Role of bilobalide in ischemia reperfusion injury of skeletal muscle cells in rats / 中华创伤杂志

ABSTRACT

Objective:To examine the effects of bilobalide on ischemia reperfusion injury (IRI) of skeletal muscle cells in rats.Methods:IRI of skeletal muscle cells in L6 rats was stimulated by culturing skeletal muscle cells in a hypoxia incubator for 4 hours, which were further cultured in a conventional incubator for 4 hours. The induced rat myoblasts were randomly divided into control group, ischemia reperfusion group and bilobalide group. Rat myoblasts in control group were cultured in the conventional incubator, and cells in ischemia reperfusion group were cultured in hypoxia incubator and conventional incubator alternatively to simulate IRI of skeletal muscle cells. After 4 hours of incubation in hypoxic incubator, L6 rat myoblasts in bilobalide group were replaced with medium containing 20 μmol/L bilobalide, and then put into conventional incubator for further culture. CCK-8 assay was used to test cell viability 48 hours later. Related test kits were used to examine the levels of lactate dehydrogenase (LDH), superoxide dismutase (SOD), xanthine oxidase (XOD), malondialdehyde (MDA) and intracellular calcium.Results:Compared with ischemia reperfusion group [(62.2±6.8)%], the viability of myoblasts was increased in bilobalide group [(86.5±5.6)%] ( P<0.05). Compared with ischemia reperfusion group [(265.3±35.2)U/mg], the level of LDH in myoblasts was decreased in bilobalide group [(125.4±16.9)U/mg] ( P<0.05). Compared with ischemia reperfusion group [(68.6±10.7)U/mg], the level of SOD in myoblasts was increased in bilobalide group [(98.5±8.4)U/mg] ( P<0.05). Compared with ischemia reperfusion group [(224.7±65.8)U/mg], the level of XOD in myoblasts was decreased in bilobalide group [(105.9±58.9)U/mg] ( P<0.05). Compared with ischemia reperfusion group [(198.2±35.7)U/mg], the level of MDA in myoblasts was decreased in bilobalide group [(100.4±25.3)U/mg] ( P<0.05). Compared with ischemia reperfusion group (2.5±0.3), the intracellular Ca 2+ concentration in cell supernatant was decreased in bilobalide group (1.6±0.3) ( P<0.05). But all the indexes tested above did not return to the level in control group ( P<0.05). Conclusion:Bilobalide can protect skeletal muscle cells from IRI by inhibiting oxidative stress reaction and calcium overload.