Experimental Study Of The Mechanism:Traditional Chinese Medicine Weichangshu Film To Promote Gastrointestinal Motility

[Objective] Through observe the effect of weichangshu on gastric emptying、gastric emptying to gastrointestinal motility disorder mice caused by Atropine、the effect of weichangshu to gastrointestinal hormone secretion about gastrointestinal motility disorder SD rat caused by L-Arg and its influence on the release of NO、the expression of eNOSmRNA in gastrointestinal smooth muscle cells of healthy SD rats.From different levels and in-depth exploration of progressively the role of Weichangshu film to promote the mechanism of gastrointestinal motility.[Methods] Choose kunming mice and were randomly divided into blank control group, model group atropine, weichangshu+atropine group, cisapride+atropine group, muxiangshunqi pills+atropine group. Control group and atropine model group are given the saline,the other groups were treated with oral liquid.For5d, after18h fasting, the sixth day in blank control group by intraperitoneal injection of saline.Other groups were injected with atropine sulfate injection to make animal model of gastrointestinal motility disorder.blue dextran (BD)2000for the marker to observe the gastric emptying rate and rate of intestinal propulsion.Select SD rats were randomly divided into normal control group、L-Arg group、L-Arg plus Weichangshu group and L-Arg plus xishabili group. In addition to the normal control group, the other three groups Intraperitoneal injection of L-Arg, preparation of animal models of gastrointestinal motility disorders. Normal control group was given normal saline, the other groups were given equal volume of drug administered, Abdominal aortic blood,Radioimmunoassay determination of plasma MTL, SP, SS content.Primary culture gastrointestinal smooth muscle cells with tissues, after successful identification, choose logarithmic growth phase of the third generation of cells, Experiment was divided into Weichangshu low-dose group、Weichangshu middle-dose group、Weichangshu high-dose group、 Xishabili group and normal control group. Interfere with the normal control group equivalent medium, tested drugs were used in the other groups interfere with cell24h, reaction with DAF-FM DA30min, detect the release of NO use Flow cytometry. extraction of total RNA after groups with different subjects of drug interference, use reverse transcription-polymerase chain reaction, semi-quantitative analysis the changes of the expression of eNOS mRNA levels in gastrointestinal smooth muscle cells, and conduct comparative analysis between groups.[Results]The gastric emptying rate and intestinal promoting rate of Atropine model group were significantly lower than the control group (P<0.01), Weichangshu and cisapride can make gastric emptying and intestinal propulsion have significant role (P<0.01) on gastrointestinal motility disorder animal caused by atropine.Model group MTL, SP, SS were significantly lower, compared with the normal group was statistically significant(P<0.05), after treatment,Weichangshu group MTL, SP, SS in plasma increased significantly, compared with model group was significant(P<0.01)。Compared with the normal control group, fluorescence intensity of NO release Significantly reduced in Gastrointestinal smooth muscle cells in Weichangshu high-dose group and the middle-dose group(P<0.01, P<0.05), eNOS mRNA expression was significantly lower in Weichangshu high-dose group and the middle-dose group(P<0.01, P<0.05).[Conclusion] The mechanism of Weichangshu promoting gastrointestinal motility:①Through the confrontation of atropine on the M receptor antagonism, increase gastrointestinal excitatory neurotransmitter acetylcholine (Ach) secretion, acetylcholine receptor on the M, the activation phospholipase C (PLC) β-coupled G protein, phospholipid hydrolysis Inositol bisphosphate to produce inositol triphosphate (IP3) and diacylglycerol (DAG), IP3as a second messenger intracellular calcium can make the Ca2+concentration, Ca2+signaling system triggered a series of reactions, so that gastrointestinal smooth muscle Contraction, thereby promoting gastrointestinal motility.(2)Through regulating the coordinated balance of gastrointestinal hormones in Plasma so as to promote gastrointestinal motility.(3)By increasing plasma MTL and SP content, cause the concentration of intracellular Ca2+in Gastrointestinal smooth muscle increase, Ca2+signaling system trigger a series of physiological functions and then promot the effect of gastrointestinal smooth muscle contraction.④The mechanism of Weichangshu gastrointestinal motility-promoting is may be inhibit the release of NO in gastrointestinal smooth muscle cells, and then inhibiting the activation of enzymes, antagonized the formation of cGMP, Thus increasing intracellular Ca2+, Ca2+signaling system triggered a series of physiological functions, Thereby promoting effects of gastrointestinal smooth muscle contraction.