Effect Of Banxia Xiexin Decoction On CX43 - Mediated SMC And Its Inhibitory Transfection Of GJIC Function

ObjectiveFrom the view of cell biology to disscuss the significance of epigastric fullness and observe the molecular mechanism of gastrointestinal motility disturbance from Banxiaxiexin decoction.MethodsFirst obtained primary gastrointestinal smooth muscle cells by enzymatic digestion, and identified by immunohistochemical method and then cultured to the third generation for experiments. Second preparation containing serum of Banxiaxiexin decoction, Third, the experiment was divided into two parts. One part is to explore tne function of gap junction of the normal gastrointestinal smooth muscle cells, another part is to explore tne function of gap junction of the transfected by CX43 siRNA of normal gastrointestinal smooth muscle cells. So gastrointestinal smooth muscle cells CX43 siRNA model should be prepared for the follow-up to another part of the experiment. Fourth, the CCK-8 kit was used to observe the GJIC function between the normal gastrointestinal smooth muscle cells and the transfected by CX43 siRNA of normal gastrointestinal smooth muscle cells. Fifth, the apoptosis and Cell Cycle Kits were used to observe the situation of apoptosis and Cell Cycle between the normal gastrointestinal smooth muscle cells and the transfected by CX43 siRNA of normal gastrointestinal smooth muscle cells. Sixth, the use of RT-PCR was observed the situation of CX43mRNA, PKAmRNA, PKCmRNA, MAPKmRNA between the normal gastrointestinal smooth muscle cells and the transfected by CX43 si-RNA of normal gastrointestinal smooth muscle cells.ResultFirst,The results show that a simple Type ii collagenase digestion method can be successfully acquired gastrointestinal smooth muscle, the proposed enzymatic digestion was generally 35-45min, and this method could guarantee the amount of cell experiments. Second, By siRNA expression vector in gastrointestinal smooth muscle cells to stably inhibit the expression of CX43 model was successfully made, which providing an effective research tool for the study of CX43 and its GJ components in the gastrointestinal tract movement and its gastrointestinal tract diseases. Third, Banxiaxiexin decoction for normal SMC proliferation results:whole recipe group for SMC cells proliferation rate at 24h,48h,72h was 5.85%, 32.12% and 35.34%, showing a gradual upward trend. Forth, Banxiaxiexin decoction for SMC CX43 siRNA transfection proliferation results:At 24h, the whole recipe group compared for blank serum group was not statistically significant. At 48h, the whole recipe group compared for control group was statistically significant (P<0.05). At 72h, the whole recipe group compared for the blank group was not statistically significant. The overall trend of the whole recipe group and the control group was 72h OD value lower than 24h,48h. All recipe Group for SMC cells proliferation rate at 24h,48h,72h was respectively 10%, 32.5%,-2%. Fifth, Banxiaxiexin decoction, for normal SMC apoptosis results:early apoptotic rate of Banxiaxiexin decoction whole recipe group was higher than the control group, and there was significant difference (P<0.05), the apoptosis rate of the whole recipe group was higher than the control group, and there was a significant difference (P<0.05). Sixth, Banxiaxiexin decoction for SMC CX43 siRNA transfection apoptosis results:early apoptotic rate of the whole recipe group was lower than the blank serum group and there was significantly difference (P<0.05); the apoptosis of the whole recipe group lower than the blank serum group and there was compared statistically significance. Seventh, Banxiaxiexin decoction for cycle of SMC can be drawn, mainly cells were arrested in Gl phase, the whole recipe group was lower than the control group S phase, and there was statistically significance.Eighth, Banxiaxiexin decoction for CX43 suppressed SMC cycle experiments can be concluded that cells were arrested in Gl phase, whole recipe group in the G2 period was higher than the blank serum group, and there was statistically significance; the transfected whole recipe group in G2 phase arrest rate was half higher than normal non-transfected SMC. Ninth, in SMC, Banxiaxiexin decoction effected gap junction function of cells by inhibiting the expression of PKAmRNA, stimulating PKCmRNA, MAPKmRNA. After inhibited CX43, Banxiaxiexin decoction effected gap junction function of cells by stimulating the expression of PKAmRNA. PKCmRNA. MAPKmRNA.ConclusionBanxiaxiexin decoction can promote the express of PKAmRNA、PKCmRNA MAPKmRNA,so it can adjust SMC CX43 siRNA gaps junctions between gastrointestinal smooth muscle cell function. Banxiaxiexin decoction can depress of PKAmRNA, promote the express of PKCmRNA、MAPKmRNA,so it can adjust SMC gaps junctions between gastrointestinal smooth muscle cell function.