龙血竭对体外心肌缺血再灌注树鼩模型细胞凋亡及内质网应激蛋白表达的影响  

Effects of Resina Dracoins in the proteins related to cell apoptosis and endoplasmic reticulum stress in tree shrews’ myocardial ischemia reperfusion modeling in vitro

作　　者：杨天睿[1] 张海燕[2] 叶堃 杨晶晶[1] 陈宇[1] YANG Tian-rui;ZHANG Hai-yan;YE Kun;YANG Jing-jing;CHEN Yu(The First People’s Hospital of Yunnan Province,Kunming 650032,China)

机构地区：[1]云南省第一人民医院,昆明理工大学附属医院,昆明650032 [2]云南省第一人民医院,昆明理工大学附属医院老年医学科,昆明650032

出　　处：《中华中医药杂志》2021年第12期7037-7042,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：云南省卫生科技计划项目(No.2016NS203)。

摘　　要：目的:研究龙血竭对体外心肌缺血再灌注树鼩模型B淋巴细胞瘤-2蛋白(Bcl-2)、Bcl-2相关X蛋白(Bax)、葡萄糖调节蛋白78/免疫球蛋白结合蛋白(GRP78)、内质网应激相关分子CCAAT/增强子结合蛋白同源蛋白(CHOP-1)、磷酸化P38(p-P38)、磷酸化应激活化蛋白激酶-1(p-JNK-1)、磷酸化细胞外调节蛋白激酶(p-ERK)蛋白表达量的影响。方法:采用Langendorff离体心脏灌流系统建立实验树鼩心肌缺血后龙血竭再灌注模型,将60只树鼩随机分为空白对照组,模型对照组,溶媒对照组和龙血竭低、中、高剂量组(0.25、0.5、1.0g/L),共6组,每组实验成功5只;空白对照组持续灌注60min,其余5组均停灌30min,缺血后再灌注30min,空白对照组和模型对照组的灌注液为K-H溶液,溶媒对照组的再灌注液为K-H溶液+聚山梨酯-80+95%乙醇,龙血竭低、中、高剂量组的再灌注液分别为0.25、0.5、1.0g/L的龙血竭溶液+K-H溶液。灌注结束后,Western Blot测定Bcl-2、Bax、GRP78、CHOP-1、p-P38、p-JNK-1、p-ERK蛋白表达量。结果:与模型对照组和溶媒对照组比较,龙血竭各剂量组Bcl-2表达量显著升高、Bax表达量显著降低(P<0.01),p-P38、p-JNK-1和p-ERK蛋白表达水平均显著降低(P<0.05,P<0.01),GRP78和CHOP-1蛋白表达水平均显著降低(P<0.01)。结论:龙血竭灌注通过上调BCL-2蛋白表达,下调Bax、GRP78、CHOP-1、p-P38、p-JNK-1、p-ERK蛋白表达,减少细胞凋亡,抑制内质网应激,进而达到抗心肌缺血再灌注损伤作用。Objective: To investigate the effects of Resina Dracoins on expression of BCL-2, Bax, GRP78, CHOP-1,p-P38, p-JNK-1 and p-ERK in tree shrews’ myocardial ischemia reperfusion modeling in vitro. Methods: The experimental tree shrews’ myocardial ischemia reperfusion model was established by Langendorff heart perfusion in vitro system. Sixty tree shrews were randomly divided into 6 groups, namely blank group, model group, solvent control group and low-dose, medium-dose, highdose Resina Dracoins(0.25, 0.5, 1.0 g/L) group, there were 5 successful tree shrews’ models in each group. Blank group keep continuous perfusion for 60 minutes, while other groups stop perfusion for 30 minutes and then start reperfusion for 30 minutes.The perfusate of blank group and model group is K-H solution and perfusate of solvent control group is mixed with Tween 80 and 75% alcohol;the perfusate of drug groups are different concentration(0.25, 0.5, 1.0 g/L) Resina Dracoins dissolved in Tween 80 and alcohol. After perfusion, expression of Bcl-2, Bax, GRP78, CHOP-1, p-P38, p-JNK-1 and p-ERK are detected by Western Blot. Results: Compared with the model control group and the vehicle control group, the expression of Bcl-2 in each dosage group of Resina Draconis significantly increased, while the expression of Bax significantly decrease(P<0.01), and the expression levels of p-P38, p-JNK-1 and p-ERK proteins in each dosage group of Resina Draconis significantly decreased(P<0.05, P<0.01), and the expression levels of GRP78 and CHOP-1 proteins were significantly decreased(P<0.01). Conclusion: Resina Dracoins has an effect against to myocardial ischemia-reperfusion injury by down-regulated expression of BCL-2 and up-regulated expression of Bax, GRP78, CHOP-1, p-P38, p-JNK-1 and p-ERK.

关 键 词：龙血竭 抗心肌缺血再灌注损伤 分子机制 

分 类 号：R285.5[医药卫生—中药学]