| [Background]Gastroesophageal reflux disease (GERD) refers to the gastric contents reflux into the esophagus and cause discomfort symptoms and (or) a group of complications disease.It has three types, non-erosive reflux disease(NERD), reflux esophagitis(RE), Barrett’s esophagus(BE).GERD is a common and frequently-occurring disease and the morbidity is increasing.The treatment of Modern Medicine of GERD include of inhibition of gastric acid, promoting gastrointestinal motility, protecting gastric mucosa or a surgical operation. Traditional Chinese medicine has certain advantages in the treatment of GERD.This research is to study the mechanism of Tongjiang Granule on reflux esophagitis,which using a rat model of reflux esophagitis with Syndrome of disharmony between liver and stomach.[Purpose]After discussing the progress of the treatment of Modern Western Mdeicine,analyzing its advantages and problems,to provide references for scientific research.Conducted Tongjiang Granule pharmacodynamic experiments to clarify its pharmacological effects;to give different doses of Tongjiang Granule to a rat model of reflux esophagitis with Syndrome of disharmony between liver and stomach, to study the mechanism of Tongjiang Granule on reflux esophagitis.[Research]Research I Pharmacodynamics of Tongjiang GranuleObjective:The effect of the Tongjiang Granule on gastrointestinal motility and its anti-inflammatory and analgesic effects.Materials and methods:In reflux esophagitis rats on gastric emptying and small intestinal propulsion experiments to observe the effects of the Tongjiang Granule on the gastrointestinal motility of rats model by acetic acid writhing method and xylene induced mouse ear swelling test to observe whether the anti-inflammatory and analgesic effects of Tongjiang Granule.Results:(1) In experimental of model rat gastric emptying, high dose group of and middle dose group, the gastric residual rate were higher than those of the sham operation group (P< 0.05), middle dose group, mosapride group, middle dose combined omeprazole group gastric residual rate was lower than that of model group (P< 0.01), low dose group, omeprazole group gastric residual rate was also lower than that of model group (P< 0.05), suggesting that Tongjiang Granule middle dose〠low dose and omeprazole, mosapride has positive effect on Rats’ gastric emptying movement;(2) In model rat small intestine propulsive motility experiments, high dose group, middle dose group, omeprazole group, middle dose combined omeprazole group, the intestinal propulsion rate were higher than those in the model group (P< 0.05 or P< 0.01), suggesting that high and middle dose could promote the small intestine in rats.(3) In the mice acetic acid torsion test, compared with normal group.aspirin groups high dose group and middle dose group were statistically significant (P< 0.01 or P< 0.05), suggesting that high doseã€middle doseã€aspirin has a certain analgesic effect;(4) in xylene induced mouse ear swelling test, middle dose group and aspirin group ear swelling degrees below normal group (P< 0.05, P< 0.01). suggesting that Tongjiang Granule have a certain anti-inflammatory effect.Research II Study on reflux esophagitis between the liver and stomach syndrome rat model of gastrointestinal motility mechanism of Tongjiang GranuleObjective:By observing the intervention Tongjiang Granule on reflux esophagitis liver stomach disharmony of model rats, from the serum gastrointestinal hormones, calmodulin and myosin light chain kinase of different molecular biology level detection, trying to regulate the mechanism of gastrointestinal motility.Materials and methods:Side to side anastomosis of the esophagus and duodenum, superimposed on the tail stimulation, established the reflux esophagitis rat model of liver stomach disharmony. Experimental animals were divided into sham operation group, model group, high dose group, middle dose group, low dose group, omeprazole group, mosapride group, middle dose combined omeprazole group. At 9 o’clock in the morning, the sham operation group and the model group were given distilled water respectively, the treatment group were given 1ml/100g rats by body weight for 21 days. In the process, the body weight, food intake, drinking water quantity, grasping force, hair color, activity and so on were observed and recorded. After the treatment, the abdominal aorta was taken from the abdominal aorta, the esophageal tissue was taken and the HE staining was used to observe the pathological changes of the esophagus, the relative expression of Cam and MLCK protein was detected by blot Western blotting, the relative expression of MLCK and Cam in esophageal tissues were detected by immunohistochemistry,the relative expression of MLCK and mRNA Cam in esophageal tissues by Time PCR Real.ResultsGeneral situationAfter three weeks, the high doseã€middle dose group and omeprazole group, middle dose combined omeprazole group relative to the weight of the model group increased (P< 0.05); high dose group, omeprazole group, middle dose combined omeprazole group compared with the model group, the food intake increased (P< 0.05); omeprazole group, mosapride group compared with the model group, the food intake increased (P< 0.05); high dose group, mosapride group, middle dose combined omeprazole group and the model group, the comparison, the grasping force difference has statistical significance (P< 0.05)Serological detectionMotilin content, and compared with sham operation group, high dose group, middle dose combined omeprazole group, the serum motilin content was higher than that of sham operation group (P< 0.05); compared with the model group, high dose group and Mosapride will benefit group, serum motilin content was higher than that in the model group (P< 0.05).Gastrin content, compared with the sham operation group, model group, the serum gastrin content lower than sham operation group (P< 0.01). The rest of the group and the sham operation groups were no statistical significance (P> 0.05); compared with the model group, high, medium and low dose group, middle dose combined omeprazole group and sham operation group, the serum gastrin content was higher than that in the model group (P< 0.01) and Mosapride group, serum gastrin content was higher than that in the model group (P< 0.05), omeprazole group and the model group without statistical significance (P> 0.05).The content of CCK, compared with the sham operation group, high dose group and model group, the serum content of CCK were lower than in the sham operation group (P< 0.01); compared with the model group, mosapride will group and middle dose combined omeprazole group, the serum content of CCK were higher than in the model group (P< 0.01).Vasoactive intestinal peptide, and compared with sham operation group, high dose group, the serum levels of vasoactive intestinal peptide content lower than sham operation group (P< 0.05), dose, dose combined omeprazole group and model group, the serum levels of vasoactive intestinal peptide content higher than in the sham operation group (P< 0.05); compared with the model group, the content of high dose group and omeprazole group, the serum levels of vasoactive intestinal peptide is lower than the model group (P< 0.01).Nitric oxide content, and compared with sham operation group, reduced particle dose group, low dose group and middle dose combination content of serum nitric oxide in the omeprazole group was higher than that in sham operation group (P< 0.05, P< 0.01); compared with the model group, drop particle dose group, low dose group, mosapride group, middle dose combined omeprazole group, the serum nitric oxide content were higher than those in the model group (P< 0.01).Western blotThe relative expression of MLCK:the model group (P< 0.05), high dose group (P< 0.01), low dose group (P< 0.05) and omeprazole group (P< 0.05) and Mosapride group (P< 0.01) group and sham operation compared MLCK relative expression difference was statistically significant; except the omeprazole group and sham operation group, the rest of the group and the model group compared the MLCK relative expression the difference was statistically significant (P< 0.01).The relative expression of CaM:High dose group, mosapride group and sham operation group compared the difference was statistically significant (P< 0.01); high dose (P< 0.01), mosapride group (P< 0.01), middle dose (P< 0.05), middle dose combined omeprazole group (P< 0.05) compared with the model group, the difference has statistical significance.Immunohistochemical detectionExpression of MLCK have certain difference, the model group showed negative results and no positive brown granules; omeprazole group were false positive results, other groups showed positive or weakly positive results, and model group differences.Expression of CaM in each group had a certain difference, model group showed false positive results; the other groups showed positive or weak positive results, there is a difference between the model group and the model group.Real Time PCRCam mRNA relative expression, mosapride group (P< 0.01), high dose group (P < 0.05), medium dose and omeprazole group (P< 0.05) differences with the sham operation group and the model group have statistical significance.MLCK mRNA relative expression, mosapride group (P< 0.01), middle dose combined omeprazole group (P< 0.05) compared with the sham operation group, there was significant difference; high dose (P< 0.05). middle dose group (P< 0.05) and omeprazole group (P< 0.05) and Mosapride group (P< 0.01), middle dose combined omeprazole group (P< 0.01) model group compared the differences have statistical significance.[Conclusion]1 Tongjiang Granule has the effect of promoting gastric emptying and small intestine propulsion and anti inflammation and analgesia.2 Tongjiang Granule to promote gastrointestinal motility may by regulating gastrointestinal hormones, the promotion or inhibition of motilin, gastrin, cholecystokinin, vasoactive intestinal peptide, nitric oxide secretion and release, so as to adjust the esophageal and gastrointestinal movement.3 Tongjiang Granule can affect the expression of CaM, MLCK protein and gene in esophageal tissue. |
