肠肌组织病理形态,血清肌酸激酶(CK)、乳酸脱氢酶(LDH)含量,腓肠肌组织Akt、p-Akt、p70S6K、p-p70S6K、mTOR及p-mTOR蛋白表达及腓肠肌组织内Bcl-2、Bax表达。结果:再灌注0 h、4 h及24 h时模型组大鼠腓肠肌湿/干重(W/D)比值较假手术组升高,丹酚酸组腓肠肌W/D比值较模型组降低,差异有统计学意义(P<0.05);再灌注0 h、4 h及24 h时模型组大鼠血清LDH、CK含量较假手术组升高,丹酚酸组大鼠血清LDH、CK含量较模型组降低,差异有统计学意义(P<0.05);模型组大鼠腓肠肌Akt蛋白表达较假手术组降低,p-Akt蛋白表达较假手术组升高;丹酚酸组大鼠Akt、p-Akt、p70S6K、p-p70S6K、mTOR及pmTOR蛋白表达较模型组升高,差异有统计学意义(P<0.05);光密度值(IOD)量化腓肠肌细胞内Bax、Bcl-2蛋白表达,模型组Bcl-2 IOD数值、Bcl-2/Bax比值较假手术组降低,Bax IOD数值较假手术组增加;丹酚酸组Bcl-2 IOD数值、Bcl-2/Bax比值较模型组升高,Bax IOD数值较模型组降低,差异有统计学意义(P<0.05)。结论:丹酚酸可维持骨骼肌缺血再灌注损伤大鼠骨骼细胞Bax与Bcl-2动态平衡,抑制细胞凋亡,其作用机制可能和激活AKT/mTOR/p70S6K信号通路有关。
AIM:To analyze the effects of salvianolic acid on autophagy and apoptosis of skeletal muscle ischemia-reperfusion injury mice through AKT/mTOR/p70S6K signaling pathway.METHODS:A total of 45 SPF male rats were randomly divided into sham operation group,model group,salvianolic acid group.Model group and salvianolic acid group were prepared for skeletal muscle ischemia-reperfusion injury model.At 72 h,48 h,24 h and 15 min before reperfusion,1 mL of salvianolic acid solution was administered intraperitoneally at a dose of 30 mg/kg.Rats in the sham operation group and the model group were intraperitoneally injected with 1 mL of normal saline.The pathological morphology of the gastrocnemius muscle,the contents of serum CK and LDH,the expression of Akt,p-Akt,p70S6K,p-p70S6K,mTOR and p-mTOR in the gastrocnemius muscle and the expression of Bcl-2 and Bax in the gastrocnemius muscle were observed.RESULTS:At the 0 h,4 h and 24 h within reperfusion,the W/D ratio of gastrocnemius muscle in the model group were higher than those in the sham operation group.The W/D ratio of the gastrocnemius muscle in the salvianolic acid group was lower than that in the model group(P<0.05).The levels of serumLDH and CK in the model group were higher than those in the sham operation group at 0 h,4 h and 24 h after reperfusion.The serum levels of LDH and CK in the salvianolic acid group were lower than those in the model group(P<0.05).The expression of Akt protein in the gastrocnemius of the model group was lower than that in the sham operation group,and the expression of p-Akt protein was higher than that in the sham operation group.The protein expressions of Akt,p-Akt,p70S6K,pp70S6K,mTOR and p-mTOR in the salvianolic acid group were significantly higher than those of the model group(P<0.05).The IOD value quantified the expression of Bax and Bcl-2 protein in the gastrocnemius muscle cells.The Bcl-2 IOD value and Bcl-2/Bax ratio in the model group were lower than those in the sham operation,whlie the Bax IOD value in the model group was higher t
作者
沈芸
张占丰
王红灵
赵冀安
SHEN Yun;ZHANG Zhanfeng;WANG Hongling;ZHAO Ji'an(The First People's Hospital of Huzhou,the First Affiliated Hospital of Huzhou Teachers College,Huzhou 313000,Zhejiang,China;Clinical Laboratory of Shandong Normal University,Jinan 250013,Shandong,China;Department of Hepatobiliary Surgery,the First Hospital of Hebei Medical University,Shijiazhuang 050050,Hebei,China)
出处
《中国临床药理学与治疗学》
CAS
CSCD
2020年第10期1111-1118,共8页
Chinese Journal of Clinical Pharmacology and Therapeutics
基金
河北省卫生和计划生育委员会课题项目(20150634)。
