| Objective: The mechanism of CAG and the intervention of Xiangsha Liujunzi Decoction were explored from the perspective of PI3K/AKT signaling pathway key molecules by investigating the expression of PTEN,PI3 K,AKT,BAD,BCL-XL,BAX genes and proteins in gastric mucosa of CAG rats,so as to provide scientific basis for the clinical application of Xiangsha Liujunzi Decoction.Methods: 120 healthy Wistar rats,SPF grade,2 months old,weighing(200 ±20)g,half male and half female,were randomly divided into blank group(n = 20)and m odel group(n = 100).The CAG model was replicated by comprehensive method(MNNG + ranitidine + anorexia).The rats were randomly divided into model group(n = 16),vitacoenzyme tablets group(n = 16),high dose group of Xiangsha Liujunzi Decoction(n = 16),medium dose group of Xiangsha Liujunzi Decoction(n = 16),low dose group of Xiangsha Liujunzi Decoction(n = 16).The model group and the blank group were given 10 m L/(kg·d)distilled water by gastric lavage.The high,middle and low dose groups of Xiangsha Liujunzi Decoction were given 24,12,6 g/(kg·d)Xiangsha Liujunzi Decoction by gastric lavage,and the vitacoenzyme tablets group was given 0.03 g/(kg·d)vitamin preparations by gastric lavage.After 120 days of administration,the general survival status and pathological sections of gastric antrum mucosa of rats in each group were evaluated.Quantitative PCR was used to detect PTEN,PI3 K,AKT,BAD,BCL-XL and BAX gene expression in rat gastric mucosa.Western blot was used to detect the expression levels of PTEN,PI3 K,AKT,p-AKT,BAD,BCL-XL and BAX protein in rat gastric mucosa.Result: 1.General living conditions: The rats in the blank group were generally in good condition during the modeling stage.Rats in CAG model group generally had poor survival status,showing bow bending,squinting,emaciation,lethargy,anorexia,sluggishness,sparse hair,dull and dull coat,less or loose stool.At the stage of drug treatment,the rats in the blank group were generally in good condition and gradually enlarged in body shape;the symptoms of the high-dose group of Xiangsha Liujunzi Decoction improved significantly;the genera l survival condition of rats in medium dose group and vitacoenzyme tablets group of Xiangsha Liujunzi Decoction were partly improved;and the general survival of the rats in the low-dose group of Xiangsha Liujunzi Decoction did not improve significantly.2.Pathological changes: Compared with the blank group,the gastric mucosa of the model group became thinner,the epithelial cells and glands arranged disorderly and sparsely,the number of glands decreased significantly,the muscular layer of the mucosa became thicker and extended to the lamina propria,and a large number of inflammatory infiltration could be seen.Compared with the model group,the pathological morphology of the rats in the high-dose group,the medium-dose group and the vitacoenzyme tablets group improved in varying degrees,especially in the high-dose group of Xiangsha Liujunzi Decoction.The pathological morphology of rats in the low dose group of Xiangsha Liujunzi Decoction was not significantly improved.3.RT-PCR test results:(1)Compared with the blank group,the relative expression levels of PTEN and BAX genes in gastric mucosa of rats in model group were significantly lower(P < 0.01);compared with model group,the relative expression levels of PTEN and BAX genes in gastric mucosa of rats in high-dose Xiangsha Liujunzi Decoction group and medium-dose Xiangsha Liujunzi Decoction group were significantly higher,and the relative expression levels of BAX genes in gastric mucosa of rats in vitacoenzyme tablets group were significantly higher(P < 0.01).The relative expression levels of PTEN and BAX genes in gastric mucosa of rats in Shaliujunzi Decoction low dose group increased,while those in vitacoenzyme tablets group increased,but there was no significant difference between groups(P > 0.05).(2)Compared with the blank group,the relative expression levels of PI3 K,AKT,BAD and BCL-XL genes in gastric mucosa of model group rats were significantly increased(P < 0.01);compared with model group,the relative expression levels of PI3 K,AKT,BAD and BCL-XL genes in gastric mucosa of high-dose Xiangsha Liujunzi Decoction group rats were significantly decreased,while the relative expression levels of BAD and BCL-XL genes in gastric mucosa of middle-dose Xiangsha Liujunzi Decoction group rats and Weienzymatic group rats were significantly decreased(P < 0.01).The relative expression levels of PI3 K and AKT genes in gastric mucosa of rats in medium dose group,low dose group and vitacoenzyme tablets group decreased significantly(P < 0.01 or P < 0.05),while the relative expression levels of BAD and BCL-XL genes in gastric mucosa of rats in low dose group decreased significantly,but there was no significant difference between groups(P > 0.05).4.Western-blotting test results:(1)Compared with the blank group,the relative expression levels of PTEN and BAX protein in the gastric mucosa of the model group were significantly lower(P < 0.01 or P < 0.05);compared with the model group,the relative expression levels of PTEN and BAX protein in the gastric mucosa of the high-dose group of Xiangsha Liujunzi Decoction and the medium-dose group of Xiangsha Liujunzi Decoction were significantly higher,and the relative expression levels of BAX protein in the gastric mucosa of the vitacoenzyme tablets group were significantly higher(P < 0.0).The relative expression level of BAX protein in gastric mucosa of rats in low dose group of Xiangsha Liujunzi Decoction increased,while that of PTEN protein in gastric mucosa of rats in vitacoenzyme tablets group increased,but there was no significant difference between groups(P > 0.05).(2)Compared with the blank group,the relative expression levels of PI3 K,AKT,p-AKT,BAD and BCL-XL protein in the gastric mucosa of the model group were significantly increased(P<0.01);compared with the model group,the relative expression levels of PI3 K,AKT,p-AKT,BAD,BCL-XL protein in the gastric mucosa of the high-dose Xiangsha Liujunzi Decoction group were significantly decreased,while the relative expression levels of PI3 K,AKT,p-AKT,BCL-XL protein in the middle-dose Xiangsha Liujunzi Decoction group were significantly decreased.The relative expression levels of AKT,BAD and BCL-XL proteins in gastric mucosa of rats in the vitacoenzyme tablets group decreased significantly(P < 0.01 or P < 0.05);the relative expression levels of BAD proteins in gastric mucosa of rats in the medium dose group of Xiangsha Liujunzi Decoction decreased,and the relative expression levels of PI3 K,AKT,p-AKT,BAD and BCL-XL proteins in the low dose group of Xiangsha Liujunzi Decoction were lower.The relative expression levels of PI3 K and p-AKT protein in gastric mucosa of rats in the vitacoenzyme tablets group decreased,but there was no significant difference between groups(P > 0.05).Conclusion: 1.Xiangsha Liujunzi Decoction can improve the general survival of CAG rats.2.Xiangsha Liujunzi Decoction can improve the pathological state of gastric mucosa in CAG rats.3.Xiangsha Liujunzi Decoction can down-regulate the relative expression levels of AKT,PI3 K,BAD,BCL-XL genes and proteins in PI3K/AKT signaling pathway in gastric mucosa of CAG rats,and up-regulate the relative expression levels of PTEN,BAX genes and proteins in PI3K/AKT signaling pathway,which may play a therapeutic role by inhibiting cell proliferation and promoting apoptosis. |
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