Effects of mesenteric lymph duct ligation on sepsis-induced lung and intestinal injuries in rats / 西安交通大学学报(医学版)

Objective To study the effect of mesenteric lymph duct ligation(MLDL)on sepsis-induced lung and intestinal injuries in rats.Methods Sepsis was induced by cecal ligation and puncture(CLP)in male rats.The rats were randomly divided into sham control group in which rats received sham operation,sepsis group(CLP group)in which rats received saline after CLP operation,and CLP+MLDL group in which rats received mesenteric lymph duct ligation(MLDL)after CLP operation.The rats were sacrificed at 24 h after CLP modeling. Bronchoalveolar lavage fluid,arterial blood,and intestine and lung tissues were collected to determine organ injuries.Results The lung tissue histopathology and blood gas analysis revealed that MLDL markedly alleviated the lung injury(ALI score:6.14 ± 0.51 vs.8.12 ± 0.63,P=0.029 9),improved oxygen partial pressure[(78.67 ± 4.51)mmHg vs.(64.83 ± 2.90)mmHg,P=0.0273],decreased lung W/D ratio(6.12 ± 0.25 vs.7.63 ± 0.49,P=0.021 2),decreased BALF cytokines levels[TNF-α:(828.17 ± 81.89)pg/mL vs.(1 118.17 ± 79.22)pg/mL,P=0.029 1;IL-6:(39.33 ± 5.50)ng/mL vs.(68.67 ± 5.24)ng/mL,P=0.003 4],alveolar permeability[protein levels:2.117 ± 0.289 2 vs.3.033 ± 0.164 7,P=0.020 3,and cell levels:(30.00 ± 3.587)×106/L vs.(43.83 ± 2.358)×106/L,P=0.009 1].However,according to the results of HE and biochemical detection,MLDL had no protective effect on sepsis-induced intestinal injury.Moreover,MLDL could significantly reduce the bacterial loads of the blood and lung,but do not change the bacterial level in the intestines.Conclusion MLDL has a significant protective effect on sepsis-induced lung injury mainly by decreasing bacterial translocation,but had no effect on intestinal injury.This difference may be related to that MLDL inhibits the bacteria spreading from abdominal cavity to other organs in sepsis but it causes their accumulation in the intestines.

Effect of ligation of mesenteric lymph duct on coagulation function in rats with severe heat stroke / 解放军医学杂志

Objective To investigate the influences of mesenteric lymph duct ligation (LDL) on the coagulation function in rats with severe heat stroke (SHS). Methods Forty male Wistar rats were randomly and equally divided into control, heat stroke sham (HSS), HSS-LDL, severe heat stroke (SHS) and SHS+LDL groups. Mesenteric lymph ducts were ligated in HSC-LDL and SHS-LDL groups before reproduction of SHS model. SHS rat models were reproduced in a prewarmed incubator. Peripheral blood was drawn to determine the parameters pertaining to blood coagulability including prothrombin time (PT), activated partial thromboplastin time (APTT), D-dimmer and platelet count (PLT), and lung and kidney histopathology was observed after heat stroke. Results Compared with those in control group, no obvious changes were observed in the coagulation indices in HSS and HSS-LDL group. While PT and APTT significantly prolonged, PLT remarkably decreased, D-dimmer markedly increased in SHS group (P<0.05). The coagulation indices presented a recovery trend to certain extent in SHS-LDL group. Histopathological examination of the kidney and lung showed severe hemorrhagic and congestive lesions in SHS group. Mesenteric lymph duct ligation alleviated the coagulation disorders and histopathological lesions. Conclusion The entrance of toxic agents through lymphatic passage may be the pathogenetic factor in producing coagulopathy, and ligation of the mesenteric lymph ducts might prevent the entrance of toxic materials and alleviate the injury to the blood coagulation property and internal organs.

Effect of mesenteric lymphatic ligation on liver injury induced by severe acute pancreatitis in rats / 实用医学杂志

Objective To explore the effect of mesenteric lymphatic ligation on liver injury induced by severe acute pancreatitis. Methods 54 SD rats are randomly divided into sham-operation group (A), severe acute pancreatitis group (B) and severe acute pancreatitis and ligation group (C). Liver tissues were collected from each rat in 6, 12, and 24 h after ligation. Pathological changes in liver tissues were observed by haematoxylin and eosin staining. Levels of serum glutamic-pyruvic transaminase and glutamic oxalacetic transaminase were detected. Levels of TNF-α and IL-1 in liver homogenate were examined by enzyme linked immunosorbent assay. Results Under light microscopic examination, neutrophils and inflammatory exudate in hepatic lobule was increased as time prolonged in group B. Inflammation was reduced in group C as compared with group B.Levels of serum glutamic-pyruvic transaminase and glutamic oxalacetic transaminase were more significantly increased in groups B and C than in group A (P<0.05);while they were more significantly decreased in group C than in group B. Levels of TNF-αand IL-1 in liver homogenate were significantly increased in group B as compared with group A (P < 0.05); whereas they were more significantly decreased in group C than in group B at 24 h (P < 0.05). Conclusions Mesenteric lymphatic ligation can reduce liver injury in severe acute pancreatitis. Mesenteric lymphatic fluids may play an important role in severe acute pancreatitis.

A study on alleviation of acute lung injury in rats with bowel repletion pattern by mesenteric lymph drainage and its mechanism / 中国中西医结合急救杂志

Objective To observe the effects of mesenteric lymph drainage on acute lung injury and expression of p38 mitogen activated protein kinase (p38MAPK) signal pathway in rats with bowel repletion pattern. Methods Thirty male Wistar rats were randomly divided into three groups according to random number table method, namely sham operation group (sham group), bowel repletion model group (model group) and mesenteric lymph drainage group (drainage group), 10 rats in each group. The rat model of bowel repletion was established by ischemia/reperfusion (I/R) method, firstly 1 hour occlusion of superior mesenteric artery (SMA) to induce ischemia followed by reperfusion for 2 hours. In the rats of drainage group, the drainage of mesenteric lymph duct began at the end of model establishment and persisted for 3 hours. In the rats of sham group, the SMA and mesenteric lymph ducts were exposed with blunt dissection, and then they were immediately placed back into the abdominal cavity. After 3 hours of mesenteric lymph drainage, the lung and ileum tissues of rats in each group were harvested for evaluation of pathohistological changes and for the determination and comparison of myeloperoxidase (MPO) activity changes; the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in bronchoalveolar lavage fluid (BALF) were detected by enzyme linked immunosorbent assay (ELISA), and the expressions of Toll-like receptor 4 (TLR4) mRNA and p38MAPK mRNA in the lung tissues were measured by fluorescence quantitative polymerase chain reaction (PCR).Results Under the light microscope, the pulmonary capillaries markedly dilated and congested, the interstitium width of lung increased with a large amount of inflammatory cells infiltration, the intestinal mucosal layer becoming thinner with detachment of intestinal villi and a large amount of inflammatory cells infiltration were detected in rats of model group. Compared with those in sham group, the levels of TNF-α and IL-6 in BALF, the MPO activity of lung and ileum tissues, and the expressions of TLR4 mRNA and p38MAPK mRNA in the lung tissues were significantly increased in model group.Compared with those in model group, the pathohistological damages in lung and ileum tissues were ameliorated, the levels of TNF-α and IL-6 in BALF, the MPO activity of lung and intestinal tissues and the expressions of TLR4 mRNA and p38MAPK mRNA in the lung tissues were lower in the rats of drainage group [TNF-α in BALF (ng/L): 858.55±27.16 vs. 1 680.58±105.62; IL-6 in BALF (ng/L): 0 vs. 484.71±5.43; MPO activity of lung (U/g): 0.95±0.13 vs. 1.36±0.11; MPO activity of ileum tissues (U/g): 0.75±0.13 vs. 1.30±0.16; TLR4 mRNA: 0.21±0.11 vs. 0.69±0.13, p38MAPK mRNA: 0.21±0.13 vs. 0.47±0.09; allP < 0.05].Conclusion Mesenteric lymph drainage can alleviate acute lung injury in rats with bowel repletion, and its mechanism may be related to the reduction of the expressions of TLR4 mRNA and p38MAPK mRNA and the release of TNF-α and IL-6 in lung tissues.

Intestinal lymph drainage reduces lung tissue cell apoptosis of hemorrhagic shock rats / 中国基层医药

ObjectiveTo explore the effect of the intestinal lymph drainage on lung tissue cell apoptosis in rats with hemorrhagic shock after resuscitation,rich ALI intestinal lymphatic pathway theory.MethodsTUNEL method was used to determine the apoptosis of lung tissue cells,the brown nuclei were apoptotic cells.SABC was used to determine Bcl-2 and Bax protein expression.ResultsThe shock group and shock + drainage group lung tissue cell apoptosis rate were significantly higher than those in the sham operation group,but the shock + drainage group,the apoptosis of lung tissue cells was significantly lower than the shock group.Sham operation group showed Bcl-2,Bax protein of expression; In shock group,lung tissue cell Bcl-2 expression was significantly lower than the sham operation group,the Bax expression was significantly higher than that in the sham operation group; In shock + drainage group,lung tissue cells shored enhanced expression of Bcl-2,Bax expression was reduced,and the shock + drainage group lung tissue cell Bcl-2 expression was significantly higher than that in the shock group,the expression of Bax was significantly lower than the shock group.ConclusionThe excessive apoptosis of lung tissue cells was one of the mechanisms of lung injury after shock.Intestinal lymph drainage could reduce lung tissue cell apoptosis,the mechanism invdved the regulation of Bcl-2/Bax protein expression.

Effect of ω-3 polyunsaturated fatty acids on intestinal ischemiareperfusion injury and lymphatic duct ligation / 中华临床营养杂志

Objective To investigate the effect of intestinal lymphatic duct ligation and ω-3 polyun saturated fatty acids on intestinal and distant organ in intestinal ischemia-reperfusion injury. Methods Totally 40Sprague-Dawley (SD) male rats (SPF grade)after gastrostomy were equally randomized into sham group (Sham), enteral nutrition (EN) group, enteral nutrition and lymphatic duct ligation (EN + L) group, ω-3 polyunsaturated fatty acids (ω-3PUFA) group, and ω-3PUFA and lymphatic duct ligation (ω-3PUFA + L) group. After 7 days of nutritional intervention, rats were subjected to 60 minutes of intestinal ischemia, ischemia plus mesenteric lymph duct ligation, or sham procedures. After 3 days of continuous nutrition intervention using the original nutrient, lymph nodes, lung, intestine, liver, and blood specimens were harvested. Intestinal permeability and morphology, results of bacterial cultures, and serum cytokines were observed or detected. Result After 3 days of intestinal ischemia-reperfusion (I/R), the body weights of rats in EN group significantly decreased when com pared with the pre-I/R levels (P ＜ 0.05), while the body weights of rats in EN + L group were significantly lower than those in ω-PUFA group and ω-PUFA + L group (P ＜ 0. 05). After one day of intestinal ischemia-reperfusion (I/R), the L/M significantly increased in each group (P ＜0.05 or P ＜0. 01). After 3 days of intestinal ischemiareperfusion (I/R) , the L/M were significantly lower than the level one day after ischemia- reperfusion in EN + L group, ω-PUFA group, and ω-PUFA + L group (P ＜ 0.05). The L/M in EN group and EN + L group were significantly higher than that in ω-PUFA + L group (P ＜ 0. 05). The mucosa thickness and villus height of jejunum in ω-PUFA group and ω-PUFA + L group were significantly higher than those in Sham group, EN group, and EN + L group (P ＜ 0. 01 or P ＜ 0. 05). The mucosa thickness and villus height of ileum in ω-PUFA group and ω-PUFA +L group were also significantly higher than those in EN group (P ＜ 0.05). In ω-PUFA + L group, the serum endotoxin level and tumor necrosis factor-α level were significantly lower than those in EN group (P ＜ 0.05), interleukin (IL) -6 level was significantly lower than that in the ω-PUFA group (P ＜ 0.05), and IL-1 β level was significantly lower than those in other groups (P ＜ 0. 05). In EN group, the lung cell apoptosis index was significantly higher than those in other groups (P ＜ 0.05)and the levels of inducible nitric oxide synthase (iNOS)and myeloperoxidase (MPO) were significantly higher than those in ω-PUFA + L group (P ＜ 0. 05). The level of iNOS was also significantly higher in EN + L group than that in ω-PUFA + L group (P ＜ 0.05). Conclusions Sixty minutes of intestinal ischemia can cause intestinal injury, intestinal barrier dysfunction, and increased permeability of intestine. After 72 h of reperfusion, the intestinal injury can be partially recovered and the permeability can be lower than the post-ischemia level; however, bacterial endotoxin translocation and lung apoptotic cells still exist. Intestinal lymphatic ligation can alleviate the lung damage, promote repair of intestinal mucosa, reduce endotoxin translocation, and attenuate the systemic inflammatory response. EN added with ω-3PUFA is remarkably superior to conventional EN.

Mesenteric lymph duct ligation relives hepatic injury subjected to two-hit in rats / 基础医学与临床

Objective To explore the mechanism of mesenteric lymph duct ligation relieving hepatic injury in rats by two-hit of hemorrhage and LPS.Methods Forty-five Wistar rats were divided into three groups: ligation group,non-ligation group and sham group,and the two-hit model was established by hemorrhage and LPS,mesenteric lymph was blocked by ligating mesenteric lymph duct in ligation group.After 24 hours of operation,took out the liver for pathological section,and the hepatocellular apoptosis rate was determined by method of TUNEL,the expression of BCL-2 and BAX protein was determined by immunohistochemical test.At the same time,taking out liver for homogenate of 10 percent,the activity of MPO and ATPase and the contents of TNF-? and IL-6 were determined in hepatic homogenate.Results After two-hit,the hepatocellular apoptosis rate and expression of BAX protein in non-ligation group were significantly increased as compared with sham group and ligation group,and expression of BCL-2 protein was significantly lower.The contents of MPO,TNF-? and IL-6 in hepatic homogenate of non-ligation group were significantly increased than that of sham group,and the activity of ATPase in hepatic homogenate was significantly lower.But the ATPase in hepatic homogenate of ligation group were significantlyincreased and MPO,TNF-? and IL-6 in hepatic homogenate of ligation group were significantly lower as compared with non-ligation group.Conclusion The mechanism of mesenteric lymph duct ligation relieving hepatic injury of rats was related to the mesenteric lymph blockage reduces the TNF-? and IL-6 and improves the expression of BCL-2 protein and the activity of ATPase in liver.