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ObjectiveTo investigate the therapeutic effect of Qingjie Huagong decoction （QJHGD） on a mouse model of severe acute pancreatitis （SAP） and the mechanism of action of QJHGD against inflammatory response. MethodsA total of 36 male C57BL/6J mice were randomly divided into blank group， model group， Western medicine group （ulinastatin）， and low-， middle-， and high-dose QJHGD groups， with 6 mice in each group. All mice except those in the blank group were given 5% sodium taurocholate by retrograde pancreaticobiliary injection to establish a model of SAP. After modeling， the mice in the low-， middle-， and high-dose groups were given QJHGD （1， 2， and 4 g/kg， respectively） by gavage， and those in the Western medicine group were given intraperitoneal injection of ulinastatin （5×104 U/kg）， for 7 days in total. HE staining was used to observe the histopathological changes of the pancreas； ELISA was used to measure the levels of α-amylase， lipase， interleukin-1β （IL-1β）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， interleukin-18 （IL-18）， and tumor necrosis factor-α （TNF-α） in mice； RT-qPCR was used to measure the mRNA expression levels of NOD-like receptor protein3 （NLRP3）， Toll-like receptor 4 （TLR4）， and nuclear factor-kappa B （NF-κB） in pancreatic tissue； immunohistochemistry was used to measure the positive expression rates of NLRP3， TLR4， and NF-κB in pancreatic tissue； Western blot was used to measure the protein expression levels of NLRP3， TLR4， NF-κB， IL-1β， and IL-6. An analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsCompared with the blank group， the model group had diffuse destruction of pancreatic tissue structure， focal dilatation of pancreatic lobular septum， pancreatic acinar atrophy， and massive inflammatory cell infiltration， as well as significant increases in the content of α-amylase， lipase， IL-1β， IL-6， IL-8， IL-18， and TNF-α （all P<0.05）， the mRNA expression levels and positive expression rates of NLRP3， TLR4， and NF-κB （all P<0.05）， and the protein expression levels of NLRP3， TLR4， NF-κB， IL-1β， and IL-6 （all P<0.05）. Compared with the model group， the low-， middle-， and high-dose QJHGD groups and the Western medicine group had slightly tighter and more intact structure of pancreatic tissue， ordered arrangement of pancreatic acinar cells， a small amount of inflammatory cell infiltration， and hemorrhagic foci of pancreatic lobules， as well as significant reductions in the content of α-amylase， lipase， IL-1β， IL-6， IL-8， IL-18， and TNF-α （all P<0.05）， the mRNA expression levels and positive expression rates of NLRP3， TLR4， and NF-κB （all P<0.05）， and the protein expression levels of NLRP3， TLR4， NF-κB， IL-1β， and IL-6 （all P<0.05）. ConclusionQJHGD may exert a protective effect on the pancreatic tissue of SAP mice by inhibiting the activation of NLRP3/TLR4/NF-κB signaling pathway-related proteins， reducing the release of inflammatory mediators， and preventing the enhancement of inflammatory cascade response.
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Objective:To evaluate the clinical efficacy of TCM Qingjie Huagong Decoction combined with routine internal medicine in the treatment of severe acute pancreatitis with cholelithiasis (bile duct stones) in the early stage.Methods:Thirty-two patients with severe acute pancreatitis combined with cholelithiasis in the first affiliated Hospital of GuangXi University of Traditional Chinese Medicine were selected and randomly divided into two groups with 16 in each, both groups were treated for 14 days. Serum amylase (AMS) was detected by iodine-starch colorimetry, GOT and GPT were detected by continuous monitoring method, and CRP, IL-6 and procalcitonin (PCT) were detected by immune transmission turbidimetry. Acute Physiological and Chronic Health Score Ⅱ (APACHE Ⅱ), CT Severity Index Score (CTSI) and Modified Marshall Score were used to evaluate the severity of SAP. The recovery time of body temperature, the relief time of abdominal distension pain, the recovery time of bowel sounds and the total hospital stay were observed and recorded to evaluate the clinical effect.Results:The total effective rate was 93.8% (15/16) in the treatment group and 75.0% (12/16) in the control group. There was significant difference between the two groups ( χ2=8.19, P=0.042). After treatment, the level of AMS, WBC, CRP, PCT, AST, ALT and IL-6 in the treatment group were lower than those in the control group ( t values were 14.3, 7.24, 9.63, 5.48, 7.05, 7.33, 28.34, respectively, all Ps<0.05); After treatment, the time for body temperature to return to normal [(2.91±0.12)d vs. (3.78±0.38)d, t=8.76], the time for relief of abdominal distension pain [(4.77±0.68)d vs. (7.13±1.55)d, t=9.52], the time for recovery of bowel sounds [(3.90±1.80)d vs. (4.89±1.38)d, t=2.98] and the total hospital stay [(22.60±2.80)d vs. (30.37±3.89)d, t=7.88] in the treatment group were all significantly shorter than those in the control group ( P<0.01); APACHE Ⅱ, CTSI and the Modified Marshall Score in the treatment group were lower than those in the control group ( t values were 11.82, 12.72, 7.71, respectively, all Ps<0.01). Conclusion:Qingjie Huagong Decoction combined with ERCP and conventional western medicine therapy can reduce the level of inflammation in patients with cholelithiasis in the early stage of SAP, relieve clinical symptoms and improve clinical efficacy.
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OBJECTIVE To explore the the reg ulation of intestinal flora and effects of Qingjie huagong decoction on intestinal mucosal barrier in severe acute pancreatitis （SAP）mode rats . METHODS SAP rat model was induced by intraperitoneal injection of caerulein and lipopolysaccharide.The survival state of rats in each group were observed.The levels of serum amylase ，interleukin 10（IL-10），IL-18 and IL- 1β in serum were all detected. The pathological changes of pancreatic and small intestinal tissue were observed. The expressions of Occludin，ZO-1 and HMGB1 were detected in small intestinal tissue of rats. The structure and relative abundance of intestinal microflora in rats were detected by 16S rRNA high throughput sequencing. RESULTS After the intervention of Qingjie huagong decoction ，abdominal distension symptoms of SAP model rats were significantly relieved ，and their mental state recovered better ；the levels of serum amylase and IL- 18 in serum were decreased significantly （P＜0.05），while the level of IL- 10 was increased significantly （P＜0.05）. The necrotic area of pancreatic tissue and the infiltration of inflammatory cells were reduced ， the degree of intestinal epithelial cell structural disorder was alleviated ，and the shedding of intestinal mucosal epithelium was reduced.The protein expression of HMGB 1 in small intestinal tissue was decreased significantly （P＜0.05），and the protein expression of Occludin and ZO- 1 were increased significantly . Results of 16S rRNA high throughput sequencing showed that Qingjie huagong decoction could increased the relative abundance of probiotics such as Bacteroidea and Lactobacillus in rat intestine ，reduced the colonization of harmful bacteria such as Firmicutes. CONCLUSIONS Qingjie huagong decoction can improve the intestinal barrier by up-regulating the expression of Occludin and ZO- 1 in small intestinal tissue and down-regulating the protein expression of HMGB 1. It can also adjust the relative abundances of different flora to protect the intestinal tract.
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Background: Previous studies suggest that ghrelin plays an important role in the pathogenesis of acute pancreatitis, but the mechanism is still unclear. Aims: To screen the differentially expressed genes in pancreatitis pancreatic acinar cells transfected with ghrelin miRNA by gene chip. Methods: Pancreatitis was induced by caerulin. AR42J cells were divided into ghrelin miRNA+caerulin group and negative control+caerulin group. AR42J cells were collected to extract RNA and synthesize ds-DNA, and was then hybridized with rat whole genome microarray for scanning and analyzing. Differentially expressed genes related to inflammation and calcium pathways were screened, and expressions of Bcl-2, caspase-8, caspase-12 were verified by RT-PCR. Results: For AR42J cells after intervention with ghrelin miRNA+caerulin, 2 938 differentially expressed genes were screened, including 1 435 up-regulated genes and 1 503 down-regulated genes. There were 60 differentially expressed genes involved in CAMP/Ca2+ signaling pathway and 199 differentially expressed genes involved in inflammation pathway. RT-PCR results showed that expressions of Bcl-2, caspase-12 were down-regulated and expression of caspase-8 was up-regulated in ghrelin miRNA+caerulin group, consistent with the results of gene chip. Conclusions: Gene chip can be used to screen the genes that play a key role in the process of pancreatitis pancreatic acinar cells transfected with ghrelin miRNA, and provide a theoretical basis and reference for the study of mechanism of endogenous ghrelin on pancreatitis pancreatic acinar cells.
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Objective To investigate the roles of toll like receptor7 (TLR7) and toll like receptor 9 (TLR9) in the pathogenesis of acute pancreatitis.Methods AR42J cells were treated by lipopolysaccharide at different dosages (0,1,10,100 mg/L),and cell model of acute pancreatitis in vitro was established.AR42J cells without lipopolysaccharide treatment were as control.Cells and culture supernatant were collected after 24 hours cultivation.TLR7,TLR9 mRNA and protein expressions were detected by RT-PCR and Western Blot,and levels of TNF-α,IL-10 in culture supernatant were measured by ELISA.Results The TLR 7 mRNA expression levels in control group,1,10,100 mg/L lipopolysaccharide group were 0.12 ± 0.09,0.28 ± 0.06,0.49 ± 0.04,0.78 ± 0.04,and the TLR9 mRNA expression levels were 0.06 ± 0.02,0.32 ± 0.03,0.56 ± 0.14,0.84 ± 0.12; the TLR7 protein expression levels were 0.04 ± 0.01,0.26 ± 0.05,0.49 ±0.04,0.77 ±0.16,and the TLR9 protein expression levels were 0.10 ±0.14,0.62 ±0.23,1.21 ± 0.26,1.75 ± 0.13 ; the TNF-α levels in culture supernatant were (8.01 ± 5.32),(25.64 ± 8.71),(49.06 ± 10.23),(75.83 ± 6.65) ng/L,and the IL-10 levels were (155.54 ± 25.47),(105.16 ± 10.49),(69.36 ± 8.19),(14.07 ± 9.06)ng/L.The expression levels of TLR7 and TLR9's mRNA,protein in cell,as well as the levels of TNF-α in culture supernatant increased with the lipopolysaccharide concentration,while the levels of IL-10 in culture supernatant decreased with the lipopolysaccharide concentration,and the difference among these groups was statistically significant (P ＜ 0.01).Conclusions The expressions of TLR7 and TLR9 in AR42J cells treated by using lipolysaccharide are obviously up-regulated,and it suggests that TLR7 and TLR9 may be vital in the pathogenesis of acute pancreatitis.
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Objective To observe the expression of adrenomedullin (ADM) mRNA in hepatic tissue of rats with acute necrotizing pancreatitis (ANP) complicated with hepatic injury.Methods Sixty-four SD rats were randomly divided into control group and ANP group with 32 rats in each group.In ANP group,ANP model was induced by retrograde injection of 5％ sodium taurocholate into biliopancreatic duct of rats.Rats in control group only received sham operation and pancreas manipulation.All the rats were sacrificed at 3,6,12,24 h after the operation.The serum levels of amylase,alanine aminotransferase (ALT),aspartate aminotransferase (AST) and ADM were detected.Pathological changes in pancreatic and hepatic tissue were examined.The expressions of ADM mRNA in hepatic tissue were evaluated by fluorescence quantitative PCR.Results The serum concentrations of amylase,ALT,AST were (7229 ±968),(174.2 ±28.0),(657.7 ± 139.0) U/L,which were significantly higher than those in control group [(2036 ± 292),(104.3 ± 22.1),(419.7 ± 86.3) U/L],and the difference between the two groups was statistically significant (P ＜ 0.05 or P ＜0.01).Pathological injury of pancreas and liver tissue in ANP gradually aggravated with time,and the pathological scores at 12 h were (11.60 ± 1.51),(2.60 ± 0.89),which were significantly higher than those in control group (1.20 ± 0.77,0),and the difference between the two groups was statistically significant (P＜0.01).The serum concentrations of ADM in ANP group increased at 3 h after ANP induction,and reached (38.53 ± 6.25)pg/ml at 12 h,which was significantly higher than that in control group [(28.99 ±3.92)pg/ml] ; the concentrations of ADM in liver tissue increased at 3 h after ANP induction,and reached (3.00 ± 1.49) at 6 h,which was significantly higher than that in control group (1.04 ± 0.20),and the difference between the two groups was statistically significant (P＜0.05 or P＜0.01).Conclusions The expression of ADM mRNA in rat 's hepatic tissue increases in the early stage of ANP,and the serum concentration of ADM also increases.
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Objective To observe the expression of ghrelin and growth hormone secretagogue receptor (GHSR) in pancreas of rats with acute necrotizing pancreatitis (ANP),and investigate the role of ghrelin in the pathogenesis of ANP.Methods Seventy SD rats were randomly divided into ANP group (n =35) and control group (n =35).ANP model was induced by retrograde injection of 4％ sodium taurocholate into the biliary and pancreatic duct.Rats in the control group underwent laparotomy with gentle pancreas manipulation only.At 3,6,12,24,48 h after ANP induction,the rats were sacrificed,and the serum level of amylase was determined,pathological changes in pancreatic tissue were routinely observed and scored.The expressions of ghrelin mRNA,protein and GHSR mRNA,protein in pancreas were evaluated by RT-PCR and Western blot.Results Serum amylase level began to increase at 3h after sodium taurocholate injection and reached the peak value at 6 h [(8244 ± 2950) U/L],which was significantly higher than that in control group (P ＜ 0.05).Pancreatic injuries was aggravated with time,the pathologic score at 24 h was (11.91 ± 1.31) score,which was significantly higher than (3.12 ± 1.60) score in the control group.The expressions of ghrelin mRNA and GHSR mRNA in pancreas of ANP group were increased gradually with time,and were significantly higher than those of control group at all time points,at 48 h,1.29 ±0.64 vs 0.58 ±0.05 and 0.94 ±0.16vs 0.19 ±0.03,P ＜ 0.05.The expressions of ghrelin protein and GHSR protein in pancreas of ANP group were significantly higher than that in control group at 12,24,48 h.at 48 h,3.05 ± 0.48 vs 2.18 ± 0.23 and 2.34 ± 0.32 vs 1.55 ± 0.10 (P ＜ 0.05).Conclusions The expressions of ghrelin mRNA,protein and GHSR mRNA,protein of pancreas are significantly increased in rats of ANP,and are associated with the severity of ANP.
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Objective To investigate the prevalence and risk factors of chronic kidney disease (CKD) in the adult urban population of Hezhou Guangxi. Methods One thousand and two hundred urban residents (older than 18 years) from Hezhou Guangxi were randomly selected using a random sampling. All the residents were interviewed. Their morning spot urine were tested to determine albumin to ereatinine ratio (abnormal:≥30 mg/g), and renal function [abnomal: eMDRD <60 ml·min-1·(1.73 m2)-1] was assessed. Morning spot urine dipstick of hematuria (abnormal:≥1 +) was confirmed by microscopy (abnormal: 3 red blood cells/HP). The associations among demographic characteristics, health eharacteristies and indicators of kidney damage were examined. Results Eligible data of 1069 subjects were enrolled in the study. The prevalence of albuminuria was 7.5%, hematuria 4.8%, and reduced eGFR 3.6%. The prevalence of kidney disease was 14.4% and the recognition was 1.4%. Age (OR 1.022, 95%CI 1.008-1.035), gender (OR 2.249, 95%CI 1.502-3.367), diabetes mellitus (OR 7.422, 95%CI 3.985-13.825) and hypertension (OR 4.397, 95% CI 2.601-7.432) were independently associated with CKD. Conclusions The prevalence of chronic kidney disease is 14.4% and the recognition is 1.4% in adult urban population of Hezhou Guangxi. Independent risk factors associated with chronic kidney disease are age, gender, diabetes mellitus and hypertension which is similar to those in developed countries and domestic big cities.