The Mechanism Of Compound Qizao Deco-Ction In The Therapy Of Acetic Acid-induced Gastric Ulcer Of Rats

ObjectiveIn this study,we want to detect the therapeutic effect of Compound qizao decoction in acetic acid-induced gastric ulcer of rats;and discuss the portection mechanism of gastric mucosa by detecting the expressions of oxidative stress related factors MDA,SOD and GSH-PX,as well as the expressions of apoptosis-related factors Fox03a and bim.Methods1.Modeling:60 rats were randomly divided into normal control group,ulcer group,FFQZT low dose group(3 g/kg),FFQZT medium dose group(6 g/kg),FFQZT high dose group(12 g/kg)and Omeprazole group(3.6 mg/kg).Preparation of ulcer model was established by acetic acid cauterization.Rats in each group were anesthetized with 1%pentobarbital sodium.After anesthesia,rats were fixed in supine position,the upper abdominal hair was cut off with curved scissors,and the surgical area was exposed and disinfected with iodine.The abdomen was cut 1.5 cm below the xiphoid process,and a longitudinal incision needed to be done to expose the muscular layer.The shears were cut along the left side of the midline of the abdomen by 0.5 cm.The abdominal cavity was probed with a toothless forceps,and the stomach was gently pulled out of the body after being found behind the liver.A piece of filter paper was dipped in glacial acetic acid and attached to the junction of the antrum body of the anterior wall of the stomach for 30 s.The blood vessels should be avoided during the attachment.After removal,the new filter paper was dipped in glacial acetic acid and attached to the same site for 30 s.the attached part was washed with normal saline,and the washing fluid should be avoided into the abdominal cavity.After rinsing residual glacial acetic acid,it was dried with sterile gauze and the adherent site was covered with omentum majus.Then the tissues were sutured layer by layer and disinfected with iodide again.On the third day,the administration groups were administered the above-mentioned dose of 1 mL/100 g by gavage for 14 days,and the normal group and the ulcer group were given the same dose of saline(0.9%NaCl).2.Sampling:After the last administration,the rats were fasted for 12 hours and anesthetized with 2 mL/100 g of sodium pentobarbital.Rats were fixed in the supine position after anesthesia.Blood was taken from the abdominal aorta and centrifuged at 3000 rpm for 10 min.The supernatant was collected,stored at-80 ?.The stomach of the rats were cut along the large curvature of the stomach,and the stomach cavity was rinsed with iced saline,and then flattened on an ice bag,and the morphology of the stomach wall was observed and photographed.The maximum diameter and width of ulcer lesion were measured with vernier calipers.The ulcers were divided into two parts,which were separately stored in 4%paraformaldehyde or at-80?.3.Detection indicators:TBA method was used to determine the content of MDA in serum.The activity of SOD in serum was determined by WST-1 method.Serum GSH-PX activity was determined by colorimetry.The expressions of Fox03a and Bim were determined by immunohistochemistry and Western blotting.TUNEL assay was used to determine the apoptosis rate of gastric mucosal epithelial cells.Results1.General view of gastric tissue:The gastric mucosa of rats in the normal group was smooth,and the mucosal folds were in natural motion,without hyperemia or swelling,showing pink color.The gastric mucosa of rats in the model group showed round ulcer-like lesions with uplifts around and depressions in the center,with clear boundaries,white moss covering the center of the lesions and a slightly hard tactile sensation,and the mucosa color was slightly paler than that of the normal group.With the application of drugs,the area of ulcer foci decreased significantly,the degree of hyperemia and edema was weakened,,the depth of depression was reduced,and the plica of eminence were gradually flattened.In the FFQZT group and the omeprazole group,the ulcer foci almost disappeared,and the ulcers affected the wrinkles and returned to normal.2.Ulcer index and ulcer inhibition rate:Compared with the model group,the UI of FFQZT groups and omeprazole group was significantly reduced(P<0.01).Compared with the omeprazole group,the FFQZT high-dose group had a higher ulcer inhibition rate(P<0.01),while the difference in the low dose group was not statistically significant(P>0.05).3.Oxidative stress indexes:Compared with the normal group,MDA content was increased,the SOD activity was decreased and GSH-PX activity was decreased in the model group(P<0.01).Compared with the model group,MDA content in the middle and high dose groups of FFQZT decreased(P<0.01),GSH-PX activity increased,SOD activity increased(P<0.05).Compared with omeprazole group,FFQZT medium and high dose group GSH-PX activity increased,the difference was statistically significant(P<0.05),and there was no significant difference in MDA content and SOD activity in FFQZT groups(P>0.05).4.Immunohistochemical stainings:Fox03a was mainly expressed in the nucleus,while Bim was mainly expressed in the cytoplasm.In the normal group,the positive expressions of Fox03a and Bim were low and the staining degrees were light.The positive expressions of Fox03a and Bim were significantly increased and the staining degrees were deeper in the ulcer group.The expressions of Fox03a and Bim in FFQZT group and omeprazole group decreased to different degrees.Compared with the ulcer model group,the expressions of Fox03a and Bim were significantly reduced in the FFQZT high-dose group(P<0.01).5.Western blotting:Compared with the normal group,FoxO3a and Bim protein expressions were significantly increased in the model group(P<0.01).Compared with the model group,the protein expressions of Fox03a in FFQZT group and omeprazole group were significantly reduced(P<0.01),the expression of Bim in FFQZT high-dose group was significantly reduced(P<0.01),the expression of Bim between FFQZT low-dose and medium-dose group were not statistically significant(P>0.05).6.TUNEL results:Compared with the normal group,the apoptosis rate in the model group was significantly increased(P<0.01).Compared with the model group,the apoptosis rates of FFQZT group and omeprazole group were reduced to different degrees(P<0.01).Compared with the omeprazole group,the apoptosis rate of the FFQZT high-dose group was significantly reduced(P<0.01),while there was no significant difference of the apoptosis rate in the FFQZT medium-dose group(P>0.05).Conclusions1.Compound qizao decoction has a good therapeutic effect on acetic acid-induced gastric ulcer in rats.2.Oxidative stress is involved in the formation of gastric ulcer.The compound qizao decoction could exert its anti-ulcer effect by reducing MDA content,increasing SOD and GSH-PX activity,and resisting oxidative stress.3.Apoptosis is involved in the formation of gastric ulcer.The compound qizao decoction can inhibit the apoptosis of gastric mucosal epithelial cells by regulating the Fox03a?Bim protein expression,thereby exerting its anti-ulcer effect.