| Objective:Through investigating the influence of water extracting-alcohol precipitating solution of Shuganhewei decoction on gastrointestinal function in normal rats and gastrointestinal function, MTL, GAS and Gherlin in functional dyspepsia rats models with liver depression with spleen insufficiency, Screen out the best alcohol precipitation concentration. To study mechanism of extracting-alcohol precipitating solution of Shuganhewei decoction in treating functional dyspepsia with liver depression with spleen insufficiency through investigating its the influence on correlation hormone level and proteins, so as to provide experimental basis for development and clinical application of Shuganhewei oral liquid.Methods:To screen out the better alcohol precipitation concentration from different alcohol degree (30%,55%,80%) as indices of the rate of gastric residual and intestinal propulsion. Established rat model of functional dyspepsia with liver depression with spleen insufficiency by irregular diet and dilute hydrochloric acid fed and tail clamped. To screen out the best range of alcohol precipitation concentration through observing the influence of the better alcohol precipitation concentration of water extracting-alcohol precipitating solution of Shuganhewei decoction on gastrointestinal motility, MTL, GAS and Gherlin in model rats.Finally, studied on the effect of the best alcohol precipitation concentration on gastrointestinal function and the levels of serum SP, CCK,5-HT, ET-1, NO, Ghrelin, AChE, and to determine antrum Ghrelin and 5-HT protein by western blot and immunohistochemical method respectively.Results:1. The influence of water extracting-alcohol precipitating solution of Shuganhewei decoction (alcohol precipitation concentration:30%,55%,80%) on gastrointestinal motility in normal rats:compared with normal control group,55%alcohol precipitation control group and 80% alcohol precipitation control group all could obviously decrease gastric residual rate in normal rats (P<0.05).80% alcohol precipitation control group could significantly increase (P<0.05).2. The influence of the better alcohol precipitation concentration of water extracting-alcohol precipitating solution of Shuganhewei decoction on gastrointestinal motility, MTL, GAS and Ghrelin in functional dyspepsia rats models with liver depression with spleen insufficiency:(1) Weight:After the model was completed for two weeks, compared with normal control group (AG) rats, weight of model group (MG) rats had obviously decreased (P<0.01). After normal feeding, weight of model control group (BG) rats was still significantly lower than weight of AG rats (P<0.01). After one week of treatment, compared with BG rats, weight of 55% alcohol precipitation control group (DG) rats and 80% alcohol precipitation control group (EG) rats both increased significantly (P<0.01, P<0.05) except mosapride control group (CG) rats (P>0.05). After two weeks of treatment, weight of three treatment groups (CG, DG and EG) rats all higher than BG rats obviously (P<0.05, P<0.05,P<0.01). There was no statistical difference between the weight of AG, CG, DG and EG rats (P>0.05).(2) Gastrointestinal motility and the levels of serum MTL, GAS, Ghrelin:Compared with that in the AG, gastric residual rate in the BG had significantly increased (P<0.01), and intestinal propulsive rate, the levels of serum MTL, GAS, Ghrelin in the BG had significantly decreased (P<0.01). Compared with that in the BG, gastric residual rate in the DG and EG had obviously decreased (P<0.01), and intestinal propulsive rate (both P<0.01), the levels of serum MTL (P<0.01), GAS (P<0.01), Ghrelin (P<0.05,P<0.01) had significantly increased. Compared with that in the CG, gastric residual rate in the DG had significantly increased (P<0.01), and serum Ghrelin had obviously decreased (P<0.01), and there was no statistical difference between DG and CG in intestinal propulsive rate and the levels of serum MTL and GAS (P>0.05), and there was no statistical difference between EG and CG (P>0.05).3. Study on mechanism of 80% of alcohol precipitation concentration of water extracting-alcohol precipitating solution of Shuganhewei decoction in functional dyspepsia with liver depression with spleen insufficiency:(1) Weight:After the model was completed for two weeks,compared with normal control group (I G) rats, weight of model group (MG) rats had obviously decreased (P<0.01). After normal feeding, the weight of model control group (II G) rats was still significantly lower than the weight of I G rats (P<0.01). After one week of treatment, weight of mosapride control group (III G) rats and Shuganhewei control group (Ⅳ G) rats had obviously decreased compared with I G (P<0.01). After two weeks of treatment, weight of FVG rats was more than ⅡG (PO.01), and there no statistical difference between Ⅳ G and I G (P>0.05). Weight of ⅢG rats had decreased significantly compared with I G (P<0.05), and there was no statistical difference between IIIG and ⅡG (P>0.05).(2) Gastrointestinal motility, the levels of serum SP, CCK,5-HT, ET-1, NO, Ghrelin, AChE and the expressions of antrum Ghrelin and 5-HT:Compared with that in I G, gastric residual rate in the ⅡG had significantly increased (P<0.01), and intestinal propulsive rate (P<0.01), the levels of serum SP, CCK,5-HT, Ghrelin in the II G had significantly decreased (P<001, P<0.01,P<0.01,P<0.05), and the levels of NO and AChE had obviously increased (P<0.05), and the expressions of antrum Ghrelin showed a decreasing trend but no statistic difference (P>0.05), and the expressions of antrum 5-HT had significantly decreased. Compared with that in the Ⅱ G, gastric residual rate, the levels of serum NO, AChE in the IV G had obviously decreased (P<0.01, P<0.05,P<0.05), and intestinal propulsive rate, the levels of serum SP, CCK,5-HT, Ghrelin had significantly increased (P<0.05,P<0.01,P<0.01,P<0.05,P<005), and the expressions of antrum Ghrelin,5-HT were up--regulated (P<0.05). Compared with that in the IIIG, the levels of serum SP, CCK had significantly increased (P<0.01,P<0.05), and the level of serum Ghrelin had obviously decreased (P<0.05). The levels of serum ET-1 were no statistic difference between four groups (P>0.05). the expressions of antrum Ghrelin in the Ⅲ G was higher than that in the ⅣG. There was no statistic difference between ⅢG and ⅣG about the expression of autrum 5-HT (P>0.05).Conclusion:1.55% and 80% of alcohol precipitation concentration all can promote gastrointestinal motility in normal rats.2. It was used to make rat model of functional dyspepsia with liver depression with spleen insufficiency that irregular diet and dilute hydrochloric acid fed and tail clamped in two tests. The results show that this method is stable and feasible.3.55% and 80% of alcohol precipitation concentration all can promote gastrointestinal motility and enhance the levels of serum MTL, GAS, Ghrelin in model rats of functional dyspepsia with liver depression with spleen insufficiency, and the effect of 80% of alcohol precipitation concentration is better than the effect of 50% of alcohol precipitation concentration.4. Water extracting-alcohol precipitating solution of Shuganhewei decoction can promote gastrointestinal motility in model rats of functional dyspepsia with liver depression with spleen insufficiency. The possible mechanisms of the treatment of functional dyspepsia with liver depression with spleen insufficiency is that it regulates gastrointestinal smooth muscle movement, so as to enhance the role of gastrointestinal motility by upregulating the levels of related brain-gut peptides (MTL, SP, CCK,5-HT and Ghrelin), and then exciting cholinergic nerve, and regulates the balance of the digestive system by intrinsic nervous system and autonomic nervous system to impact on neurotransmitters (ACh, SP and NO) of motor neurons, and regulates stomach acid-base balance by regulating related hormones (ACh and GAS). |
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