Qingyi decoction attenuates intestinal epithelial cell injury via the calcineurin/nuclear factor of activated T-cells pathway.

BACKGROUND: Recent studies have demonstrated that dysfunction of the intestinal barrier is a significant contributing factor to the development of severe acute pancreatitis (SAP). A stable intestinal mucosa barrier functions as a major anatomic and functional barrier, owing to the balance between intestinal epithelial cell (IEC) proliferation and apoptosis. There is some evidence that calcium overload may trigger IEC apoptosis and that calcineurin (CaN)/nuclear factor of activated T-cells (NFAT) signaling might play an important role in calcium-mediated apoptosis. AIM: To investigate the potential mechanisms underlying the therapeutic effect of Qingyi decoction (QYD) in SAP. METHODS: A rat model of SAP was created via retrograde infusion of sodium deoxycholate. Serum levels of amylase, tumor necrosis factor (TNF-α), interleukin (IL)-6, D-lactic acid, and diamine oxidase (DAO); histological changes; and apoptosis of IECs were examined in rats with or without QYD treatment. The expression of the two subunits of CaN and NFAT in intestinal tissue was measured via quantitative real-time polymerase chain reaction and western blotting. For in vitro studies, Caco-2 cells were treated with lipopolysaccharide (LPS) and QYD serum, and then cell viability and intracellular calcium levels were detected. RESULTS: Retrograde infusion of sodium deoxycholate increased the severity of pancreatic and intestinal pathology and the levels of serum amylase, TNF-α, and IL-6. Both the indicators of intestinal mucosa damage (D-lactic acid and DAO) and the levels of IEC apoptosis were elevated in the SAP group. QYD treatment reduced the serum levels of amylase, TNF-α, IL-6, D-lactic acid, and DAO and attenuated the histological findings. IEC apoptosis associated with SAP was ameliorated under QYD treatment. In addition, the protein expression levels of the two subunits of CaN were remarkably elevated in the SAP group, and the NFATc3 gene was significantly upregulated at both the transcript and protein levels in the SAP group compared with the control group. QYD significantly restrained CaN and NFATc3 gene expression in the intestine, which was upregulated in the SAP group. Furthermore, QYD serum significantly decreased the LPS-induced elevation in intracellular free Ca2+ levels and inhibited cell death. CONCLUSION: QYD can exert protective effects against intestinal mucosa damage caused by SAP and the protective effects are mediated, at least partially, by restraining IEC apoptosis via the CaN/NFATc3 pathway.

Qingyi decoction protects against myocardial injuries induced by severe acute pancreatitis.

BACKGROUND: We studied the protective effects of Qingyi decoction (QYD) (a Traditional Chinese Medicine) against severe acute pancreatitis (SAP)-induced myocardial infarction (MI). AIM: To study the function and mechanism of QYD in the treatment of myocardial injuries induced by SAP. METHODS: Ultrasonic cardiography, hematoxylin and eosin staining, immunohistochemistry, qRT-PCR, western blot, enzyme-linked immunosorbent assays, and apoptosis staining techniques were used to determine the effects of QYD following SAP-induced MI in Sprague-Dawley rats. RESULTS: Our SAP model showed severe myocardial histological abnormalities and marked differences in the symptoms, mortality rate, and ultrasonic cardiography outputs among the different groups compared to the control. The expression of serum cytokines [interleukin (IL)-1ß, IL-6, IL-8, IL-12, amyloid ß, and tumor necrosis factor-α] were significantly higher in the SAP versus QYD treated group (P < 0.05 for all). STIM1 and Orai1 expression in myocardial tissue extracts were significantly decreased post QYD gavage (P < 0.001). There was no significant histological difference between the 2-aminoethyl diphenylborinate inhibitor and QYD groups. The SAP group had a significantly higher apoptosis index score compared to the QYD group (P < 0.001). CONCLUSION: QYD conferred cardio-protection against SAP-induced MI by regulating myocardial-associated protein expression (STIM1 and Orai1).

Yi Guan Jian decoction may enhance hepatic differentiation of bone marrow­derived mesenchymal stem cells via SDF­1 in vitro.

A previous study reported that Yi Guan Jian (YGJ) may increase the proliferation and differentiation of hepatic oval cells in a rat liver cirrhosis model. The aim of the present study was to investigate the effect and mechanism of action of YGJ on inducing hepatic differentiation in bone marrow­derived mesenchymal stem cells (BM­MSCs) via stromal­cell derived factor­1 (SDF­1). Murine BM­MSCs were isolated with whole bone marrow adherence, then identified by immunocytochemical staining and flow cytometry. Passage 2 cells were divided into 8 groups and their differentiation was induced by cell factors added to the medium, including hepatocyte growth factor (HGF), SDF­1 and YGJ. Each of the cell factors was used alone and any two or three of them were combined to establish different cell microenvironments in the different treatment groups. Albumin (ALB) was selected as a hepatocellular marker and cytokeratin­18 (CK­18) as a cholangiocellular marker. The protein and mRNA expression levels of ALB and CK­18 were used to determine the differentiation of BM­MSCs using immunocytochemical staining, western blotting and reverse transcription­quantitative polymerase chain reaction on days 7, 14, 21 and 28 during induction. The relative expression levels of ALB and CK­18 resulted in time­dependent increases in the groups supplemented only with HGF, SDF­1 or YGJ. Combination treatment of any two HGF, SDF­1 and YGJ led to a higher expression of ALB and CK­18 compared with only one cell factor treatment. Additionally, when all three were used in a combined treatment the expression levels of ALB and CK­18 occurred at an earlier time and was higher overall. Therefore, the present study suggested that YGJ had an effect on inducing hepatic differentiation in BM­MSCs via SDF­1 and may act in a synergistic manner with HGF and SDF­1.

Effect of Yi Guan Jian decoction on differentiation of bone marrow mesenchymalstem cells into hepatocyte-like cells in dimethylnitrosamine-induced liver cirrhosis in mice.

Yi Guan Jian decoction (YGD) may induce the differentiation of bone marrow mesenchymal stem cells (BMSCs) into hepatocyte-like cells (HLCs); however, the underlying mechanisms remain to be elucidated. The present study aimed to investigate this process. To do this, a dimethylnitrosamine (DMN)-induced liver cirrhosis mouse model was established. The mice from the model group were randomly divided into three subgroups: i) Negative control, ii) hepatocyte growth factor and iii) YGD. The overall health, liver function and histological alterations were monitored. The expression of α­smooth muscle actin (α­SMA), C­X­C chemokine receptor type 4 (CXCR4), extracellular signal­regulated kinase (ERK1/2), nuclear factor κB p65 subunit (NF­κB p65) and ß­catenin were measured by immunohistochemistry, western blotting and reverse transcription­quantitative polymerase chain reaction. Following administration of DMN, the overall health of the mice significantly decreased, with an increase in pathological developments and liver damage resulting in a decrease in liver function. Immunohistochemistry revealed that the expression of α­SMA, CXCR4, ERK1/2, NF­κB p65 and ß­catenin was upregulated. Following treatment with YGD, the overall health, liver function and pathology improved. The mRNA and protein expression levels of CXCR4 and ERK1/2 were upregulated, where as α­SMA, NF­κB p65 and ß­catenin levels were downregulated. The results demonstrated that YGD may induce the differentiation of BMSCs into HLCs to reverse DMN­induced liver cirrhosis; this may be achieved via an upregulation of the SDF­1/CXCR4 axis to activate the mitogen activated protein kinase/ERK1/2 signaling pathway.

[Distributions of phosphorus fractions in suspended sediments and surface sediments of Tiaoxi mainstreams and cause analysis].

Phosphorus is a primary nutrient showing the water quality status of river and inducing eutrophication, and a different phosphorus fraction can make diverse contributions to water quality. Four phosphorus forms of suspended sediments and surface sediments in Tiaoxi mainstreams were measured using a sequential extraction procedure, and the distributions of their forms were discussed. The results showed that the tropic status of Xitiaoxi River was inferior to that of Dongtiaoxi River as a whole, and the water quality in the middle reach of Dongtiaoxi River was better than that in the upper and lower reaches. The contents of nutrient elements in suspended sediments were significantly higher than those in surface sediment, which indicated an enrichment of nutrient in fine sediment. The percentages of the loosely absorbed phosphorus ( NH4Cl-P), the reductant phosphorus (BD-P) and the metal oxide bound phosphorus (NaOH-P) in the suspended sediment were higher than those in surface sediment, while the percentage of the calcium bound phosphorus (HCl-P) showed a reverse trend. Correlation analyses between phosphorus forms and chemical compositions of suspended sediments and surface sediments were performed. The results showed the phosphorus forms in suspended sediments and surface sediments of Xitiaoxi River had weak relationships with mineral components, while those in the Dongtiaoxi River had strong relationships with mineral, especially OM and clay mineral. The cause was associated with the geological setting and material sources in Tiaoxi watershed.

Therapeutic effect of Qingyi decoction in severe acute pancreatitis-induced intestinal barrier injury.

AIM: To investigate the effect of Qingyi decoction on the expression of secreted phospholipase A2 (sPLA2) in intestinal barrier injury. METHODS: Fifty healthy Sprague-Dawley rats were randomly divided into control, severe acute pancreatitis (SAP), Qingyi decoction-treated (QYT), dexamethasone-treated (DEX), and verapamil-treated (VER) groups. The SAP model was induced by retrograde infusion of 1.5% sodium deoxycholate into the biliopancreatic duct of the rats. All rats were sacrificed 24 h post-SAP induction. Arterial blood, intestine, and pancreas from each rat were harvested for investigations. The levels of serum amylase (AMY) and diamine oxidase (DAO) were determined using biochemical methods, and serum tumor necrosis factor (TNF)-α level was measured by an enzyme linked immunosorbent assay. Pathologic changes in the harvested tissues were investigated by microscopic examination of hematoxylin and eosin-stained tissue sections. The expressions of sPLA2 at mRNA and protein levels were detected by reverse transcriptase PCR and Western blot, respectively. A terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay was used to investigate apoptosis of epithelial cells in the intestinal tissues. RESULTS: Compared to the control group, the expression of sPLA2 at both the mRNA and protein levels increased significantly in the SAP group (0.36 ± 0.13 vs 0.90 ± 0.38, and 0.16 ± 0.05 vs 0.64 ± 0.05, respectively; Ps < 0.01). The levels of AMY, TNF-α and DAO in serum were also significantly increased (917 ± 62 U/L vs 6870 ± 810 U/L, 59.7 ± 14.3 ng/L vs 180.5 ± 20.1 ng/L, and 10.37 ± 2.44 U/L vs 37.89 ± 5.86 U/L, respectively; Ps < 0.01). The apoptosis index of intestinal epithelial cells also differed significantly between the SAP and control rats (0.05 ± 0.02 vs 0.26 ± 0.06; P < 0.01). The serum levels of DAO and TNF-α, and the intestinal apoptosis index significantly correlated with sPLA2 expression in the intestine (r = 0.895, 0.893 and 0.926, respectively; Ps < 0.05). The levels of sPLA2, AMY, TNF-α, and DAO in the QYT, VER, and DEX groups were all decreased compared with the SAP group, but not the control group. Qingyi decoction intervention, however, gave the most therapeutic effect against intestinal barrier damage, although the onset of its therapeutic effect was slower. CONCLUSION: Qingyi decoction ameliorates acute pancreatitis-induced intestinal barrier injury by inhibiting the overexpression of intestinal sPLA2. This mechanism may be similar to that of verapamil.

[Expression of Cav-1, AQP1 and AQP5 in lung of acute pancreatitis-associated lung injury rats and the therapeutic role of Qingyitang].

OBJECTIVE: to investigate the expression and location of caveolin-1 (Cav-1), aquaporin 1 (AQP1) and AQP5 in the lung of acute pancreatitis-associated lung injury rats and to determine the role of these molecules in the pathologic progress and the potential therapeutic mechanisms of Qingyitang. METHODS: forty Wistar rats were randomly divided into sham operation (SHAM) group, acute lung injury (ALI) group, dexamethasone (DEX) group and Qingyitang (QYT) group. ALI was induced by reverse injection of deoxycholate into biliopancreatic duct of rats. Blood and lung tissues were collected after 24 h. Serum amylase, lung wet/dry (W/D) ratio and pathological section were detected to evaluate the degree of lung injury. reverse transcription-polymerase chain reaction was taken to detect the mRNA levels of Cav-1, AQP1 and AQP5. Lipid rafts were prepared for detection of the distribution and expression level of Cav-1, AQP1 and AQP5 proteins by Western blot. RESULTS: the concentration of serum amylase, the value of W/D and the degree of pathological lung injury obviously increased in ALI rats. Cav-1, AQP1 and AQP5 were present in the lung while the mRNA level decreased in ALI rats. Cav-1 appeared mainly in lipid rafts and less in non-lipid rafts. AQP1 was localized to lipid rafts while AQP5 to non-lipid rafts. The localization of these three molecules in the lung of ALI rats did not change compared with SHAM rats while their protein levels decreased. Compared with ALI rats, the concentration of serum amylase, the value of W/D and the degree of pathological lung injury obviously decreased in DEX and QYT rats. The mRNA and the protein expression of Cav-1, AQP1 and AQP5 increased in various degrees by DEX or QYT treatment. CONCLUSION: Cav-1 and AQP1 are enriched in lipid rafts while AQP5 in non-lipid rafts. The down-regulated expression of these three molecules may play important role in acute pancreatitis-associated lung injury. DEX and QYT may relieve lung injury effectively by up-regulating the expressions of Cav-1, AQP1 and AQP5.

[Expression of secretory type II phospholipase A2 in acute lung injury following acute pancreatitis and interventional effect of Qingyi decoction on it].

OBJECTIVE: To investigate the expression of secretory type II phospholipase A(2) (sPLA(2)-II) in lung of rats with acute lung injury (ALI) complicating severe acute pancreatitis (SAP), and the effect of Qingyi decoction (QYT) on ALI. METHODS: Thirty Sprague Dawley (SD) rats were randomly divided into three groups: sham operation (SO) group, model group and QYT group, with 10 rats in each group. SAP model was reproduced by reverse injection of sodium deoxycholate into the common bile- pancreatic duct of rats. The pancreas of rats was just exposed in SO group. QYT (10 ml/kg) was gavaged 30 minutes and 12 hours after SAP was induced in QYT group. The blood gas analysis was performed 24 hours after operation. Serum amylase (AMY) levels, sPLA(2) and lung wet/dry ratio (W/D) were determined. The sPLA(2)-II mRNA and sPLA(2)-II protein expression in lung were detected by reverse transcription- polymerase chain reaction (RT-PCR) and Western blotting. The pathological changes in lung and pancreas were observed. RESULTS: Compared with SO group, the levels of arterial partial pressure of oxygen (PaO(2)) and pH value in model group were significantly decreased [PaO(2) (mm Hg, 1 mm Hg=0.133 kPa): 79.24±5.84 vs. 96.78±3.81, pH value: 7.269±0.054 vs. 7.391±0.054], arterial partial pressure of carbon dioxide (PaCO(2)), the serum levels of AMY, W/D ratio and the serum levels of sPLA(2) were significantly increased [PaCO(2) (mm Hg): 47.57±2.55 vs. 27.69±1.02, AMY (U/L): 7 144.19±727.91 vs. 1 193.41±192.54, W/D ratio: 8.57±2.45 vs. 3.70±0.90, sPLA(2) (nmol×min(-1) ×ml(-1)): 45.13±6.05 vs. 29.94±6.39], the expression of sPLA(2)-II mRNA (1.28±0.21 vs. 0.80±0.08) and protein were significantly increased (all P <0.05). Compared with model group, blood PaO(2) and pH value were significantly increased [PaO(2): (88.16±5.07) mm Hg, pH value: 7.322±0.039], the PaCO(2), the serum levels of AMY, W/D ratio and the serum levels of sPLA(2) in QYT group were significantly decreased [PaCO(2): (33.13±2.14) mm Hg, AMY: (4 283.51±527.52) U/L, W/D ratio: 4.05±0.52, sPLA(2): (28.00±4.78) nmol×min(-1) ×ml(-1)], and the expression of sPLA(2)-II mRNA (0.89±0.08) and protein were significantly decreased (all P <0.05). The pathological changes in lung and pancreas in QYT group were milder than those in SAP group. CONCLUSION: The higher expression of sPLA(2)-IIin lung may be one of pathogenetic factors in ALI induced by SAP. Administration of QYT can reduce the injury of lung by decreasing the expression of sPLA(2)-II in transcriptional level and thus protecting pulmonary function.

Effect of combined therapy of Yinchenhao Chengqi decoction and endoscopic sphincterotomy for endotoxemia in acute cholangitis.

AIM:To evaluate the therapeutic mechanism of Yinchenhao Chengqi (YCHCQ) decoction (containing mainly Herba Artemisia capillaris) combined with endoscopic sphincterotomy (EST) for endotoxemia (ETM) in acute cholangitis.METHODS:Twenty-one cases of acute cholangitis with ETM were divided randomly into two groups: group A, 10 patients treated with YCHCQ decoction combined with EST, group B,11 patients treated with EST. The incidence rate of ETM, plasmic ET, serum superoxide dismutase (SOD) activity, malonyldialdehyde (MDA), complement C(3) and C-reactive protein (CRP) were studied respectively.RESULTS:The ET level of group A (35.92ng/L ± 8.30ng/L) was significantly reduced after 7 days of treatment (P < 0.05) in contrast to that of group B (47.8ng/L ± 11.62ng/L), so did the level of MDA and CRP. But the SOD activity and C(3) level in group A increased significantly (P < 0.05).CONCLUSION:YCHCQ decoction combined with EST had a beneficial effect for ETM in acute cholangitis.