丹皮酚、芍药苷及二者配伍对体外培养心肌细胞缺氧/复氧损伤的影响

Protective Effects of Paeonol,Paeoniflorin and Their Compatibility on in vitro Cultured Cardiomyocytes Suffering from Hypoxia-reoxygenation Injury

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中文关键词: 丹皮酚芍药苷心肌细胞缺氧/复氧

英文关键词:paeonol paeoniflorin cardiomyocyte hypoxia/reoxygenation

基金项目:国家自然科学基金青年科学基金项目(No.30801531)

作者	单位
张金艳	中国中医科学院西苑医院实验研究中心
李澎	中国中医科学院西苑医院实验研究中心
李贻奎	中国中医科学院西苑医院实验研究中心
赵乐	中国中医科学院西苑医院实验研究中心
贾广波	河北省三河市中医院急诊科

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中文摘要:

目的观察丹皮酚、芍药苷及二者配伍对体外培养心肌细胞缺氧/复氧损伤的保护作用。方法用体外细胞培养方法,培养乳鼠心肌细胞,复制心肌细胞缺氧/复氧损伤模型。分别在缺氧和复氧阶段将细胞培养液换成含药培养液以进行药物干预,研究丹皮酚和芍药苷的作用时,各设100、75、50和25mg/L4个剂量组,研究二者配伍作用时设芍药苷40mg/L和20mg/L组、丹皮酚40mg/L和20mg/L组、以及芍药苷20mg/L+丹皮酚20mg/L5个剂量组,模型组只造模不进行药物干预,而正常对照组则不造模。分别测定缺氧2.5～5h后及复氧2h后细胞培养上清液中的肌酸肌酶(creatine kinase,CK)、乳酸脱氢酶(lactate dehydrogenase,LDH)和丙二醛(malondialdehyde,MDA)漏出量,计算缺氧/复氧过程中各指标总漏出量及漏出抑制率。结果与正常对照组比较,模型组缺氧2.5h后MDA漏出量增加,缺氧3、5h后、复氧2h后CK、LDH和MDA漏出量、漏出总量均增加,各指标漏出抑制率均降低,差异均有统计学意义(P<0.01,P<0.05)。与模型组比较,丹皮酚高剂量组及芍药苷高、中剂量组缺氧后LDH、MDA漏出量及复氧2h后各指标漏出量、漏出总量均减少(P<0.01,P<0.05),各指标漏出抑制率均升高(P<0.01,P<0.05);而丹皮酚低、极低剂量组仅复氧2h后CK漏出量及漏出总量减少(P<0.01,P<0.05),CK漏出抑制率升高(P<0.01)。芍药苷极低剂量组仅复氧2h后LDH漏出量及漏出总量均减少(P<0.01),LDH漏出抑制率升高(P<0.01)。配伍组各指标与丹皮酚、芍药苷各组比较,差异无统计学意义(P>0.05)。结论丹皮酚、芍药苷及二者配伍对体外培养大鼠心肌细胞缺氧/复氧损伤均具有保护作用,二者配伍作用强度与同剂量单味药物作用强度相当,未见显著配伍增效作用。

英文摘要:

Objective To observe the protective effects of paeonol,paeoniflorin,and their compatibility on in vitro cultured cardiomyocytes suffering from hypoxia-reoxygenation injury.Methods Cardiomyocytes from neonatal rats were in vitro cultured and injured by a hypoxia of 2.5-5 h and a following 2-h reoxygenation.To observe the effects of paeonol and paeoniflorin,four doses of 100,75,50 and 25 mg/L were respectively set up.And to observe the compatibility of paeonol and paeoniflorin,five doses were set up as follows:paeonol 40 and 20 mg/L,paeoniflorin 40 and 20 mg/L,compatibility of paeonol 20 mg/L and paeoniflorin 20 mg/L.The above drugs were incubated with cardiomyocytes during the hypoxia and reoxygenation period respectively.No drug intervention was given to the model group,while no modeling was given to the normal control group.The transudatory creatine kinase(CK),lactate dehydrogenase(LDH),and malondialdehyde(MDA) in the culture medium were determined after the hypoxia period and the reoxygenation period respectively,and the total outleakage and the leakage inhibition ratio during the whole procedure were calculated.Results of each group were got from parallel operations for 5 times.Results Compared with the normal control group,the MDA leakage increased 2.5 h after hypoxia,the leakage and the total outleakage of CK,LDH,and MDA all significantly increased 3 and 5 h after hypoxia,and 2 h after reoxygenation.The leakage inhibition ratio of each index decreased with statistical difference(P<0.01,P<0.05).Compared with the model group,the leakage and the total outleakage of LDH and MDA both decreased in the high dose paeonol group,and the high and middle dose paeoniflorin groups after hypoxia and 2 h after reoxygenation(P<0.01,P<0.05),and the leakage inhibition ratio of each index increased(P<0.01,P<0.05).However,the leakage and the total outleakage of CK decreased in the low dose and the extreme low dose paeonol groups only 2 h after reoxygenation(P<0.01,P<0.05),while the leakage inhibition ratio of CK increased(P<0.01).The leakage and the total outleakage of LDH decreased in the extreme low dose paeoniflorin group only 2 h after reoxygenation(P<0.01),while the leakage inhibition ratio of LDH increased(P<0.01).The effects of their compatibility showed no significant difference(P>0.05).Conclusions Paeonol,paeoniflorin,and their compatibility all have remarkable protective effects on in vitro cultured cardiomyocytes suffering from hypoxia-reoxygenation injury.There was no significant synergistic effect when paeonol was used with paeoniflorin together.

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