麦贞花颗粒对H_9C_2心肌细胞缺氧/复氧损伤的保护作用及凋亡的影响

Protective effect of serum containing Maizhenhua Granules on H9C2 cardiomyocytes exposed to hypoxia-reoxygenation

目的探讨麦贞花颗粒(麦冬、女贞子、灵芝、水蛭等)含药血清对H9C2心肌细胞缺氧/复氧损伤的保护作用及对细胞凋亡的影响。方法将H9C2心肌细胞随机分为正常对照组、缺氧/复氧组和麦贞花颗粒含药血清组。用酶联免疫吸附试验检测超氧化物歧化酶(SOD)、丙二醛(MDA)、环磷酸腺苷(cAMP),定磷法测定Na+-K+-ATP酶活性,RT-PCR测定谷胱甘肽过氧化物酶(GPX)mRNA表达,电镜检测心肌细胞凋亡。结果麦贞花颗粒血清预处理心肌细胞48 h能明显降低上清液中MDA水平,提高SOD、Na+-K+-ATP酶、cAMP活性,上调GPX mRNA表达,与缺氧/复氧组相比均有显著性差异(均P<0.05～0.01);电镜检测心肌细胞显示麦贞花颗粒含药血清组具有相对完整的细胞膜和细胞器而缺氧/复氧已经出现凋亡小体。结论麦贞花颗粒含药血清对心肌细胞缺氧/复氧损伤具有明显保护作用并能抑制心肌细胞凋亡。

AIM To explore the effect of serum containing Maizhenhua Granules （ Ophiopogonis Radix, Ligus- tri lucidi Fructus, Ganoderma, Hirudo, etc. ） on H9 C2 cardiomyocytes exposed to hypoxia-reoxygenation and cell apoptosis. METHODS H9C2 cardiomyocytes were divided into normal group, hypoxia-reoxygenation group and Maizhenhua Granules group. Enzyme linked immunosorbent assay was used to detect the levels of superoxide dis- mutase （SOD）, malondialdehyde （MDA） and cyclic adenosine monophosphate （cAMP）. Phosphorus measure- ment was used to detect the activity of Na ＋ -K ＋ -ATPase. RT-PCR was used to detect expression of glutathione per- oxidase （GPX） mRNA. Electron microscopy was used to detect cell apoptosis. RESULTS Maizhenhua Granules drug-serum pretreated after 48 h could obviously decrease the level of MDA, increase activity of SOD, Na ＋ -K ＋ ATPase, cAMP, and upregulate expression level of GPX mRNA as compared with those in hypoxia-reoxygenation group （ all P 〈 0. 05 - 0. 01 ）. Electron microscopy showed that cardiomyocytes in Maizhenhua Granules had rela- tive complete cell membrane and organelles, but had presented the apoptotic body in hypoxia-reoxygenation group. CONCLUSION Maizhenhua Granules have the protective effect on cardiomyocytes exposed to hypoxia-reoxygen- ation.