The Study On The Effects And Mechanisms Of Treatment Of Huaihuasan And Baitouweng Decoction In Rats With Acute Radiation Proctitis

Objective: To observe the curative effect of Huaihuasan combined with Baitouweng decoction on acute radiation proctitis in rats and explore its mechanism.Methods: 40 clean SD rats were randomly divided into blank Control group,Model group,Mesalazine group and Huaihuasan combined with Baitouweng decoction group(BTW),with 10 rats in each group.Except for the Control group,rats in the other three groups were all irradiated with a dose of6 m V-X ray of 20 Gy in the pelvic cavity to make the rat model of acute radiation proctitis.After model formation,the rats were given corresponding drug gavage treatment every day.On the 14 th day after gavage,all the rats were sacrificed to evaluate the general condition of the rats.The pathological changes of the rectal tissue of the rats were observed by HE staining.The changes of intestinal microflora in ARP rats were detected by 16 S r DNA sequencing technology.The protein expression levels of nuclear transcription factor-kapp Ab(NF-κB),phosphorylated nuclear transcription factor-kapp Ab(P-NF-κB)and Toll-like receptor 4(TLR4)in tissues were detected by Western Blot.The protein expression levels of phosphorylated nuclear transcription factor-kappa B(p-NF-κB)and Toll-like receptor 4(TLR4)in tissues were detected by immunohistochemistry(IHC).The levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),intercellular adhesion molecule-1(ICAM-1)and vascular cell adhesion molecule-1(VCAM-1)were determined by enzyme-linked immunosorbent assay(ELISA).The expression of nuclear transcription factor-kappa B(NF-κB)in tissues was detected.All data were analyzed by SPSS 25.0 statistical software,and data results wereexpressed as (?)±s.One-way analysis of variance was used for comparison between multiple groups,and T test was used for comparison between two groups.P< 0.05 was considered statistically significant.Results:1.After irradiation,there was no significant change in the first 2 days in each irradiation group.On the 3rd day,different degrees of food intake,water intake,weight loss,slow reaction,dirty hair color,curling up and less movement,different degrees of loose stools and mucus appeared,and stool frequency increased.The general symptoms of the BTW and Mesalazine groups were milder than those of the Model group.2.He pathological staining section of the rat rectum showed that the epithelial cells in the Model group were sparse,with partial exfoliation,erosion and necrosis.The mucosa and submucosa showed obvious vacuol-like degeneration,increased interstitial fibers and infiltration of a large number of inflammatory cells.In the Mesalazine group and the BTW group,the integrity of rectal epithelial cells was better,the structural changes of glands were less severe,and the morphological structure of rectal mucosa was better,without obvious ulceration,bleeding and inflammatory cell infiltration.3.The 16 Sr DNA gene sequencing of rats’ feces showed that the species abundance of Lactobacillus in the Control group and Mesalazine group was higher than that in the Model group.4.Western Blot results showed that compared with Control group,the expression of TLR4 and P-NF-κB protein in Model group was significantly increased(P<0.01).The protein expressions of TLR4 and P-NF-κB in Model group were significantly higher than those in Mesalazine group and BTW group(P<0.05).5.Immunohistochemical results showed that the yellow staining of cell membrane and cytoplasm was significantly deeper in the Model group than in the Control group,indicating that the expressions of P-NF-κB and TLR4 were significantly higher than in the Control group.The yellow staining of membrane and cytoplasm in Mesalazine group and BTW group was lighter than that in model group and slightly darker than that in Control group,indicating decreased expression of P-NF-κB and TLR4 protein in BTW group.6.ELISA results showed that compared with the Control group,the expressions of TNF-α,IL-6,ICAM-1 and VCAM-1 in serum of rats in the Model group were significantly increased(P<0.01),and the levels of TNF-α,IL-6,ICAM-1 and VCAM-1 in serum of rats in the Mesalazine group and the BTW group were significantly lower than those in the Model group,with a statistical difference(P<0.05).Conclusions:1.Huaihuasan combined with Baitouweng decoction can effectively improve diarrhea,mucinous stools,hematochezia and other symptoms in rats with acute radiation proctitis caused by radiation.2.The mechanism of Huaihuasan combined with Baitouweng decoction in the treatment of acute radiation proctitis in rats is related to the inhibition of TLR4/NF-κB signaling pathway and the reduction of the release of inflammatory factors3.Huaihuasan combined with Baitouweng decoction can improve intestinal flora and increase the abundance of Lactobacillus flora.