Evaluation The Regeneration Effect Of Liang Xue Yu Chang Decoction On Intestinal Epithelium Of Rats With Radiation Enteritis

ObjectiveRadiation therapy is a kind of important method for the treatment ofabdominal pelvic tumor, and radioactive enteritis is common complication ofradiation. Acute radioactive enteritis mainly had bowel mucosal epitheliumintegrity destruction, and disorders of epithelial cells from diarrhea symptomssuch as bleeding; chronic radiation enteritis caused by chronic ulcer,hemorrhage, fistula formation, and obstruction which are the results of thebowel wall fibrosis.At present the treatment method of radioactive enteritis include:(1)intestinal mucosa protector, but the total effect is not confirmed;(2) parenteralnutrition: glutamine and parenteral nutrition have certain curative effect, but thecost is high, and there is some complications such as catheter sepsis, liverdamage, the metabolic disorder;(3) hyperbaric oxygen therapy: can increasethe damage of oxygen supply in the intestinal tract, promote damage repair, buthyperbaric oxygen therapy need special large equipment;(4) the endoscopicand surgical treatment: endoscopic therapy suits local haemostatic effect,patients with serious symptoms will only accept surgery, cut off the diseasedbowel;(5) traditional Chinese medicine, Chinese medicine as one of China’straditional medicine treasure, has unique advantages in the gastrointestinal tractdisease’s treatment, there are few studies reported that traditional Chinesemedicine has a good curative effect on radioactive enteritis,can effectivelyrelieve symptoms, promote the body recovery. But most limited to clinicalcurative effect observation, lack of histology, cytology, molecular level research.This topic is based on the previous work, and built rat model of bowel damage, from the histological, cytology, molecular level, multi-levels study thetherapeutic effect of Liang soup on damaged bowel, and to explore themechanism.Methods1、The preparation and group radioactive enteritis rat models140SD rats were randomly divided into4groups. Group A(n=20) was thecontrol group, Group B(n=40)、 Group C(n=40) and Group D(n=40) wererespectively underwent whole abdominal irradiation with X-rays at an singledose of5Gy、10Gy and15Gy, to establish the radioactive enteritis rat models.2、The treatment of the radioactive enteritis rat models.After the completion of radiation enteritis model, group B、group C andGroup D were randomly divided into two groups, group B1、group C1and groupD1after irradiation were administrated by i.g. with distilled water, group B2、group C2and group D2after irradiation were with Liangxueyuchang Decoction.3、Detect the epithelial cell proliferation trough Brdu methodBefore to be sacrificed,rat in each group were injected Brdu solution,andthen prepar paraffin sections follow the same methods,detect the Brdu positivecells through immunohistochemical method.4、Detection the PCNA and Ki67expression in intestinalepithelium cells of rats in each groupStain the PCNA and Ki67in paraffin sections using SP immunohistochemical kit,and follow the structions of the kit.5、Detection the inflammatory factors expression in intestinaltissue of rats in each groupPut the rats to death, and take total RNA from the tissue, and detected theexpression changes of IL-6、IL-8、IL-1、TNF-ɑ、IL-4、IL-10with real-time PCR. Results1、Test the proliferation ability of intestinal mucosa epithelialcellsIn normal rats group, there are many PCNA and ki67positive cells, inmodel rats group, epithelial cells of villi with PCNA and ki67express significantlyreduced, treated with Liangxueyuchang Decoction, PCNA and ki67positiveepithelial cells of villi in a model group significantly increased, but less thannormal group.2、Detection the number of epithelial cell of bowel mucosa in Sphase through Brdu methodThere are many Brdu marked positive cells (S phase cells) in villi,and inthe model group, the Brdu mark positive cells reduced significantly, and themodel group treated with Liangxueyuchang Decoction, the Brdu markedpositive cells increased significantly compared with the untreated modelgroup.3、 Detection the inflammatory factors expression in intestinaltissue of rats in each groupIn the intestinal tissue of normal rats, inflammation factors such as IL-6、IL-8、IL-1、TNF-ɑ expression is very little, in model rats, the expression of thevarious factors are significantly increased, treated with LiangxueyuchangDecoction, the expression of IL-4, IL-10, in intestinal tissue of rat modelsdropped significantly, but higher than normal rats.ConclusionLiangxueyuchang Decoction can accelerate recovery of injured intestinalmucosal epithelium, promote the proliferation of epithelial cell of bowel mucosa,reduce the inflammatory factor expression in damaged bowel tissue, to maintainstability of the structure and function.