The Effect Of Drug-Containing Serum Of Fructus Aurantii Immaturus On The Cell Contraction, Intracellular Calcium Concentration And Calmodulin Expression In Rat Antral Smooth Muscle

Objective.To observe the effect of drug-containing serum of Fructus Aurantii Immaturus (FAI) from SD rat on the effect of cell contraction, intracellular-free-calcium (Ca2+) concentration and calmodulin expression in antral smooth muscle.Methods:Rat drug-containing serum of FAI was prepared according to the common method of serum pharmacology. The serum was divided into 10%, 20%,50% drug-containing serum of FAI,20% domperidone group and no serum as a control group. The antral smooth muscle cells were extracted under dissecting microscope by enzyme collagenase and cultured at 37℃ in 5% CO2 incubator. The contraction effect of each group on antral smooth muscle cells was observed under inverted microscope. The changes of intracellular Ca2+ concentration in the antral smooth muscle before and after dosing in each group were conducted using confocal laser scanning microscopy. Immunohistochemical method was used to test the intracellular calmodulin expressions of antral smooth muscle in each group.Results:1. The effect of 10%,20% and 50% drug-containing serum of FAI on antral smooth muscle cell contraction and intracellular Ca2+fluorescence intensity showed a dose-dependent relationship. Cell contraction and intracellular Ca2+ fluorescence intensity increased with the elevation of drug concentration (P<0.05). When compared with blank serum control group,20% and 50% doses of FAI,20% domperidone group showed the significant difference of smooth muscle cell contraction and the intracellular Ca2+ concentration (P<0.05) but no difference in those of 10% dose group.There was no significantly different outcomes in the comparison of 10% doses of FAI versus blank serum control (P>0.05) as well as 20% FAI compared with 20% domperidone on cell contraction and Ca2+ concentration.2.10%,20% and 50% drug-containing serum of FAI showed the calmodulin expression rates 40%、53.3% and 86.7%,respectively. The positive rate depended on the increased concentration level. The antral smooth muscle intracellular calmodulin expressions presented a statistically significance between the 50%FAI and the blank serum control group (P<0.05). Neither 10%,20% of FAI drug-containing serum groups nor 20% domperidone group was no significant difference.Conclusion:The drug-containing serum of FAI can promote antral smooth muscle cell contraction. The mechanism may be related to the increase of intracellular Ca2+ concentration and the adjustment of calmodulin expression.