Objective To explore the possible mechanism of Liujunzi Decoction（六君子汤） plus Xuanfu Daizhe Decoction（旋覆代赭汤） in the treatment of gastroesophageal reflux disease. Methods Wistar rats aged 2 months were divided into low

medium and high dose of Chinese medicine groups

a mosapride group and a blank group

with10 rats in each group. The low

medium and high doses of Chinese medicine were administered with the concentration of 0. 846

1. 728

3. 456 g/ml Liujunzi Decoction（六君子汤） plus Xuanfu Daizhe Decoction（旋覆代赭汤）. The mosapride group was given a concentration of 0. 5 mg/ml mosapride solution

1. 5 ml/100 g per time

twice a day

and the blank group was given the same amount of normal saline for 5 days. On the 5 th day

1 hour after intragastric administration

blood was taken from the abdominal aorta to prepare drug-containing serum. Three Wistar rats born for 10 to15 days were fasting for 18 hours

about 2 cm gastric sinus tissue was taken out to culture the gastric interstitial cells（ICC） of rat

and identified by immunofluorescence. After 7 days of stable culture

ICC cells were divided into low

medium and high dose groups of Chinese medicine

mosapride group and blank group

with 12 samples in each group. The concentration was adjusted to 5 × 10

4

/ml and inoculated into 96-well plates. After 12 hours

the corresponding drug-containing serum was added to 200 μl/well for 24

48 and 72 h. The activity of ICC cells was detected by MTT assay. Fluo-3-AM fluorescence was used to detect changes in intracellular Ca

2+

concentration under laser confocal microscope. Results After 7 days of stable culture

the ICC showed a triangular

fusiform or polygonal shape under the inverted microscope. The nucleus was large and surrounded by cytoplasm. The cell body emitted a number of dendrites of various sizes

and the protrusions were connected with the cell bodies and protrusions of the surrounding cells. After immunofluorescence staining

the C-kit surface of ICC was positive under the microscope

and the cell bodies and protuberances were obviously stained

showing a positive green color. Compared with the blank group and the low-dose group of Chinese medicine

the ICC activity of the middle-and high-dose groups and the mosapride group were significantly increased at 24

48

and 72 h

and the Ca

2+

fluorescence intensity was increased（P

<

0. 05）. The ICC activity of the mosapride group was significantly higher than that of the middle and high dose groups of Chinese medicine at 48 h and 72 h. The fluorescence intensity of Ca

2+

was also higher than that of the middle and high dose groups（P

<

0. 05）. Conclusion Liujunzi Decoction（六君子汤） and Xuanfu Daizhe Decoction（旋覆代赭汤） can enhance the concentration of Ca

2+

in ICC and increase the activity of ICC

which may be one of the possible mechanisms for the treatment of gastroesophageal reflux disease.