The Treatment Effect, Mechanism Of Action And Safety Of Tongshuang On Experimental Constipation

ObjectiveStudying the treatment effect,mechanism of action and safety of TongSHuang on experimental constipation to provide a scientific basis for its clinical application.Method1. Curative effect search(1) Modeling. With diphenoxylate 0.2 ml / mouse established constipation model.(2) The time of first defecation and moisture content. Mice in each group recorded the time of first defecation and calculate fecal water content.(3) The rate of ink. To use ink as indicator, the ink in each group were measured in mice small and large intestine of the advance distance, calculate the rate of ink.2. Mechanism research methods(1) Bowel movement in vitro. Preparing rat intestinal muscle specimens, PCLAB-UE record curve of normal condition and TongSHuang with different concentrations works intestinal smooth muscle in vitro.(2) Jejunum movement. Rabbits implanted with jejunal water bag method, duodenal administration, PCLAB-UE record the jejunum motion curve with normal physiological conditions, and different concentrations of TongSHuang.(3) Ileum movement. Intestine with the suspension method, PCLAB-UE record the ileum motion curve with normal physiological conditions, and different concentrations of TongSHuang.(4) Intestinal fluid volume changes. The rabbit colon and ileum were ligated, injected with saline, Maren Wan liquid, cisapride solution and the groups of TongSHuang each 2ml, after 2h measured amount of fluid within the bowel, and observe the intestinal wall and intestinal swelling mucosal hyperemia.(5) Colonic myoelectricity. Rat constipation model were established by Diphenoxylate, with PCLAB-UE determination of rat colonic myoelectricity.3. Safety test methods(1) Take ink test mice produced large intestine intestines were HE stained paraffin sections (2) Acute toxicity.Result1. Curative effect results(1)Modeling results show that the first row of blank group were black particles will be in 157.6±10.49 min, fecal water content of 32.79±1.829%, constipation model group 363.9±12.851 min, water content of 29.649±1.2307%, two groups were significantly different (p <0.01), the model was successful.(2) TongSHuang low-dose group were ranked first black particles at 135.4±8.76 min, fecal water content was 38.46±1.374%, compared with model group, two groups were significantly different (P <0.01); dose groups in the first row of the black particles at 89.70±9.00 min, fecal water content of 41.00±3.456%, the high dose group at 73.06±7.49 min, water content of 43.42±1.189%, cisapride group 120.0±13.11 min, moisture content of 35.04±1.554%, Maren Wan Group 111.2±13.86 min, water content 39.78±1.564%, dose and high doses compare with Maren Wan and cisapride groups were significantly different.(3) In the small intestine and large intestine of ink test, the model group of ink were 34.98±5.26% and 11.21±4.65%, low dose group 64.79±5.08% and 24.44±4.87%, the dose group 78.31±4.50% and 36.63±6.56%, high dose group 87.03±3.80% and 56.49±7.97%, cisapride group 63.00±4.72% and 28.32±7.77%, Maren Wan group 63.59±6.80% and 36.11±11.19%. Low-dose group compare with model group were significantly different (P <0.01); dose and high dose group and Maren Wan, cisapride group was significantly different (P <0.01).2. Mechanism study result(1) Control group of isolated small intestinal movement amplitude 184.66±6.19 mg, TongSHuang low dose group 192.72±16.02 mg, and the control group significantly different (P <0.05); dose group 245.75±17.92 mg, high dose of 272.61±20.55 mg, middle and high dose group compared with the control group were significantly different (P <0.01).(2) After adding ventilated granules, compared with the control group, low dose group TongSHuang increase in pressure of small intestine 35.0±2.93%, an increase in the dose of 70.94±12.73% high-dose group increased by 159.65±27.77%.(3) Suspension for trial, after adding ventilated granules, compared with the control group, low dose group TongSHuang increase in pressure of small intestine 0.49±0.02%, the dose increased 0.94±0.38%, an increase of high-dose group 0.98±0.04%.(4) Rabbit intestine drug delivery trials, three doses TongSHuang into the ileum and colon segment intestinal fluid volume was 2.38±0.13 ml, 3.12±0.54 ml, 4.10±0.91 ml and 2.28±0.10 ml, 2.90±0.22 ml, 3.85±0.08 ml compared with control group 1.14±0.65 ml, 1.00±0.08 ml in each group were fluid volume increases, the difference was significant (P<0.01); by cisapride and Maren Wan role of intestinal mucosa of severe congestion and swelling , while the role of bowel TongSHuang with the control group showed no changs.(5) Normal mice rhythm of 13.6±0.29 times / min, range 0.17±0.01 mv. Slow wave frequency group 6.92±0.35 times / min, range 0.47±0.03 mv, TongSHuang treated frequency 9.03±0.89 times / min, amplitude 0.3±0.02 mv, group differences between the two data were significantly (P < 0.05); rhythm of rats to speed up the frequency 31.43±2.78 times / min, range 0.28±0.25 mv, after treatment by ventilation granules slow wave frequency 20.35±1.13 times / min, amplitude of 0.20±0.01 mv, group two Data significantly different (P <0.05).3. Safty research result(1) Microscope HE staining, the role of TongSHuang compared with the control segments of intestine, colonic mucosa structure intact epithelium without stripping, no abnormal intrinsic glands, stromal-free congestion, edema, inflammatory cell infiltration, myenteric plexus, there were no obvious changes in muscle cells; cell structural integrity, structural integrity of villi; cell loss was not found, cell shrinkage or swelling does not appeard,also does not appear the nucleus stain, increase in the size.(2) Largest delivery volume test results indicate that the maximum delivery capacity of 60 g·kg^-1·d^-1, conversion of crude drug content of 289.6 g·kg^-1·d^-1.Conclusion1. TongSHuang can solve the experimental constipation problem. TongSHuang on both colon and small intestine contents were significantly advanced the promotion of the role and performance of enhanced with increasing concentration.2. TongSHuang significantly increased rate of intestinal peristalsis, and promote intestinal absorption of nutrients and accelerate the fecal excretion.3. TongSHuang can increase intestinal fluid volume in order to soften the stool, and promote colon emptying, and stimulating small.4. TongSHuang can regulate abnormal colonic muscle disorder, so that abnormal colonic slow wave frequency and amplitude of recovery, so that the colon became normal electrical activity.5. TongSHuang will not be used due to the cathartic colon and the role of enteric nervous system damage, it will not appear harm effect on the body, non-toxic to the body, safe, reasonable, with further value of the study.