| Objective:Acute Radiation Enterntis(ARE)rat model was established by radiation induction.The therapeutic effect of Xiayuxue Decoction on ARE rat model was observed and its mechanism was preliminarily explored.Based on this study,the components of Xiayuxue Decoction were analyzed,and the therapeutic effect of Rhein on ARE rat model and its molecular mechanism were further explored.To provide theoretical and experimental basis for clinical application of Xiayuxue Decoction and its effective components in the treatment of ARE.Methods:Thirty male rats were randomly divided into three groups:group A,normal control group(n=10);group B,normal saline control group(n=10);group C,Xiayuxue Decoction group(n=10).Rat models of acute radiation enteritis were established in group B and group C by single local irradiation of 10 Gy high-energy medical X-ray.Beginning from the 1st day after irradiation,except for the normal control group,group B and group C were intervened with saline and Xiayuxue Decoction respectively.The rats were gavaged for 7 consecutive days.The general conditions of body weight attenuation percentage,fecal traits,fecal blood and autonomous activity were observed daily.Histopathological changes of ileum and colon were observed by HE staining;serum levels of inflammatory related factors such as tumor necrosis factor-alpha(TNF-alpha),interleukin-1(IL-1)and interleukin-6(IL-6)were measured by enzyme-linked immunosorbent assay(ELISA);and intestinal inflammatory factors such as TNF-alpha,IL-1 and I were measured by immunohistochemistry.The expression of L-6.The active ingredients and contents of Xiayuxue Decoction were analyzed by high performance liquid chromatography(HPLC).Fifty male rats were randomly divided into five groups:group A normal control group(n=10),group B saline control group(n=10),group C Rhein group(n=30),group C C1:rhein(9ug/100g)group(n=10),and C2:rhein group.(18ug/100g group(n=10)and C3:rhein group(36ug/100g)(n=10).The same method was used to establish the model and administer the drug.The disease activity index(DAI)was scored and the histopathological changes of ileum and colon were observed by HE staining.The expressions of TNF-a,IL-1 and IL-6 in intestinal tissue of the rat model were determined by immunohistochemistry.The levels of carbon monoxide(NO)and superoxide(SO)in serum of the rat model were measured by enzyme-linked immunosorbent assay(ELISA).The contents of chemical dismutase(SOD),glutathione(GSH)and malondialdehyde(MDA)were measured.The expression of PPAR-gamma and NF-kB in intestinal tissue of rat model was detected by Western-blot.Result:1.The rat model of acute radiation enteritis was successfully established by irradiating the abdomen with medical X-ray linear accelerator.2.After the successful establishment of the model,compared with the normal control group,the rats in each irradiation group showed different changes in body weight,fecal characteristics and autonomous activities.The general situation and defecation situation of the Xiayuxue Decoction treatment group were better than that of the saline group.3.HE staining was used to observe the histopathological changes of ileum and colon in rats.After irradiation,the ileum and colon mucosa of rats in Xiayuxue decoction treatment group were basically intact and inflammation was mild,while the intestinal mucosa of rats in saline group showed necrosis,exfoliation,gland atrophy and severe inflammation.4.Enzyme-linked immunosorbent assay(ELISA)was used to determine the levels of inflammatory related factors in serum of rat model.The results showed that the levels of TNF-alpha in serum of Xiayuxue Decoction group were significantly lower than that of saline group(P<0.01).The contents of IL-1 and IL-6 decreased more significantly(P<0.01).5.Immunohistochemical assay was used to determine the levels of inflammatory related factors in intestinal tract of rats.The results showed that the positive rate of TNF-alpha in ileum of rats treated with Xiayuxue Decoction was significantly lower than that in saline group(P<0.01);the positive rate of IL-1 was significantly different(P<0.05);the positive rate of IL-6 was significantly different(P<0.05).Compared with saline group,the positive rate of TNF-alpha in colon tissue of rats in Xiayuxue Decoction group was significantly lower(P<0.01),the positive rate of IL-1 was significantly different(P<0.01),and the positive rate of IL-6 was significantly different(P<0.05).6."Xiayuxue Decoction"contains the following components:amygdalin,uracil,rhein and emodin.7.After successful modeling,compared with the normal control group,the rats in each irradiation group showed different changes in body weight,fecal characteristics and autonomous activities.The general situation and defecation situation in the Rhein treatment group were better than that in the saline group.8.HE staining was used to observe the histopathological changes of ileum and colon in rats.After irradiation,the intestinal tissue of rats in saline group showed mucosal destruction,villus exfoliation and recess atrophy.The coagulation necrosis of villus and intestinal mubosa of rats in Rhein treatment group was less and the inflammation was milder.Among the three subgroups of Rhein treatment group(C1,C2,C3),intestinal injury in C2 group was lighter than that in C1 group(P<0.05),and intestinal injury in C3 group was lighter than that in C2 group(P<0.05).9.Enzyme-linked immunosorbent assay(ELISA)was used to determine the contents of NO,SOD,GSH and MDA in rat model serum.The results showed that compared with the normal control group,the levels of NO and MDA in rat model serum of each irradiation group increased(P<0.01),the activities of SOD and GSH decreased(P<0.01),and the contents of NO and MDA in rat model serum of Rhein treatment group decreased significantly(P<0.01).The activities of SOD and GSH were higher than those of saline group(P<0.01).Among the three subgroups(C1,C2,C3)of Rhein treatment group,the contents of NO and MDA in C3 group were lower than those in C1 and C2 group(P<0.01),and the activities of SOD and GSH in C3 group were higher than those in C1 and C2 group(P<0.01).10.Immunohistochemical assay was used to determine the levels of inflammatory factors in intestinal tract of rats.The results showed that the expressions of TNF-a,IL-1 and IL-6 in intestinal tissues of rats in each irradiation group were higher than those in normal control group(P<0.05),and the expressions of TNF-a,IL-1 and IL-6 in intestinal tissues of rats in saline group were significantly higher than those in Rhein treatment group(P<0.05),Rhein treatment group(P<0.05).Among the three subgroups(C1,C2,C3),the expression of TNF-a,IL-1 and IL-6 in intestinal tract of C3 group was lower than that of C1 and C2 group(P<0.05).There was no significant difference in the expression of TNF-a,IL-1 and IL-6 in intestinal tract between C1 and C2 group(P>0.05).11.Western-blot was used to detect the expression of PPAR-gamma/NF-kB in intestinal tissue of rat model.The experimental results showed that the expression of PPAR-gamma in the irradiated group was significantly lower than that in the normal control group(P<0.01).The expression of PPAR-gamma in the Rhein treatment group was significantly higher than that in the NS group(P<0.01).There was no significant difference among the three subgroups in the Rhein treatment group(P>0.05)?On the contrary,the expression of NF-kB was significantly increased in all irradiation groups(P<0.01).The expression of NF-kB in Rhein treatment group was significantly lower than that in NS group(P<0.01).In the three subgroups of Rhein treatment group,the expression of NF-kB decreased with the increase of Rhein concentration(P<0.01).Conclusion:1."Xiayuxue Decoction"can effectively alleviate the symptoms of acute radiation enteritis in rats,such as emaciation,diarrhea,fecal blood,etc.2."Xiayuxue Decoction" has protective effect on intestinal mucosa of rats with acute radiation enteritis.It can effectively reduce the concentration of inflammatory related factors TNF-a,IL-1 and IL-6 in serum arid intestinal tissue of rats.3.Preliminary determination of "Xiayuxue Decoction" contains the following components:rhein,amygdalin,uracil and emodin.4.Rhein,an effective component of Xiayuxue Decoction,can effectively alleviate the symptoms of acute radiation enteritis in rats,such as emaciation,diarrhea,stool blood,etc.5.Rhein,an effective component of Xiayuxue Decoction,can reduce the concentration of inflammatory related factors TNF-alpha,IL-1 and IL-6 in intestinal tissues of rat models and play an anti-inflammatory role in acute radiation enteritis.6.Rhein,an effective component of Xiayuxue Decoction,can increase the content of SOD and GSH in serum of rat model and decrease the expression of NO and MDA,thus achieving the anti-oxidative damage of acute radiation enteritis.7.The mechanism of effective ingredient rhein in Xiayuxue Decoction in treating acute radiation enteritis is related to PPAR-gamma/NF-kB pathway. |
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