| Purpose:To study whether saikosaponin-d can inhibit IL-6,IL-1?and PGE2-related inflammatory factors and reduce cisplatin-induced inflammatory damage of gastric mucosa in rats;To investigate whether saikosaponin-d influences PI3k/AKT signaling pathway.Regulates BCL-2 protein and protects the gastric mucosa.Material and method:Thirty-six 160-180g SD rats were selected and provided by Liaoning Experimental Animal Resource Center?Liaoning Changsheng Biotechnology Co.,Ltd.?and randomly divided into three groups:blank group,cisplatin group,saikosaponin d+cisplatin In the group?Chinese medicine group?,10 rats in each group were housed in separate cages,and 3 groups of experiments were performed at the same time.This experiment was based on the literature[1]dose method for the administration of cisplatin 6mg/kg tail vein injection,saikosaponin d 6.5 mg/kg intragastric administration(this experiment according to the literature[2,3]choose 3.5mg The drug concentrations were compared at 5kg/kg,6.5mg/kg,and 6.5mg/kg was determined as the optimal drug concentration.The blank group was given intragastric gavage,Cisplatin group was given 1-4days of cisplatin injection,5-10 days of normal saline was given by gavage,Chinese medicine group was given 1-3 days of saikosaponin d gavage,and 4-7 days were given cisplatin plus bupleurum Saponin d,8-10 days continue to give saikosaponin d,co-administration 10 days,24h after fasting water was killed,leaving the stomach specimens.According to the above subgroups,gavage was performed once a day?ig?,and ig was given regularly and continuously,and the rats were free to drink water during the experimental period.Each day,the weights of rats before and after administration were weighed and recorded.The weight gain and loss values of each rat before and after administration were calculated,and the gastric mucosal damage of the rats was observed.The rat gastric mucosal injury index was calculated,and HE staining and immunization were performed.IHC,ELISA and Western-Blot were used to detect the expression of related inflammatory factors and their proteins in gastric mucosal injury,and to explore the possible mechanism of saikosaponin d to protect gastric mucosa.Statistical Analysis:SPSS19.0 software was used for statistical processing.Chi-square test was used for count data analysis.Measured data were expressed as meanąstandard deviation and t-test was used.P<0.05 was considered statistically significant.Results:1.Rats'own condition:In the blank group,the rats were normal in spirit,normal in activity,and had no other symptoms;the rats in the cisplatin group were apathetic,reduced in activity,hairless and sparse,loose stools,irritability,irritability,and cramps;The spirit of the Chinese medicine group rats can not significantly reduce the activity,the hair was no sparse shedding and shiny,loose stool thin reduced significantly.2.Body weight:The body weight of the blank group increased;body weight of the cisplatin group was significantly reduced;weight loss of the Chinese medicine group was improved.3.The gastric mucosal injury index?UI?:After dissection,the blank group was observed with naked eyes.No flatulence or enlargement was observed in the stomach.The mucosal surface of the stomach tissue showed a pale red color,the mucosal surface was smooth and complete,and there was no congestion,edema and damage,and no bleeding points;In the platinum group,gastric enlargement was observed,there was a large amount of food residue,there was obvious inflation,gastric mucosal damage was slightly heavier,mucosal color was red,local lesions,redness,and there was a small amount of bleeding spots on the mucosal surface,but less,the degree of visual damage was not very serious The injury index score was higher than that of the blank group;the stomach of the Chinese medicine group had no food residue and no obvious flatulence.Compared with the same period,the gastric tissue lesions of the cisplatin group were relieved,the mucosal damage was lighter,the mucous membrane was pale pink,intact and not damaged.There was no significant spotting.The injury index score was lower than that of the cisplatin group.4.HE:In the blank group,the glandular structure of the gastric mucosa was intact,the cells were arranged neatly,the capillaries were abundant,the layers were clear and continuous,and no inflammatory cell infiltration was observed;the cisplatin group was damaged in the gastric mucosal gland structure,and most of the cells were shedding.Disorderly arrangement,blood vessel rupture and visible infiltration of a large number of red blood cells,submucosal inflammatory cell infiltration;Chinese medicine group gastric mucosal glands are absent or there is less shedding,the general glands are arranged neatly,there is no red blood cell infiltration,intracellular congestion has improved The infiltration of inflammatory cells was significantly reduced and there were significant differences in statistics?P<0.05?.5.Immunohistochemistry?IHC?:Compared with the blank group,the contents of PI3k and AKT proteins in the cisplatin group and the traditional Chinese medicine group were significantly increased,and the Chinese medicine group was higher than the cisplatin group,and the Bcl-2 protein content in the cisplatin group and the Chinese medicine group was decreased.However,the Chinese medicine group was higher than the cisplatin group,and the difference was statistically significant?P<0.05?.6.Western-Blot:Compared with the blank group,the p-PI3k and p-AKT protein levels in the cisplatin group and the traditional Chinese medicine group were significantly increased,and the Chinese medicine group was higher than the cisplatin group,and the Bcl-2 protein content was both in the cisplatin group and the Chinese medicine group.Lower than the blank group,but the Chinese medicine group was higher than the cisplatin group,the difference was statistically significant?P<0.05?.7.ELISA:The expression of IL-6 and IL-I?in cisplatin group was higher than that in blank group and Chinese herb group.PGE2 level was lower than that in blank group and Chinese medicine group,the difference was statistically significant?P<0.05?.Conclusion:1.Traditional Chinese medicine Bupleurum extract saikosaponin d can increase the level of PGE2 after gastric mucosal injury,decrease the expression of IL-6 and IL-I?,and reduce the inflammatory damage of gastric mucosa;2.Saikosaponin d may affect the expression of Bcl-2 protein,reduce the damage and protect the gastric mucosa by affecting PI3K/AKT signaling pathway. |
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