Protection of total paeony glycoside on cardiomyocytic injury in neonatal rats cultured in vitro / 中国组织工程研究

ABSTRACT

BACKGROUND: It is demonstrated in modern pharmacologic study that total paeony glycoside (TPG) provides extensive pharmacologic activities,such as inhibiting aggregation of platelets and erythrocytes, anticoagulation,antithrombsis, anti-arterial sclerosis, protecting heart and liver, anti-tumor,etc.OBJECTIVE: Neonatal rat cardiomyocytes were cultured in vitro and by the changes of superoxide dismutase (SOD) activity, malondialdehyde (MDA) and nitric oxide (NO) contents in cell culture solution, the protection of TPG on injured cardiomyocytes was analyzed.DESIGN: Controlled observation was designed.SETTING: Bioengineering Department in School of Life Science and Technology of Xi 'an Jiaotong University and Institute of Bone Diseases in Medical School of Xi'an Jiaotong University.MATERIALS The experiment was performed in Bioengineering Department in School of Life Science and Technology of Xi 'an Jiaotong University from February to June in 2003, in which, 44 SD neonatal rats aged 1-3 days were employed. The 48-hour-cultured cardioryocytes were prepared in 42 bottles and randomized into 6 groups, named normal control (normal group), medicated-injury group (injury group), TPG 0.625 mg/L group,TPG 3.125 mg/L group, TPG 15.625 mg/L group and positive control, 7 bottles in each group.METHODS: Cardiomyocytic primary culture was performed under aseptic condition. No any drug was used in normal group, isoprenaline was added in injury group to terminate the concentration at 100 mg/L, in TPG 0.625, 3.125 and 15.625 mg/L groups, 30 minutes after isoprenaline added, RGP at dosages of 0.625, 3.125 and 15.625 mg/L were added respectively; in positive control, 30 minutes after isoprenaline added, coenzyme Q10 was used to terminate the concentration as 100 mg/L.Afterwards, the assay of every index was performed. Xanthine oxidase (XOD) method was used to assay SOD activity, thiobarbituric acid (TBA)method was to assay MDA content and nitrate reductase (NR) method was to assay NO content.cell culture solution in each group.Compared with normal group, the levels of total SOD, CuZn-SOD and MnSOD were reduced remarkably in injury group (P ＜ 0.05 or P ＜ 0.01).The above-indexes in every TPG group and positive control were improved to different extents (P ＜ 0.05 or ＜ 0.01), in which, the protection of TPG 15.625 mg/L group was near to or superior to positive control [(79.50±10.67), (80.30±13.50); (48.24±13.26), (49.73±10.23); (31.26±10.22),in cell culture solution in each group Those in injury group were higher remarkably than normal group (P ＜ 0.01). MDA and NO contents were all reduced in every TPG group and positive control and dose dependence presented in TPG protection, the higher the dose was, the stronger the action of TPG on protection was, in which, in high-dose group, MDA content was near to normal group [(5.41±1.81), (4.48±0.94) μmol/L, P ＞ 0.05] and NO content was similar to positive control [(81.83± 9.08), (82.41±12.37) mol/L,P ＞ 0.05].CONCLUSION: TPG protects myocardial injury induced by isoprenaline,indicating dose-dependence relationship, which is probably associated with enhanced anti-oxidation of cell, reduced injury of cellular membrane induced by free radical and lipid oxidant.