Zhang Jin-Yan, Li Peng, Li Yi-Kui
Experiment and Research Center, Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing 100091.
Zhongguo Zhong Xi Yi Jie He Za Zhi. 2012 Apr;32(4):510-4.
To observe the protective effects of paeonol, paeoniflorin, and their compatibility on in vitro cultured cardiomyocytes suffering from hypoxia-reoxygenation injury.
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.
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).
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.
观察丹皮酚、芍药苷及其配伍对体外培养的缺氧复氧损伤心肌细胞的保护作用。
体外培养新生大鼠心肌细胞，采用2.5 - 5小时缺氧及随后2小时复氧的方法造模。为观察丹皮酚和芍药苷的作用，分别设置100、75、50和25 mg/L四个剂量。为观察丹皮酚与芍药苷的配伍情况，设置五个剂量如下：丹皮酚40和20 mg/L、芍药苷40和20 mg/L、丹皮酚20 mg/L与芍药苷20 mg/L配伍。上述药物分别在缺氧及复氧期间与心肌细胞共同孵育。模型组不给予药物干预，正常对照组不造模。分别于缺氧期及复氧期结束后测定培养基中漏出的肌酸激酶（CK）、乳酸脱氢酶（LDH）和丙二醛（MDA），并计算整个过程中的总漏出量及漏出抑制率。每组结果均来自5次平行操作。
与正常对照组相比，缺氧2.5小时后MDA漏出增加，缺氧3、5小时及复氧2小时后CK、LDH和MDA的漏出量及总漏出量均显著增加。各指标的漏出抑制率降低，差异有统计学意义（P<0.01，P<0.05）。与模型组相比，高剂量丹皮酚组、高剂量和中剂量芍药苷组在缺氧后及复氧2小时后LDH和MDA的漏出量及总漏出量均降低（P<0.01，P<0.05），各指标的漏出抑制率升高（P<0.01，P<0.05）。然而，低剂量和极低剂量丹皮酚组仅在复氧2小时后CK的漏出量及总漏出量降低（P<0.01，P<0.05），而CK的漏出抑制率升高（P<0.01）。极低剂量芍药苷组仅在复氧2小时后LDH的漏出量及总漏出量降低（P<0.01），而LDH的漏出抑制率升高（P<0.01）。其配伍效果差异无统计学意义（P>0.05）。
丹皮酚、芍药苷及其配伍对体外培养的缺氧复氧损伤心肌细胞均有显著保护作用。丹皮酚与芍药苷联合使用时无明显协同作用。
