| Background:Ciji-Hua'ai-Baosheng ? Foramula(CHB-?-F)is a traditional Chinese medicine prescription for the treatment of tumor patients after the treatment of chemical drugs.The clinical treatment indicated that CHB-?-F can significantly relieve the gastrointestinal toxicity caused by the chemotherapy,reduce the vomiting of the patient,stimulate the appetite of the patient,increase the diet amount and the body weight of the patient,and greatly improve the life state of the tumor patient during the chemotherapy period and after the chemotherapy;However,the mechanism by which CHB-?-F could improve the side effects of chemotherapy on gastrointestinal tract was not clear enough.Aim:To explore the therapeutic effect and mechanism of Ciji-Hua' ai-Baosheng ?Formula on gastrointestinal toxicity induced by chemotherapy in tumor-bearing mice.Methods:A total of 72 healthy Kunming mice of SPF grade were procured.H22 hepatoma cells were inoculated subcutaneously in the right anterior armpit of the mice.After one week of feeding,the tumor blocks visible to the naked eye were developed.The tumor chemotherapy model was established by intraabdominal injection of 5-fluorouracil(5-FU)at a high dose of 200mg/kg.The mice after successful chemotherapy were randomly divided into 6 groups:western medicine control group(5-FU),negative control group(Negative Control),Chinese patent medicine control group(YZXJC),CHB-?-F high dose group(65g/kg),CHB-?-F middle dose group(32.5g/kg)and CHB-?-F low dose group(16.25g/kg).On the second day after the establishment of the model,the mice were given medication for 14 consecutive days,and the body weight and food intake of the mice were measured and recorded on the 7th and 14th day,respectively.The long diameter(1d)and the short diameter(sd)of the tumor were measured and recorded on the 7th and 14th day respectively.At the end of the last drug,fasting water for 12 hours,on the 15th day,the mice were given appropriate amount of anesthetic drugs.After the anesthesia was successful,the mouse eyeball was removed to get blood.The tumor weight was recorded and the tumor inhibition rate was calculated according to the formula.After HE staining,the tumor tissue was dehydrated,embedded and sectioned,and the pathological changes of the tumor tissue were observed and photographed under light microscope.The duodenum and stomach tissues were desiccated,embedded and sectioned,and then stained with HE,the histopathological changes were observed by light microscope.Detection of Leptin,Neuropeptide Y(NPY),Orexin A(OXA),Gastrin(GAS),Ghrelin,Prostaglandin E2(PGE2),Epidermal growth factor(EGF),Motilm(MTL)in serum of mice by Enzyme-linked immunosorbent assay(ELISA);Superoxide dismutase(SOD)activity and Malondialdehyde(MDA)content in jejunum of mice by ELISA;Detection of Proopiomelanocortin(POMC),Orexin Receptor 1(OX1R),NPY,Agouti gene-related protein(AgRP),Leptin Receptor(Ob-R),Cocaine and amphetamine regulated transcript peptide(CART),Ghrelin Receptor(GHSR)protein expression levels in hypothalamus by Western blotting(WB).Realtime fluorescence quantitative polymerase chain reaction(RTFQ-PCR)was used to detect the genes expression of NPY,AgRP,POMC,CART,OX1R,Ob-R and GHSR in hypothalamus.The expression levels of substance P(SP)and 5-hydroxytryptamine(5-HT)protein in duodenum were detected by Immunohistochemical technique.Results:After chemotherapy,the food intake of tumor-bearing mice decreased.After CHB-?-F treatment,the food intake,body weight and pathological changes of gastrointestinal mucosa were significantly improved compared with 5-FU group and negative control group.CHB-?-F combined with 5-FU could enhance the inhibitory effect of 5-FU on tumor.Compared with the tumor status of the negative control group,the tumor volume and the density of tumor cells in the other groups decreased to a certain extent.Amon g them,5-FU group and CHB-?-F(H)group had the most obvious inhibitory effect on tumor.The tumor inhibition rates of 5-FU group,YZXJC group,CHB-?-F(H),CHB-?-F(M)and CHB-?-F(L)group vwere 58.88%,28.08%,54.96%,37.69%and 28.61%,respectively.Compared with 5-FU group and negative control group,all the CHB-?-F-treated groups were higher than that of 5-FU group and negative control group.The contents of NPY,OrexinA.Ghrelin,GAS,MTL,EGF and PGE2 in serum of all the CHB-?-F-treated groups were higher than those of 5-FU group and negative control group.The content of Leptin in serum also decreased to some extent.Compared with the negative control group and 5-FU group,the activity of SOD in jejunum of all the CHB-?-F-treated groups were significantly increased,while the content of MDA in the jejunum was obviously lower than that in the negative control group and 5-FU control group.The results of Western blotting and RTFQ-PCR showed that compared with negative control group and 5-FU group,all the CHB-II-F-treated groups were higher than that of negative control group and 5-FU group,and the expression of OX1R,GHSR,NPY and AgRP proteins and genes in hypothalamus of all the CHB-?-F-treated groups were significantly higher than those of negative control group and 5-FU group.The protein expression and gene expression of Ob-R,POMC,CART in all the CHB-II-F-treated groups were significantly decreased,and the gene expression was basically consistent with the protein expression.Immunohistochemistry showed that the expression of 5-HT and SP protein in duodenum of mice with high,middle and low dose of CHB-II-F decreased significantly.Conclusion:CHB-II-F could not only enhance the inhibitory effect of 5-FU on liver cancer transplanted tumor mice,but also improve the pathological damage of Gastrointestinal mucosa induced by chemotherapy,and alleviate the attack of oxygen free radical on gastrointestinal tract.CHB-II-F could regulate the secretion of hormones in the gastrointestinal tract and relieve the side effects of chemotherapy on gastrointestinal toxicity caused by chemotherapy for tumor through affecting appetite regulators in the hypothalamus central nervous system feeding areas and peripheral appetite regulation factors. |
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