| Objective: Diabetes gastroparesis(DGP) is a kind of secondary to diabetes,on the basis of evil Heart, ictal vomiting, anorexia, early satiation and postprandial abdominal distension as the main clinical manifestations; Under the condition of the exclusion of mechanical obstruction of stomach gastric dilatation, slow gastric motility and gastric emptying delay is a common complication in diabetic patients, patients with serious damage to health. It is reported that diabetes incidence of DGP as much as 30%- 50%. In recent years, many reports on the pathogenesis of diabetic gastroparesis, have different effects, primarily with neuropathy, hyperglycemia, gastrointestinal hormone disorder, Cajal interstitial cells(ICC) lesions and microvascular lesion, gastrointestinal smooth muscle disease and abnormal factors such as the signal path.In recent years, the study found that abnormal gastrointestinal smooth muscle lesions and signaling pathways in the development of diabetic gastroparesis play a very important role. Gastrointestinal smooth muscle is the core of gastrointestinal power units, various factors ultimately through the gastrointestinal tract smooth muscle to regulate gastrointestinal dynamic role into full play. Diabetic gastroparesis in rat gastric electrical and mechanical coupling loss phenomenon is prompt DGP rats gastric antrum smooth muscle itself may be abnormal. Rho A/ROCK signaling pathway is a Ca2 +adjustment mechanism of the dependence of smooth muscle contraction, the effect of the main molecular include myosin light chain phosphatase(MLCP),myosin light streptokinase(MLCK) and phosphorylation of myosin light chain(p-MLC), such as the signal path can be adjusted by smooth muscle contraction after activation. Now we find it with vascular smooth muscle, bladder smooth muscle, there is a close relationship between abnormal contraction of uterine smooth muscle, etc, It is reported that the over expression of Rho A/ROCK may aggravate diabetic cardiomyopathy, causing abnormal vascular smooth muscle contraction, can participate in endothelial cell contraction, increase endothelial cell permeabilitybut, whether gastric smooth muscle may adjust the Rho A/ROCK signaling pathway is involved in the pathogenesis of DGP is unclear. This experiment was to observe Chinese traditional medicine for activating blood circulation to improve diabetes gastroparesis whether rat gastric dynamics associated with gastric smooth muscle of Rho A/ROCK signaling pathways, explore the possible mechanism of its therapeutic DGP for the clinical treatment of diabetes, mild stomach paralysis provide a theoretical basis.Methods: Sprague Dawley SD 30 rats, body weight of 201- 205 grams, adaptive feeding for 2 weeks were randomly divided into normal control group, the model group, and the traditional Chinese medicine group(leech, earthworm, salvia miltiorrhiza, rhizoma ligustici wallichii, etc.), Normal group and model group were given intraperitoneal injection of streptozotocin(STZ)25mg/kg, before using STZ, According to the proportion of normal 1% was diluted with citrate buffer(p H4,0.1 mmol/L). 72 h and 1wk after the injection, blood was collected at the tail vein of rats,Only random blood glucose is not less than 16.7mmol/L can determine the animal model was successful. After the success of the building, the normal group and model group were given saline through stomach feeding, Chinese traditional medicine group was given Chinese traditional medicine to fill the stomach, bought in Hebei province hospital of TCM pharmacy, Chinese medicine formulations for bags from decoction, and according to the weight of rats, were given traditional Chinese medicine(TCM) 10ml/(kg·d);once a day for 6 weeks. In the sixth week, the rats were treated by intragastric administration with phenol red solution(500mg/L) 2ml. After the anesthesia by intraperitoneal injection of 8% 2ml/kg amobarbital sodium, opened the abdominal cavity, ligated cardia and pylorus, removed the rat stomach and measured gastric emptying rate(GER) by phenol red. Take part in stomach tissue, placed in 4% paraformaldehyde, pathological changes of rats in each group were used to observe the gastric HE staining;Take part in gastric tissue quick-frozen in liquid nitrogen in the low temperature of-80 DEG C refrigerators, by RT-PCR method to detect gene expression in gastric tissue of Rho A, ROCK1 and ROCK2; Protein imprinting(Western blot) method to detect p-MLC protein expression in gastric tissue.Results:1 Compared with the general condition of ratsNormal group rats have good diet and nourished, responsive, fur burnish, weight gradually increased; Other groups of rats showed polydipsia, polyphagia, polyuria symptoms of poor nutritional status, unresponsive, lethargy, dull color.Two rats died in model group during the experiment. The rats in traditional Chinese medicine group were in better condition than these in diabetic model group.2 Comparison of blood glucose and body weight of ratsCompared with the normal control group, the body weight of model group and Chinese medicine group rats were obviously lower than the normal weight group rats(P<0.05);compared with the model group, Chinese medicine group rats were significantly higher than the normal group(P<0.05). Compared with the normal control group, model group and Chinese medicine group rats blood glucose was significantly higher than the normal group(P<0.05); compared with the model group, Chinese medicine group rats blood glucose was obviously lower than the model group(P< 0.05).3 Comparison of rat gastric emptying rateCompared with the normal control group, model group, Chinese medicine group rats gastric emptying rate decreased significantly(P<0.05); compared with the model group, Chinese medicine group rats gastric emptying rate were significantly higher(P<0.05)4 The results of HE stainingNormal control group rat gastric mucosa, mucosa hierarchical structure clear, neatly arranged glands, rich in blood vessels, smooth muscle cells fusiform nuclei rules, nuclear membrane smooth, dense body cells obviously, no cell vacuoles;model group of rat gastric mucosa is less clear, glandular disorders, reducing the number of vascular smooth muscle cells become smaller, some cells depression, irregular nucleus, the nucleus folds, dense body cells decreased,there vacuolar degeneration, basal membrane thickening;traditional Chinese medicine group the rat gastric mucosa is complete, mucosal structure more clearly, the glands were arranged in order, the number of rich blood vessels, stromal cells.5 Expression of gastric tissue of rats with Rho A m RNAã€ROCK1 m RNA and ROCK2 m RNACompared with normal control group, model group and Chinese traditional medicine group rats stomach tissues Rho Am RNA expression level decreased significantly(P<0.05);Compared with model group,Chinese medicine group of Rho Am RNA expression increased(P<0.05).Compared with normal control group, model group and Chinese traditional medicine group rats stomach tissues ROCK1 m RNA expression level decreased significantly(P<0.05);Compared with model group, Chinese medicine group ROCK1 m RNA expression increased(P<0.05).Compared with normal control group, model group and Chinese traditional medicine group rats stomach tissues ROCK2 m RNA expression level decreased significantly(P<0.05);Compared with model group, Chinese medicine group of ROCK2 m RNA expression were higher(P<0.05).6 Expression of gastric tissue of rats with p-MLC proteinCompared with normal control group, model group and Chinese traditional medicine group rats stomach tissues p-MLC protein expression level decreased significantly(P<0.05);Compared with model group, Chinese medicine group p- MLC protein expression increased(P<0.05).Conclusions:1 Diabetic gastroparesis delayed gastric emptying in rats obvious;Blood circulation meridians of Chinese medicine can significantly improve the rate of gastric emptying in diabetic gastroparesis rats, diabetic gastroparesis have a good therapeutic effect.2 Diabetic gastroparesis rat have abnormal gastric smooth muscle contraction,may be associated with reduced Rho A/ROCK signaling pathways.3 The herbs of activating blood circulation and dissolving stasis medicine may increase rat diabetic gastroparesis Rho A/ROCK signaling pathway in gastric smooth muscle lesions improve, promote gastrointestinal smooth muscle contraction, there by improving the rate of gastric emptying. |
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