| Objective: This study is to explore the effect of Citrus aurantium on the SCF/c-Kit signaling pathway and the proliferation of Interstitial Cells of Cajal(ICCs)in the antrum of rats with functional dyspepsia(FD).Methods:(1)Totally 48 healthy SD rats were randomly divided into 6groups: the normal group,the model group,the domperidone group(0.30 mg.ml-1),and the low/middle/high dose group of Citrus aurantium(0.5g.ml-1?1.0g.ml-1and 2.0g.ml-1 separately),8 in each group.(2)Except for the normal group,other five groups were devised model of FD rats which were stimulated with improved tail clamp method.The model of FD rats were treated with domperidone suspension(0.30 mg.ml-1)and Citrus aurantium decoction of low,middle,high dose(0.50 g.ml-1,1.0g.ml-1and 2.0g.ml-1 concentrations)to carry out intragastric administration respectively to the four groups for 4 weeks.(3)After 4 weeks,gastric tissue of all groups were taken for observing.Thehistological changes of gastric tissue were evaluated by HE staining.Gastric emptying of rats were recorded and the contents of SCF in serum were detected by ELISA.The transmission electron microscope was for observing the ultrastructure of ICCs,and the expression of c-Kit and SCF in gastric antrum of rats were observed by immunofluorescence.The expression of c-Kit mRNA and SCF mRNA in gastric antrum of rats were detected using RT-PCR respectively.The c-Kit protein and SCF protein expression in gastric antrum of rats were detected by Western blot.Results:(1)The model group had much gastric contents,the normal group,the low/middle/high dose group of Citrus aurantium and the domperidone group had less gastric contents.(2)Gastric tissue of all groups had no significant flushing,swelling,erosion,ulcer and other diseases.(3)The comparison of the gastric emptying rates of rats of all the groups: Compared with the normal group,the gastric emptying rate decreased in model group(p<0.05).Compared with the model group,the gastric emptying rates of Citrus aurantium groups(0.50 g.ml-1,1.0g.ml-1and 2.0g.ml-1 concentrations)and the domperidone group all improved(all p<0.05).Compared with the Citrus aurantium low-dose group,the gastric emptying rates in the middle and high dose groups of Citrus aurantium and the domperidone group all increased(all p<0.05).Compared with the Citrus aurantium middle-dose group,the gastric emptying rates all increased in the high dose group of Citrus aurantium and the domperidone group(all p<0.05).(4)The ultrastructure of ICCs of all the groups: The normal group had more ICCs,bigger size.In the model group,ICCs declined in the number and had smaller size with organelle degeneration,mitochondrial swelling or even vacuolation,and with surrounding cells and nerve fibers of peripheral network disordered and broken.The ultrastructure of ICCs improveddifferently in the low/middle/high dose group of Citrus aurantium and the domperidone group.(5)The comparison of contents of SCF in serum of rats of all the groups: Compared with the normal group,the SCF level in serum decreased dramatically in model group(p<0.01).Compared with the model group,the SCF level in serum in the Citrus aurantium low-dose group increased(p<0.05),and the SCF level in serum elevated significantly in the middle/high dose group of Citrus aurantium and the domperidone group(all p<0.01).Compared with the domperidone group,the SCF level in serum increased in the Citrus aurantium high-dose group(p<0.05).Compared with the Citrus aurantium low-dose group,the SCF level in serum in the Citrus aurantium middle-dose group and the domperidone group all increased(all p<0.05),the SCF level in serum elevated significantly in the high dose group of Citrus aurantium(p<0.01).Compared with the Citrus aurantium middle-dose group,the SCF level in serum in the Citrus aurantium high-dose group increased(p<0.05).(6)The expression of c-Kit and SCF in gastric antrum of rats were observed by immunofluorescence: Compared with the normal group,the mean fluorescence intensity of c-Kit and SCF in gastric antrum both decreased obviously in the model group(both p<0.01).Compared with the model group,the mean fluorescence intensity of c-Kit and SCF in gastric antrum both increased in the Citrus aurantium low-dose group(both p<0.05),and the mean fluorescence intensity of c-Kit and SCF in gastric antrum all elevated significantly in the middle/high dose group of Citrus aurantium and the domperidone group(all p<0.01).Compared with the domperidone group,the mean fluorescence intensity of SCF in gastric antrum increased in the Citrus aurantium high-dose group(p<0.05).Compared with the Citrus aurantium low-dose group,the mean fluorescence intensity of c-Kit and SCF in gastricantrum all increased in the Citrus aurantium middle-dose group and the domperidone group(all p<0.05),and the mean fluorescence intensity of c-Kit and SCF in gastric antrum both elevated significantly in the Citrus aurantium high-dose group(both p<0.01).Compared with the Citrus aurantium middle-dose group,the mean fluorescence intensity of SCF in gastric antrum increased in the Citrus aurantium high-dose group(p<0.05).(7)The expression of c-Kit mRNA and SCF mRNA in gastric antrum of rats were detected using RT-PCR respectively: Compared with the normal group,the mRNA expression of c-Kit and SCF in gastric antrum both decreased dramatically in the model group(both p<0.01).Compared with the model group,the mRNA expression of c-Kit and SCF in the Citrus aurantium low-dose group and the mRNA expression of SCF in the domperidone group all increased(all p<0.05),and the mRNA expression of c-Kit and SCF in the middle/high dose group of Citrus aurantium and the mRNA expression of c-Kit in the domperidone group all increased significantly(all p<0.01).Compared with the domperidone group,the mRNA expression of SCF increased in the Citrus aurantium middle-dose group(p<0.05).The mRNA expression of c-Kit and SCF both increased significantly in the Citrus aurantium high-dose group(both p<0.01).Compared with the Citrus aurantium low-dose group,the mRNA expression of c-Kit and SCF in the Citrus aurantium middle-dose group and the mRNA expression of c-Kit in the domperidone group all increased(all p < 0.05),and the mRNA expression of c-Kit and SCF both increased significantly in the Citrus aurantium high-dose group(both p < 0.01).Compared with the Citrus aurantium middle-dose group,the mRNA expression of SCF increased in the Citrus aurantium high-dose group(p<0.05),and the mRNA expression of c-Kit increased significantly in the Citrus aurantium high-dose group(p<0.01).(8)The c-Kit protein and SCF protein expression in gastric antrum of rats were detected by Western blot: Compared with the normal group,the c-Kit protein and SCF protein expression in gastric antrum both decreased dramatically in the model group(both p<0.01).Compared with the model group,the c-Kit protein and SCF protein expression in gastric antrum both increased in the Citrus aurantium low-dose group(both p<0.05),and the c-Kit protein and SCF protein expression in gastric antrum all increased significantly in the middle/high dose group of Citrus aurantium and the domperidone group(all p<0.01).Compared with the domperidone group,the c-Kit protein in the middle/high dose group of Citrus aurantium and the SCF protein expression in the Citrus aurantium high-dose group all increased(p<0.05).Compared with the Citrus aurantium low-dose group,the c-Kit protein in the domperidone group and the SCF protein in the Citrus aurantium middle-dose group and the domperidone group all increased(all p<0.05).The c-Kit protein in the Citrus aurantium middle-dose group and c-Kit protein and SCF protein in the Citrus aurantium high-dose group all increased significantly(all p<0.01).Compared with the Citrus aurantium middle-dose group,the SCF protein in the Citrus aurantium high-dose group increased(p<0.05).Conclusions: Citrus aurantium can improve gastric motility of rats with functional dyspepsia.It promoted gastric motility might be connected with the mechanisms of increasing the expression of c-Kit and SCF,starting SCF/c-Kit signaling pathways,promoting proliferation of ICCs,and improving the ultrastructure of ICCs. |
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