Effect Of Banxia Xiexin Decoction On Cx43 - Mediated GJIC In Gastrointestinal Smooth Muscle Cells

The Banxiaxiexin Tang for purging stomach-fire is from Shang Han Za Bing Lun, and has downbear counterflow and check vomiting, dissolving lumps and resolving masses function. It is most used to epigastric oppression caused by misusing of Xiaochaihu Tang and vomiting sickness for a long time with borborygmus and epigastric oppression. The typhia master Mr Liu Duzhou described Fang Yi of the Banxiaxiexin Tang:" pinelliae and dried ginger warm pungently to eliminate cold of disposition; scutellaria baicalensis, coptis chinensis bitter cold vent to decrease heat of stomach qi; ginseng, liquorice and jujube sweet warm tone up to harmonize spleen and stomach and tonifying qi of middle-jiao energy to recover the function of middle burner upper abdomen. It is the pungent open and bitter down and sweet-nourishing method. " It has credible curative effect in treating gastrointestinal motor function disorder, which has been praised highly by many typhia masters and is one of the research focuses of Chinese medicine herbs.The article discussed the function mechanism of the Banxiaxiexin Tang to gastrointestinal smooth muscle cells Cx43 mediated GJIC to further explain molecular mechanism of its effect to the gastrointestinal motility and the separated prescriptions’ effects.We used serum pharmacology method to make serum containing the Banxiaxiexin Tang and the separated prescriptions by obtaining primary gastrointestinal smooth muscle cells of rats and steady culturing after identified by immunohistochemistry. Observe the effect of the Banxiaxiexin Tang and the separated prescriptions to cell proliferation of gastrointestinal smooth muscle using MTT method; observe the effect of the Banxiaxiexin Tang and the separated prescriptions to expression of gastrointestinal smooth muscle cells Cx43 mRNA,p38 MAPK mRNA, PKA mRNA and PKC mRNA using RT-PCR method; observe the effect of the Banxiaxiexin Tang and the separated prescriptions to expression of gastrointestinal smooth muscle cells Cx43. P-Cx43、p38 MAPK, P-p38MAPK、PKA and PKC using western blot method.MTT results:after 24h medicated serum effect, OD values of the decoction group, the pungent group, the bitter group, sweet-nourishing group and the pungent bitter group were higher than the blank group. Only sweet-nourishing group and the decoction pungent group had significance in statistics compared the blank group(P<0.05) The OD values of pungent sweet-nourishing group and the bitter sweet-nourishing group were lower than the decoction group and had significance in statistics (P<0.05).After 48h medicated serum effect, OD values of the decoction group, the pungent group, the bitter group, the bitter sweet-nourishing group were higher than the blank group and had significance in statistics compared the blank group(P＜0.01). The OD values of the pungent group, the bitter group and the bitter sweet-nourishing group were higher than the decoction group and the rest groups’OD values were lower than the decoction group and the pungent group and the pungent sweet-nourishing group had significance in statistics compared the decoction group(P<0.01), the pungent bitter group had significance in statistics compared the decoction group(P<0.05). After 72h medicated serum effect, OD values of every group were higher than the blank group and had significance in statistics(P<0.01).The OD value of the decoction group was lower than the pungent group, the bitter group, sweet-nourishing group and the pungent bitter group and the rest groups’OD values were lower than the decoction group and the pungent bitter group had significance in statistics compared the decoction group(P<0.05).RT-PCR results:there was no statistics significance between the blank group and the serum containing the Banxiaxiexin Tang and the separated prescriptions to the Cx43 mRNA gene expression. But there was effect trend. The gene expression enhancement of the pungent sweet-nourishing group was the strongest, followed by the sweet-nourishing group.The suppression expression of the pungent bitter group was the strongest, followed by the pungent group, the bitter group, the bitter sweet-nourishing group and the decoction group. The decoction group’s effects of actions were closed to the blank group, which was suppressive trend. There was no statistics significance to the p38 MAPK mRNA gene expression. But there was effect trend. The gene expression enhancement of the bitter sweet-nourishing group was the strongest, followed by the decoction group, the pungent sweet-nourishing group and the bitter group. The suppression expression of the pungent bitter group was the strongest, followed by the bitter sweet-nourishing group and the pungent group. The decoction group’s effect of actions was enhanced trend. There was no statistics significance to the PKA mRNA gene expression. But there was effect trend, which was suppressive trend for all groups. The gene expression suppression of the pungent bitter group was the strongest, followed by the sweet-nourishing group, the pungent group, the bitter sweet-nourishing group, the decoction, the pungent sweet-nourishing group and the bitter group. The decoction group’s effect of actions was suppressive trend. There was no statistics significance to the PKC mRNA gene expression. But there was effect trend, which was enhanced trend for all groups. The gene expression enhancement of the bitter sweet-nourishing group was strongest, followed by the bitter group, the pungent sweet-nourishing group, the pungent group, the decoction group, the pungent bitter group and the sweet-nourishing group. The decoction group’s effect of actions was enhanced trend.Western blot results:there was no statistics significance between the blank group and the serum containing the Banxiaxiexin Tang and the separated prescriptions to the Cx43 expression. But there was effect trend. The gene expression enhancement of the bitter sweet-nourishing group was the strongest, followed by the pungent sweet-nourishing group, the pungent bitter group, the decoction group and the sweet-nourishing group. The gene expression suppression of the bitter group was the strongest, followed by the pungent group. The decoction group’s effect of actions was enhanced trend. There was no statistics significance to the P-Cx43gene expression. But there was effect trend. The gene expression enhancement of the pungent bitter group was the strongest, followed by the bitter group, the sweet-nourishing group and the pungent sweet-nourishing group. The gene expression suppression of the bitter sweet-nourishing group was the strongest, followed by the pungent group and the decoction group. The decoction group’s effect of actions closed to the blank group, which was suppressive trend. There was no statistics significance to the p38 MAPK gene expression. But there was effect trend, which was suppressive trend for all groups. The gene expression suppression of the pungent group was the strongest, followed by the bitter group, the sweet-nourishing group, the bitter sweet-nourishing group, the pungent sweet-nourishing group, the pungent bitter group and the decoction. The decoction group’s effect of actions closed to the blank group, which was suppressive trend. There was no statistics significance to the P-p38 MAPK expression. But there was effect trend. The gene expression enhancement of the pungent bitter group was the strongest, followed by the sweet-nourishing group, the pungent group and the decoction group. The gene expression suppression of the bitter sweet-nourishing group was the strongest, followed by the pungent sweet-nourishing group and the bitter group. The decoction group’s effect of actions closed to the blank group, which was enhanced trend. There was no statistics significance to the PKA expression. But there was effect trend. The gene expression enhancement of the pungent bitter group was the strongest, followed by the pungent sweet-nourishing group and the bitter sweet-nourishing group. The gene expression suppression of the decoction group was the strongest, followed by the pungent group, the sweet-nourishing group and the bitter group. The decoction group’s effect of actions was suppressive trend. There was no statistics significance to the PKC expression. But there was effect trend. The gene expression enhancement of the bitter group was the strongest, followed by the decoction group. The gene expression suppression of the bitter sweet-nourishing group was the strongest, followed by the pungent bitter group, the pungent group, the sweet-nourishing group and the pungent sweet-nourishing group. The decoction group’s effect of actions was enhanced trend.The test results above showed that the Banxiaxiexin Tang and the separated prescriptions might play a role in regulating gastrointestinal smooth muscle contractions by enhancing or suppressing gastrointestinal smooth muscle cells Cx43、Cx43 mRNA and related protein phosphorylation expression and up or down regulating p38 MAPK,PKA,PKC signal transduction pathway.