A Study To The Influence Of The 3-Leg Points In Jin’s 3-Needle Technique To Cajal Cells Repair After Colon Anastomosis, A Perspective Of NO Signal

Surgical postoperative gastrointestinal dysmotility is one of the most common clinical syndrome symbolized by tract symptoms. Abdominal pain, nausea, vomiting, exhaust bowel symptoms ect are mainly caused by weakness of gastrointestinal movement and difficulty in passing through gastrointestinal tract after operation, especially after abdominal surgery, as a result they affect recovery badly in later period. Many Gastrointestinal dysmotility disorders are affected by many factors in operation. Because its pathogenesis is not fully understood, there is no effective modern clinical treatment. As postoperative gastrointestinal dysmotility are concerned by more and more people, it became a research hot spot. Based on treatment characteristics of TCM whose treatment varies from person to person and is based on syndrome differentiation, it has obvious curative effect in adjusting gastrointestinal function after surgery. And acupuncture and moxibustion in TCM, owing to that there is no postoperative drug stimulation on gastrointestinal tract and its high security, are gaining more and more attention from surgeons both home and abroad.ObjectiveThe-3-Leg points in Jin’s 3-Needle Technique is one of the most commonly used acupuncture point group, which is concluded and founded by professor JinRui of Guangzhou University of Chinese Medicine. It consists of zusanli, sanyinjiao, taichong acupoint, which can invigorate spleen and sooth liver when receiving acupuncture. When it is applied to patients after abdominal surgery, it shortens the exhausting and defecating time significantly in clinical treatment. On such basis, the author takes the-3-Leg points in Jin’ s 3-Needle Technique to acupuncture SD rats that have been given colon anastomosis successfully and takes down the time of first defecation after operation. The author also observes the propulsive rate of small intestine, amount of nitric oxide(NO), activity of nitric oxide synthase(NOS) and expression Cajal cells c-kit in tissues at certain time. By following NO gas signal, the author tries to understand how the-3-Leg points in Jin’s 3-Needle Technique improves rats’ gastrointestinal movement, and at the same time, by comparing concerning indicators taken from rats that are treated with different acupuncture time, it provides reference of the right timing for such treatment in clinical.MethodDocument research mainly summarizes the methods of research to the etiology, pathology and treatment to postoperative gastrointestinal dysmotility in modern medicine, and the understanding, analysis, as well as major treatments in modem and traditional Chinese medicine.Colonic anastomosis is done to rats to make gastrointestinal dysmotility model, and the rats are, according to the requirements of experiment, divided into normal group,3-day model group,3-day acupuncture group,5-day model group,5-day acupuncture group,10-day model group and 10-day acupuncture group, and ten rats in each group, seven groups in total. Acupuncture is given at every evening, and the first defecation time is recorded. After a specified period of time,1 ml of ink is given by gavage to all rats that will be executed half an hour later, after which the propulsive rate of small intestine is taken down by the author, and the NO, NOS and c-kit in the 1 cm long colon tissue near which colonic anastomosis is carried out are examined simultaneously.Result1. Results of first defecation after operation:the first defecation time of acuPuncture group is shorter than that of model group(P＜0.01);2.Results of propulsive rate of small intestine:compared with normal group (P＜0.01), the propulsive rate of small intestine of 3-day model group,5-day model group,10-day model group,3-day acupuncture group are lower, while no statistical significance is found in 5-day acupuncture group,10-day acupuncture group when compared with normal group(P＞0.05); the small intestine propulsion rate of 3-day acupuncture group is less than that 5 and 10-day acupuncture group (P＜0.01) and there is no statistical difference between 5-day and 10-day acupuncture group when all acupuncture group are compared with each other. When all model groups are brought into comparison, it finds no statistical significance between 3-day and 5-day model group(P＞0.05) and 5-day model group has less propulsive rates of small intestine than 10 days model group(P＜0.05); When acupuncture group is compared with model group, no statistical significance exists between 3-day acupuncture group and 10-day model group(P＞0.05).3. Examination results of NO:Nitrate reductive enzymatic is adopted to measure the amount of NO content in colon tissue of different groups. ComPared with normal group, model groups and acupuncture group have higher amount of NO (P＜0.05); 3-day model group has more NO than that of 5 and 10-day model group (P＜0.05), and 5-day model group gets more NO than 10-day model group (P＜0.01) when model groups are compared with each other; More NO is examined in 3-day acupuncture group than 5 and 10-day acupuncture group(P＜0.01); and similarly, more NO in 5-day acupuncture group than in 10-day acupuncture group(P＜0.01), when comparing acupuncture groups with each other; In comparison of model and acupuncture groups, there is no statistical significance between 3-day acupuncture group and 10-day model group concerning the amount of N0(P＞0.05), but in 5-day acupuncture and 10-day model groups (P＜0.05).4. NOS enzyme activity test results:the L-arginine (L-Arg) is adopted to test activity of nitric oxide synthase (NOS) enzyme, which are consist of nitric oxide synthetase (cNOS) and inducible nitric oxide synthetase (iNOS) regarding that whether it relies on intracellular calcium physiological regulation. Comparison between groups of rats NOS (TNOS, iNOS, cNOS) enzyme activity, compared with normal group, each group TNOS、iNOS enzyme activities were higher than normal group, cNOS were lower than normal group, and all those difference is of statisticalal significance (P＜0.05); With the time increasing, TNOS and iNOS enzyme activity comparison between model groups,, It is found that 3-day group is higher than that of 5-day group,5-day group is higher than 10-day group, and difference is statistically significant(P＜0.05), but the cNOS enzyme activity comparison between model groups,3-day group is lower than that of 5-day group,5-day group is lower than 10-day group (P＜0.05), and the difference is statistically significant (P＜0.05); TNOS and iNOS enzyme activity comparison between acupuncture groups, It is found that 3-day group is higher than that of 5-day group,5-day group is higher than 10-day group, and difference is statistically significant(P＜0.05), and the cNOS enzyme activity comparison between acupuncture groups,,5-day group is lower than 10-day group(P＜0.01), but no statistical difference is found between 3-day group and 5-day group(P＞0.05); When each model group compared with each acupuncture group, the activity of cNOS、iNOS enzyme of 3-day acupuncture group has no statistical difference with 10-day model group (P>0.05), but the activity of TNOS enzyme has no difference between 5-day acuPuncture group and 10-day model group(P>0.05).5. Results of the average absorption of c-kit:average absorption of 3-day model group,5-day model group,10-day model group and 3-day acupuncture group is less than that of normal group (P＜0.01); that of 3-day model group is lower than 5-day model group and 5-day model group lower than 10-day model group and it is statistically different (P＜0.05); for acupuncture groups,3-day acupuncture group has less absorption than that of 5-day and 10-day acupuncture groups(P＜0.01), but absorption of 5-day acupuncture group is not statistically different from that of 10-day acupuncture group(P＞0.05); for the difference between acupuncture and model group, the difference between 3-day acupuncture group 10-day model group is not statistically significant(P＞0.05), but vice versa between 5-day acupuncture group and 10-day model group(P＜0.05).Conclusion:1. "The-3-Leg points" intervention in early period after surgery can obviously shorten the first defecation time.2. (1)postoperative acupuncture improves the propulsive rate of small intestine and strengthens the function of gastrointestinal activities;(2) There is a sellf-healing tendency in rats of colonic anastomosis and as time goes by, the small intestine propulsive rate improves obviously. However, compared with acupuncture group, the model group recover more slowly and when acupuncture is given in the 5th day, they are not different to normal group significantly in terms of intestine propulsion rate.3. Colonic anastomosis damages the gastrointestinal pacemaker Cajal cells,whose recovery is related with the NO level in its surrounding closely. Such a surgery leads to over level of NO. The acuPuncture intervention given in early period makes inhibition to NO level, and as the acupuncture time extends, the NO content in the colon anastomotic tissue drops;4. Generation of NO is related to the NOS enzyme activity. The stress conditions caused by surgery promotes the synthesis of iNOS (inducible nitric oxide synthetase) and cNOS (constitutive nitric oxide synthetase) expression in a short period of time. The acupuncture suppress the enzyme activity of iNOS but accelerate the cNOS enzyme activity. As time passes by, TNOS and iNOS enzyme are less active, but the change in iNOS enzyme activity in acupuncture group is not that obvious as that of model group, and the cNOS is more than that of model group and it is not recover to normal level by the 10th day.5. c-kit protein is a symbol of the development and function performance of Cajal cells. Acupuncture reduces the damage by anastomosis colon to cajal cellsf, and promotes the expression of c-kit protein and facilitate postoperative Cajal cells regeneration and functional recovery.6. Acupuncture after colonic anastomosis to rats relieves damage to ICCc cells and facilitate the regeneration of Cajal cells by promoting the expression of cNOS enzyme and suppress the iNOS enzyme acitivity, and regulate the production of the NO collectively, thus help with the recovery of gastrointestinal activity.7. Colonic anastomosis does serious damage to intestinal, which takes rather a long time to recover, it reduces gastrointestinal reaction after a short time of acupuncture and shortens the first defecation time after operation, but it requires a longer acupuncture time to form a favorite micro environment for the recovery of Cajal cells, so as to improve the gastrointestinal movement function and promote the overall recovery of patients.