Effects of caulophine on caffeine-induced cellular injury and calcium homeostasis in rat cardiomyocytes.
Caulophine is a novel fluorenone alkaloid isolated from the radix of Caulophyllum robustum Maxim. Caulophine showed high affinity for the rat myocardial cell membrane as assessed by cell membrane chromatography, suggesting that the compound may exert bioactivity in the heart. It is known that calcium plays an important role in the pathogenesis of ischaemic heart disease, and caffeine can cause calcium overload in cardiomyocytes by inducing calcium release from the sarcoplasmic reticulum. Therefore, the present study evaluated the effects of caulophine on caffeine-induced injury and calcium homeostasis in cardiomyocytes. Cardiomyocytes were pre-treated with caulophine before exposure to caffeine or potassium chloride (KCl). Cell viability was assayed using the MTT method, and lactate dehydrogenase (LDH) and malondialdehyde (MDA) were measured spectrophotometrically. Caulophine-pre-treated cardiomyocytes were incubated with Fluo-3/AM, and then caffeine or KCl was used to induce Ca(2+) overload. The total intracellular Ca(2+) concentration was measured by flow cytometry. Fluorescence densities of single cardiomyocytes were detected using a confocal microscope. Caulophine increased the viability of caffeine-injured cardiomyocytes and decreased LDH activity and MDA level in cardiomyocytes. Furthermore, caulophine significantly decreased the total intracellular free Ca(2+) concentration and intracellular calcium release in cardiomyocytes in response to caffeine. However, the same concentrations of caulophine did not affect KCl-induced calcium influx. Our results suggest that caulophine protects cardiomyocytes from caffeine-induced injury as a result of calcium antagonism. This finding provides a basis for further study and development of caulophine as a new calcium antagonist for treating ischaemic cardiovascular diseases.
Si
Kai-Wei
KW
Department of Pharmacology, Xi'an Jiaotong University School of Medicine, Xi'an, China.
Liu
Jun-Tian
JT
He
Lang-Chong
LC
Li
Xi-Kuan
XK
Gou
Wei
W
Liu
Chuan-Hao
CH
Li
Xiao-Qi
XQ
D000814:Aniline Compounds / Q000378:metabolism; D000818:Animals; D002110:Caffeine / Q000009:adverse effects*; D002118:Calcium / Q000378:metabolism*; D002470:Cell Survival; D002478:Cells, Cultured; D005449:Fluorenes / Q000494:pharmacology*; D005456:Fluorescent Dyes / Q000378:metabolism; D006706:Homeostasis; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000378:metabolism; D011189:Potassium Chloride / Q000378:metabolism; D051381:Rats; D012519:Sarcoplasmic Reticulum / Q000378:metabolism; D014966:Xanthenes / Q000378:metabolism
D000814:Aniline Compounds; D005449:Fluorenes; D005456:Fluorescent Dyes; D014966:Xanthenes; C554063:caulophine; C059715:Fluo-3; D002110:Caffeine; D011189:Potassium Chloride; D002118:Calcium