| Objective Severe acute pancreatitis(SAP)has always been a worldwide problem due to the dangerous condition,complex pathogenesis,high mortality and plenty complications.Qingyi decoction has been clinically verified in the treatment of SAP through multiple practices,but the specific mechanism is not clear yet.Hyperbaric oxygen therapy has been widely used in the treatment of some acute and chronic diseases,but it has not been widely and deeply studied in the treatment of SAP.The purpose of this study was to observe the effects of hyperbaric oxygen,Qingyi decoction and the combination of hyperbaric oxygen with Qingyi decoction on SAP in rats by establishing the model of SAP in rats and explore the mechanism of the different therapies.Hyperbaric oxygen treatment combined with other routine treatment was used to explore the clinical effects and future applications of hyperbaric oxygen for patients with moderate SAP(MSAP).Methods Healthy male Wistar rats of clean grade were randomly divided into 5 groups:Normal control group(N group),severe acute pancreatitis model group(SAP group),Qingyi decoction treatment group(Z group),hyperbaric oxygen treatment group(H group)and the combination of hyperbaric oxygen with Qingyi decoction treatment group(HZ group).Rats in SAP group,Z group,H group and HZ group were all induced SAP by retrograde injection of 3.5%sodium taurocholate into pancreaticobiliary duct.The first treatment time of traditional Chinese medicine(Qingyi decoction)in group Z and HZ group was 30 minutes(once a day,1ml/100g)after 3 days of abdominal closure.The first time of hyperbaric oxygen therapy in group H and HZ group was 6 hours after abdominal closure.The time of each treatment was 100 minutes and the pressure in hyperbaric oxygen animal chamber was 2 ATM,once a day for 3 days.At the end of the experiment,the survival of rats in each group was first recorded.The pancreas,ileum,serum,ascites and feces of rats were retained.The levels of serum amylase and lipase in rats were measured by automatic biochemical analyzer.The levels of interleukin-6(IL-6),interleukin-8(IL-8)and tumor necrosis factor-α(TNF-α)in serum were measured by enzyme-linked immunosorbent assay.The pathological damage of pancreas and ileum was detected by hematoxylin-eosin(HE)staining and pathological score was performed and calculated.The expression of antibacterial peptide-lysozyme in ileal Pan’s cells was detected by real-time fluorescence quantitative RT-PCR.The abundance and diversity of intestinal microbial communities were analyzed by 16s rDNA V3-V4 sequencing.In the clinical study,hyperbaric oxygen treatment group and routine treatment control group were set up.There were 52 cases in the former group(47 MSAP and 5SAP);50 cases in the routine group(46 MSAP and 4 SAP).The patients in 2 groups were treated with inhibiting pancreatic enzyme secretion,correcting electrolyte disturbance,anti-inflammatory,traditional Chinese medicine,inferior nasogastric tube,abdominal puncture and drainage or pancreatic necrosis clearance.In addition to the above treatments,patients in the hyperbaric oxygen treatment group were treated with hyperbaric oxygen once a day for 120 minutes,7 times as a course.C-reactive protein was measured within 24 hours after admission and after the treatment of the3~rdd and 7~thh hyperbaric treatment in both groups.Evaluation of APACHE II score within 24 hours after admission,3~rdd and 7~thh hyperbaric oxygen therapy were taken;Balthazar CT grading score within 24 hours after admission and after every 7hyperbaric oxygen therapy were taken.Results 1.The mortality rates of N group,SAP group,Z group,H group and HZ group were0.00%,60.00%,53.33%,53.33%and 40.00%,respectively.The levels of ascites,serum amylase,serum lipase,IL-6,IL-8 and TNF-αin SAP group were significantly higher than those in N group(P<0 05).Compared with SAP group,the above indexes in H group,Z group and HZ group were significantly lower than those in SAP group(P<0 05).Furthermore,HZ group showed the most effective results.2.Pathological lesions in pancreas and ileum were observed in SAP group.Compared with SAP group,the pathological damage of pancreas and ileum in H group,Z group and HZ group were significantly improved,and the histopathological scores were significantly decreased(P<0.01).The improvement was the most obvious in HZ group(P<0.001).3.The expression of antibacterial peptide-lysozyme in ileal Pan’s cells in SAP group was significantly lower than that in N group(P<0.001),and the expression of antibacterial peptide-lysozyme in ileal Pan’s cells in HZ group,H group and Z group was higher than that in SAP group,among which HZ group was the highest and had statistical difference(P<0.001).The diversity and abundance of intestinal microbial communities in SAP group were significantly lower than those in N group,while those in HZ group,H group and Z group were higher than those in SAP group,among which HZ group was the closest to N group.4.The results of human treatment showed that the there was no significant difference in APACHEII scores between the hyperbaric oxygen treatment group and the routine treatment group at admission(P>0.05),but there was significant difference on the 3~rdd and 7~thh day(P<0.05).Significant difference in the level of C-reactive protein between the two groups appeared on the 7~thh day(P<0.05).No significant difference in Balthazar CT score was observed(P>0.05).Conclusions 1.The SAP rat model induced by retrograde injection of 3.5%sodium taurocholate into pancreaticobiliary duct was confirmd by pathological examination with pancreatic parenchyma necrosis,hemorrhage,edema and inflammatory cell infiltration in different degrees.The model was reproduceable and reliable.2.Hyperbaric oxygen therapy and Qingyi decoction treatment have effects on reducing mortality,improving the severity of the disease and maintaining the stability of intestinal microecology.The combination of hyperbaric oxygen with Qingyi decoction has the most significant effect.3.Clinical experiments showed that hyperbaric oxygen treatment group was effective in improving APACHEII score and reducing C-reactive protein level compared with the routine treatment.There was no effect in improving Balthazar CT grade. |
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