1.Review on Tanshinone Improving Intestinal Barrier Dysfunction in Sepsis
Journal of Zhejiang Chinese Medical University 2017;41(2):171-174
[Objective] To further study tanshinone role in intestinal barrier dysfunction in sepsis. [Methods] Mainly refer to nearly a decade at home and abroad on tanshinone pharmacological effects, and tanshinone in sepsis, especially the impact on the intestinal barrier dysfunction. [Results] Tanshinone in sepsis, and heart head blood-vessel, anti-tumor and so on in the field of study is more, but improving sepsis of intestinal barrier function in aspects of the study is less, but through tanshinone antibacterial anti-inflammatory, antioxidant, anti platelet aggregation, improve microcirculation, regulate immune function, this paper expounds tanshinone improving sepsis, possible mechanisms of intestinal barrier function. [Conclusion] Tanshinone can improve sepsis pathophysiological process, ischemia oxygen to the tissues and organs and whole body strong inflammatory reaction factors such as the cause of intestinal barrier function is impaired, but for its specific mechanism, further research is needed.
2.An observation on interference mechanism of Shenfu injection on ghrelin in rats with severe sepsis
Wan WU ; Ronglin JIANG ; Kan OUYANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2015;(6):596-600
Objective To observe the effect of Shenfu injection on intestinal function in rats with sepsis. Methods Forty Sprague-Dawley (SD) rats were randomly divided into four groups: sham operation, sepsis model, low and high concentration Shenfu injection groups, each groupn = 10. The sepsis model was replicated by cecal ligation and puncture (CLP), while the rate in sham operation group just underwent abdominal incision without CLP. Ten minutes after CLP, the low and high dose Shenfu injection groups were given 5 mL/kg and 10 mL/kg Shenfu intravenous injection via a tail vein respectively. The rats in the model group were treated by intravenous injection of 10 mL/kg normal saline through a tail vein in 10 minutes after CLP. Twelve hours later, the rats were sacrificed. The levels of Ghrelin, Gastrin, tumor necrosis factor-α (TNF-α), high mobility group B1 protein (HMGB1), myeloperoxidase (MPO) and diamine oxidase (DAO) activity in serum were detected by enzyme linked immunosorbent assay (ELISA). The levels of protein of Ghrelin and gastrin receptor (GHSR) were detected by Western Blot. Under light microscope, the histopathological changes in intestinal mucosa were investigated, and Chiu score was determined, and the apoptosis index (AI) of intestinal mucosal epithelial cells was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL).Results Compared with sham operation group, in model group, the levels of Ghrelin and Gastrin in serum were significantly decreased [Ghrelin (ng/L): 121.23±3.53 vs. 146.28±5.43, Gastrin (ng/L): 81.78±3.27 vs 102.78±4.07], the serum levels of TNF-α and HMGB1 were markedly increased [TNF-α (mg/L): 93.71±3.66 vs. 11.69±1.44, HMGB1 (mg/L): 76.25±4.12 vs. 22.41±3.08], the DAO activity and protein expressions of Ghrelin and GHSR of intestinal tissue were obviously decreased [DAO (U/mL): 14.64 ±0.68 vs. 25.13±1.98, Ghrelin (grey value): 0.12±0.02 vs. 0.23±0.04, GHSR (grey value): 0.18±0.02 vs. 0.32±0.03], the MPO activity in intestinal tissue, Chiu score of intestinal mucosa and AI of ileum mucosal epithelial cells were remarkably increased [MPO (mg/L): 175.98±6.95 vs. 45.64±4.48, Chiu score: 3.90±0.52 vs. 0.30±0.30, AI: 29.31±1.65 vs. 5.45±1.35, allP < 0.01]. Compared with model group, in low and high Shenfu injection groups, the levels of Ghrelin in serum and protein expressions of Ghrelin and GHSR in intestinal tissues were significantly increased (P < 0.05 orP < 0.01), the activity of DAO of intestinal tissues, the Chiu score and AI were significantly decreased, the degrees of changes being more significant in high Shenfu injection group than those in low Shenfu injection group [Ghrelin (ng/L): 143.54±3.89 vs. 136.58±4.91, TNF-α (mg/L): 75.13±4.69 vs. 83.70±4.40, HMGB1 (mg/L): 57.47±4.53 vs. 65.41±4.63, protein expression of Ghrelin (grey value): 0.18±0.03 vs. 0.15±0.03, protein expression of GHSR (grey value): 0.28±0.03 vs. 0.23±0.03, MPO (mg/L): 154.05±5.75 vs. 162.64 ±5.73, DAO (kU/L): 19.70±1.51 vs. 16.67±0.92, Chiu score: 2.30±0.52 vs. 3.20±0.48, AI: 20.38±1.34 vs. 26.40±1.32, allP < 0.05]. The levels of serum Gastrin in low and high Shenfu injection group were higher than those in model group, but no statistically significant differences were found (83.59±3.24, 86.54±5.93 vs. 81.78±3.27, bothP > 0.05). Under light microscope, the pathological changes were seen as follows: destruction and obvious edema of intestinal mucosal villi, ulcer formation, significant perivascular hemorrhage, presence of neutrophil infiltration and fracture of basement membrane in model group, while in low and high Shenfu groups, the intestinal villi had little defect, focal necrosis, small amounts of hemorrhage and neutrophil infiltration. Conclusions Shenfu injection can significantly improve the abnormal expressions of serum Ghrelin, reduce the levels of serum TNF-α and HMGB1, lowered MPO activity and enhance DAO activity in intestinal tissue, alleviate pathological changes in ileum mucosa, and decrease AI of ileum mucosal epithelial cells in rats with sepsis. And the degree of therapeutic effect is proportional to the Shenfu injection dose.
3.Protective Effect and Potential Mechanism of Astragalus Injection on Mucosa Epithelial Cells in Rats with Sepsis
Shuifang JIN ; Ronglin JIANG ; Shu LEI
Journal of Zhejiang Chinese Medical University 2015;(6):482-486
Objective] To investigate the protective effect and potential mechanism of Astragalus injection on mucosa epithelial cells in rats with sepsis . [Methods] Thirty Sprague-Dawley rats were randomly divided into sham,model and Astragalus injection treatment groups with 10 rats in each group.Rat sepsis model was constructed by cecal ligation and puncture.Rats in the Astragalus injection treatment group received 800 mg·kg-1 of Astragalus injection at 15 minutes before and 6 hours after operation. Intestinal mucosa were harvested and frozen at -80℃ on post-operation 24 hours. Intestinal mucosa epithelial cell apoptosis was detected by TUNEL,while Bcl-2 and Bax levels were determined by real-time quantitative PCR(qPCR) and Western blot technique.[Results] The apoptosis indexes were 5.2±0.3,22.3±1.2 and 11.2±0.9 of sham,model and Astragalus injection treatment group,respectively. Apoptosis index in model group was significantly higher than that in sham group( P<0.05),while the apoptosis of mucosa epithelial cells was significantly attenuated after Astragalus injection treatment(P<0.05).In model group,the expression of Bcl-2 was lower than that of sham group,whereas the expression of Bax was higher than that in sham group(P<0.05).Astragalus injection treatment significantly lowered Bax level while elevated Bcl-2 expression, compared with model group(P<0.05). [Conclusion] Astragalus injection could suppress sepsis-induced apoptosis of mucosa epithelial cells via up-regulating Bcl-2 and down-regulating Bax.
4.The intensive insulin therapy and different enteral nutrition in critically ill patients
Liquan HUANG ; Lincong WANG ; Ronglin JIANG
Parenteral & Enteral Nutrition 1997;0(04):-
0.05).The amount of insulin in ZL group was significantly lower than that in the control group(P
5.Effects of pneumonia caused by klebsiella pneumoniae on apoptosis of thymocytes in rats
Shu LEI ; Meifei ZHU ; Ronglin JIANG ; Jiannong WU
Chinese Journal of Emergency Medicine 2011;20(6):627-630
Objective To study the effects of pneumonia caused by Klebsiella pneumoniae on apoptosis of thymocytes in rats and its possible mechanism. Methods A total of 48 Sprague Dawley rats were randomly (random number) divided into 2 groups, namely the control group (n =24) and the infection group ( n = 24). The pneumonia models of rats were made with 0.3 mL Klebsiella pneumoniae suspension administered intratracheally per animal. On the 2nd, 4th, and 6th day after intratracheal instillation of bacteria, 1/ 3 of the rats in each group were sacrificed and TdT-mediated dUTP nick end labeling (TUN EL) method was used to assess the apoptosis of thymocytes. The expressions of cleaved Caspase-3, Bcl-2 and Fas in thympcytes of rats were detected with immunohistochemical staining. Results On each interval, apoptosis index of thymocytes, and the expressions of Cleaved Caspase-3 and Fas in the infection group were all higher than those in the control group (P < 0.01) , while the expressions of Bcl-2 lower than those in the control group (P<0.01). As more time consumed, the apoptotic index of thymocytes and the expressions of cleaved Caspase-3 in the infection group increased significantly (P<0.05). The expressions of Bcl-2 declined gradually (P < 0.05), but the expressions of Fas reached their peak 4th day after infection. There were no significant dynamic changes in all above mentioned variables in control group. Conclusions Pneumonia caused by Klebsiella Pneumoniae can lead to the increase in thymocyte apoptosis in rats. The mechanism may be associated with the decreased expression of Bcl-2 and the increased expression of Fas in thymocytes of rats with pneumonia caused by Klebsiella pneumoniae. The different apoptosis regulation pathways have different effects on different phase of pneumonia, that the effects of Fas decrease 4th day after pneumonia, while the effects of Bcl-2 increase further.
6.Anti-IL-5 monoclony antibody inhibits the migration of eosinophil in a murine model of asthma
Ronglin JIANG ; Huahao SHEN ; Shaobing WANG ; Pinli WANG ; Weibin MA
Basic & Clinical Medicine 2006;0(05):-
Objective To study the effect of anti-IL-5 monoclony antibody (TRFK-5) on migration of Eos from BM to the airways in sensitized mice. Methods Male C57BL/6 (6-8wk of age) murine model of Asthma and control group were estabolished with routine method. The outcome measurements include white blood cell (WBC) total count, differential count of bronchoalveolar lavage fluid (BALF) and peripheral blood (PB), nuclear cell count and eosinophil percentage of BM. These parameters were collected 12 h after the final allergen challenge. To cheek Eos infiltration, the histology of lung tissues was also observed. Further, the effects of intranasal TRFK-5 on above changes were investigated. Results Eosinophil numbers of BALF, PB, BM and the infiltration of Eos in pulmovnary tissues were increased considerably 12 h after final OVA challenge compared with negative group(P
7.Consensus of early enteral nutrition clinical practice in critically ill patients.
Renhua SUN ; Ronglin JIANG ; Man HUANG ; Guolong CAI
Chinese Critical Care Medicine 2018;30(8):715-721
The benefits of early enteral nutrition (EEN) during critical illness have been widely accepted by global experts. To popularize this new concept and provide standardized, reasonable and effective EEN therapy for critically ill patients in China, more than 20 experts from throughout the country discussed and developed this consensus. We used the GRADE approach for consensus development, focusing on important clinical issues such as nutrition assessment, initiating mode, route selection and tolerance monitoring of EEN support therapy for current critically ill patients. This consensus would be certainly help for intensive care physicians in the clinical application of EEN support therapy for critically ill patients.
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Enteral Nutrition
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8.Histone deacetylase inhibition induced epithelial-mesenchymal transition by Snail in hepatic oval cells
Qing HUANG ; Ronglin HU ; Xingyuan JIAO ; Heping PENG ; Xiaofeng JIANG ; Jun DU
Chinese Journal of Hepatobiliary Surgery 2012;(12):926-932
Objective Epithelial mesenchymal transition (EMT) has a role in the proliferation and metastasis of various types of cells.This study investigates the hepatic oval cell's mechanism of EMT induced by histone deacetylase inhibition and the resulting cell motility increase from EMT.Methods Hepatic oval cell stem cell markers and marker changes were detected by flow cytometry,and after histone deacetylase inhibition induced EMT,the morphological changes were recorded.Western blot and quantitative real-time polymerase chain reaction detected the expression of E-cadherin,vimentin and Snail.Furthermore,confocal microscopy analysis recognized the nuclear translocation of Snail.Results Flow cytometry revealed no changes in the stem cell properties of hepatic oval cells in the cell culture process.Oval cell EMT,induced by HDACi,was observed through morphological changes,a reduction of the epithelial cell marker E cadherin,and an increase of the mesenchymal cell marker Vimentin.HDACi can promote the expression and nuclear translocation of Snail,which is the key hepatic oval cell transcription factor for both EMT and enhanced motility.Therefore,Snail RNA interference can suppress HDACi induced EMT in hepatic oval cells.Conclusions In conclusion,histone deacetylase inhibition induces hepatic oval cell epithelial-mesenchymal transition by Snail.
9.Effect of moxibustion on N-methyl-D-aspartate receptor subtype 2B expression in hippocampus of rheumatoid arthritis model rats
Chuanyu PENG ; Ling HU ; Zijian WU ; Ronglin CAI ; Zhiming JIANG ; Yanping YANG
Journal of Acupuncture and Tuina Science 2022;20(3):174-180
Objective: To observe the effect of moxibustion on the expression of N-methyl-D-aspartic acid (NMDA) receptor subtype 2B (NR2B) in the hippocampus of rheumatoid arthritis (RA) rats, and to explore the analgesic mechanisms of moxibustion in RA treatment. Methods: Sixty male Sprague-Dawley rats were randomly divided into a normal group, a model group, a moxibustion group, a moxibustion + NMDA receptor antagonist (AP-5) group, and a moxibustion + NMDA receptor agonist (NMDA) group, with 12 rats in each group. Except for the normal group, rats in the other four groups were treated with complete Freund's adjuvant in a windy, cold, and damp environment to replicate RA models. Rats in the moxibustion group received suspended moxibustion with moxa sticks at Shenshu (BL23) and Zusanli (ST36), and the two points were used alternately. After intraperitoneal injection of AP-5 or NMDA, rats in the moxibustion + AP-5 group and the moxibustion + NMDA group received the same moxibustion intervention as in the moxibustion group, once a day for 15 d. The thermal withdrawal latency (TWL) of rats in each group was detected before and after modeling and after the 15-day intervention. After the 15-day intervention, hematoxylin-eosin staining was performed to observe the pathological changes in knee joints. The real-time fluorescence quantitative polymerase chain reaction method was used to detect the mRNA expression of NR2B in the hippocampus; Western blotting assay was used to detect the protein and the phosphorylated protein expression of hippocampal NR2B. Results: The synovial tissue was proliferated, the synovial lining was significantly thickened, the pannus was formed, and the cartilage and bone tissues were significantly damaged in the model group. After intervention, the pathological morphology of the knee joints in the moxibustion group, the moxibustion + AP-5 group, and the moxibustion + NMDA group was significantly improved, and the improvement in the moxibustion + AP-5 group was more notable than that in the moxibustion + NMDA group. Compared with the normal group, the TWL was significantly decreased (P<0.01), and the mRNA, protein, and phosphorylated protein expression levels of hippocampal NR2B were significantly increased in the model group (P<0.01). Compared with the model group, the TWL of each intervention group was significantly increased (P<0.01 or P<0.05), and the mRNA, protein, and phosphorylated protein expression levels of hippocampal NR2B were significantly decreased (P<0.01). Compared with the moxibustion group, the TWL was significantly increased (P<0.01), and the mRNA, protein, and phosphorylated protein expression levels of hippocampal NR2B were significantly decreased in the moxibustion + AP-5 group (P<0.01); the TWL was significantly decreased (P<0.01), and the mRNA, protein, and phosphorylated protein expression levels of hippocampal NR2B were significantly increased in the moxibustion + NMDA group (P<0.01). Conclusion: Moxibustion reduces hyperalgesia in RA inflammatory rats. The analgesic effect may be related to the decrease in the expression and phosphorylation levels of NR2B in the hippocampus.
10.Effect of Tanshinone ⅡA injection on intestinal mucosal tight junction protein in severe rat septic models
Wan WU ; Liquan HUANG ; Meifei ZHU ; Yihui ZHI ; Lingcong WANG ; Shu LEI ; Ronglin JIANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2018;25(1):93-98
Objective To discuss the influence of Tanshinone ⅡA on the tight junction protein of intestinal mucosal epithelial cells in rat severe septic models. Methods Seventy-five Sprague-Dawley (SD) rats were randomly divided into sham operation group, model group and Tanshinone ⅡA injection high (20 mg/kg), medium (10 mg/kg) and low (5 mg/kg) dose groups, each group 15 rats. Sepsis rat models were established by cecal ligation and puncture (CLP) method, in sham operation group, only switched abdominal surgery was performed without CLP. In Tanshinone ⅡA injection groups, different doses of Tanshinone ⅡA were injected intraperitoneally after modeling for 10 minutes and 6 hours; in sham operation and model groups, equal volume of normal saline was injected intraperitoneally at the same times as above. After operation, 3 L/kg of normal saline was injected into the caudal vein in all rats for fluid resuscitation.Twelve hours after operation, the rats were killed, the abdominal lymph nodes, liver, spleen and kidney tissues were taken for bacterial culture and calculating the rate of bacterial translocation; under microscope, the histopathological changes of ileum mucosal tissues were examined and Chiu scoring was carried out; TdT-mediated dUTP nick end labeling (TUNEL) was applied to detect the ileum mucosal epithelial cell apoptosis and calculating the index (AI);fluorescence immunoassay and Western Blot methods were used to measure the contents and protein expression levels of tight junction protein, junctional adhesion molecule-1 (JAM), Claudin-1, Zonula occludens-1 (ZO-1), Occludin, c-Fos and Tryptase. Results ① In bacterial cultures of abdominal lymph node, liver, spleen and kidney, the positive rate of mesenteric lymph node was the highest, followed by liver and spleen, mainly Escherichia coli, Proteus mirabilis, etc. The highest positive rate of bacterial culture was in model group (38.8%), followed by low dose of Tanshinone ⅡA injection group (35.0%), and the lowest was 16.6% in high dose Tanshinone ⅡA injection group, the differences being statistically significant in comparisons between any pair of groups (all P < 0.05). ② Pathological examination showed that the pathological changes of ileum mucosa were obvious and the Chiu score (4.17±0.98 vs. 0) and AI (11.70±2.87 vs. 2.17±0.80) in model group were significantly higher than those in sham group (all P < 0.05); with the increase of dosage of Tanshinone ⅡA injection, the pathological changes of rat ileum mucosa were improved gradually, the Chiu score and AI were decreased gradually, and the degrees of decrease in high dose Tanshinone ⅡA group were more significant than those in model group (Chiu score: 1.12±0.79 vs. 4.17±0.98, AI: 3.65±1.98 vs. 11.70±2.87, both P < 0.05).③ Immunofluorescence staining showed that the positive staining of protein JAM, ZO-1 and c-Fos were all green in color, Claudin-1, Occludin and Tryptase were all red in color, the localizations of all of them were in the cytoplasm, the protein expression of JAM, Claudin-1, ZO-1, Occludin from strong to weak in turn were Sham group, high, medium, low dose Tanshinone ⅡA group and model group, the expression of c-Fos, Tryptase from strong to weak in turn were model group, low, medium, high dose Tanshinone ⅡA group and Sham group. ④ Western Blot showed that the expressions of ileum tissue JAM, Claudin-1, ZO-1 and Occludin in model group were all significantly lower than those of the sham group, while the expressions of c-Fos, Tryptase were obviously higher than those of the sham group, with the increase of dosage of Tanshinone ⅡA, the expressions of JAM, Claudin-1, ZO-1 and Occludin were increased gradually and the protein expressions of c-Fos and Tryptase were gradually decreased, and the changes in high dosage group of Tanshinone ⅡA were more significant than those in low and moderate groups [JAM (gray value): 25.39±1.82 vs. 12.41±1.34, 19.45±1.66, Claudin-1 (gray value): 28.44±1.56 vs.17.26±1.46, 21.23±1.34, ZO-1 (gray value): 28.84±1.59 vs. 16.45±1.21, 24.22±1.46, Occludin (gray value): 25.49±1.63 vs. 13.34±1.45, 19.45±1.37, c-Fos (gray value):15.76±1.36 vs. 27.84±1.36, 21.22±1.73, Tryptase (gray value): 14.44±1.41 vs. 28.14±1.38, 22.32±1.57], all the above comparisons of different dosage groups were statistically significant (all P < 0.05). Conclusion Tanshinone ⅡA injection may improve intestinal wall structure and reduce bacterial translocation by improving the intestinal mucosal tight junction protein in sepsis model rats, and this effect is positively correlated to Tanshinone ⅡA dosage.