Protective Effect Of Hongteng Zhenqi Fuzheng Granule On Jejunum Injury In 5-FU Chemotherapy Colorectal Cancer Mice

Objective:(HPLC)was used to determine the controllability of HT185 drug quality.The in vivo experiment was conducted to study the effect of HT185 on 5-fluorouracil(5-FU)chemotherapy in colorectal cancer mice induced jejunum injury protection study for HT185 further drug development,clinical application to provide experimental basis.Methods:The HPLC fingerprints of HT185 were constructed by HPLC analysis,and the fingerprints of HT185 were determined.The chromatographic fingerprint similarity evaluation system was used to calculate the similarity of HPLC samples of different batches of HT185,and the quality of HT185 was evaluatedIn vivo animal experiments were performed on murine colon cancer CT-26 cells in logarithmic growth phase.Single cell suspension was prepared by trypan blue staining.The number of cells was counted at 2 × 106/mL and After subcutaneous inoculation,CT-26 cells were inoculated into the right anterior axillary part of BABL/c mice to establish an animal model of subcutaneous transplantation of colorectal cancer.The mice were randomly divided into 4 groups according to the size of the tumor volume,including the control group,HT185 group,5-FU group,5-FU + HT185,and the tumor volume was calculated to be 100-150mm3.Group,each group of 10.And treated with 5-FU(100 mg/kg)and HT185(12.4 g/kg),respectively.Among them,5-FU were given intraperitoneal injection once every other day after the day of grouping,and HT185 was given 1 day/day for 18 days on the day of grouping.Control group and HT185 group were given intragastric and intraperitoneal injection volume of saline.The weight of each group of mice was weighed every day and recorded well.The diarrhea of each group was observed daily and the scores were compared according to the diarrhea score index.On the 19th day,the mice were sacrificed by cervical dislocation,the jejunum was separated,fixed with 4%paraformaldehyde,embedded in paraffin,and cut with 4μm thickness.Then,the effect of HT185 on pathological changes of jejunum was observed by hematoxylin-eosin staining.The pathological changes of jejunum were sorted according to Macpherson and Pfeiffer histopathological scoring criteria.TUNEL method was used to detect HT185 On the apoptosis of intestinal cells induced by 5-FU.The effect of HT185 on the expression of Bcl-2 and Bax in jejunum crypts was detected by immunohistochemical staining(IHC).Results:1.The fingerprints of HT185 were established,and the HPLC fingerprints of different batches of HT185 were similar,the drug composition was stable and the quality of HT185 was controlled.2.Compared with the control group and HT185 drug group,the weight of 5-FU mice decreased significantly,and most of the diarrhea symptoms appeared on the third day after injection of 5-FU;the paraffin histopathological sections could be observed(P<0.05).There was no significant difference in the expression of intestinal mucosal lesions,such as intestinal villi arrangement irregularities,rupture,inflammatory cell infiltration,intestinal epithelial cells,lamina propria,edema,(P<0.05).The expression of Bax was significantly increased(P<0.05).Compared with the 5-FU group,the expression of Bcl-2 was significantly lower than that of the 5-FU group(P<0.05),the degree of mucosal injury in jejunum was reduced,and the number of apoptotic cells in the jejunal glands was significantly lower than that in the control group(P<0.05),but the effect of FU + HT185 on the weight loss of the rats was significantly improved(P>0.05).The expression of Bcl-2 protein was significantly increased and the expression of Bax protein was significantly decreased(P<0.05).3.There was no significant difference between the control group and the control group(P>0.05).There was no significant difference between the control group and the control group(P>0.05).Conclusion:1.Establishment of HT185 fingerprints,HPLC analysis of different batches of HT185,after similarity calculation,HT185 quality control.2.The effect of HT185 on 5-FU-induced jejunum injury can significantly improve the diarrhea of mice and reduce the degree of mucosal injury in jejunum.3.HT185 can reduce the apoptosis of jejunum crypts and regulate the expression of Bcl-2 and Bax,which are related to mitochondrial apoptosis pathway,and thus protect 5-FU-induced jejunum injury,which may be the effect of HT185 on 5-FU induced jejunum One of the protective mechanisms of injury.