Protective effect of clodronate-containing liposomes on intestinal mucosal injury in rats with severe acute pancreatitis

BACKGROUND: Severe acute pancreatitis (SAP) can result in intestinal mucosal injury. This study aimed to demonstrate the protective effect of clodronate-containing liposomes on intestinal mucosal injury in rats with SAP. METHODS: Liposomes containing clodronate or phosphate buffered saline (PBS) were prepared by the thin-film method SAP models were prepared by a uniform injection of sodium taurocholate (2 mL/kg body weight) into the subcapsular space of the pancreas. Sprague-Dawley rats were randomly divided into a control group (C group), a SAP plus PBS-containing liposomes group (P group) and a SAP plus clodronate-containing liposomes group (T group). At 2 and 6 hours after the establishment of SAP models, 2 mL blood samples were taken from the superior mesenteric vein to measure the contents of serum TNF-α and IL-12. Pathological changes in the intestine and pancreas were observed using hematoxylin and eosin staining, while apoptosis was detected using TUNEL staining. In addition, the macrophage markers cluster of differentiation 68 (CD68) in the intestinal tissue was assessed with immunohistochemistry. RESULTS: At the two time points, the levels of TNF-α and IL-12 in the P group were higher than those in the C group (P<0.05) Compared with the P group, the levels of TNF-α and IL-12 decreased in the T group (P<0.05). The pathological scores of the intestinal mucosa and pancreas in the T group were lower than those of the P group. In the T group, large numbers of TUNEL-positive cells were observed, but none or few in the C and P groups. The number of CD68-positive macrophages decreased in the T group.CONCLUSIONS: Clodronate-containing liposomes have prote- ctive effects against intestinal mucosal injury in rats with SAP. The blockade of macrophages may provide a novel therapeutic strategy in SAP.

Preventive effect of tetramethylpyrazine on intestinal mucosal injury in rats with acute necrotizing pancreatitis

AIM： To evaluate the role of microcirculatory disorder （MCD） and the therapeutic effectiveness ;of tetramethylpyrazine （TMP） on intestinal mucosa injury in rats with acute necrotizing pancreatitis （ANP）.METHODS： A total of 192 Sprague-Dawley rats were randomly divided into three groups： normal control group （C group）, ANP group not treated with TMP （P group）, ANP group treated with TMP （T group）. An ANP model was induced by injection of 50 g/L sodium taurocholate under the pancreatic membrane （4 mL/kg）. C group received isovolumetric injection of 9 g/L physiological saline solution using the same method. T group received injection of TMP （10 mL/kg） via portal vein. Radioactive biomicrosphere technique was used to measure the blood flow at 0.5, 2, 6 and 12 h after the induction of ANP. Samples of pancreas, distal ileum were collected to observe pathological changes using a validated histology score. Intestinal tissues were also used for examination of myeloperoxidase （MPO） expressed intraceUularly in azurophilic granules of neutrophils.RESULTS： The blood flow was significantly lower in P group than in C group （P 〈 0.01）. The pathological changes were aggravated significantly in P group. The longer the time, the severer the pathological changes. The intestinal MPO activities were significantly higher in P group than in C group （P 〈 0.01）. The blood flow of intestine was significantly higher in T group than in P group after 2 h （P 〈 0.01）. The pathological changes were alleviated significantly in T group. MPO activities were significantly lower in T group than in P group （P 〈 0.01 or P 〈 0.05）. There was a negative correlation between intestinal blood flow and MPO activity （r = -0.981, P 〈 0.01） as well as between intestinal blood flow and pathologic scores （r = -0.922, P 〈 0.05）.CONCLUSION： MCD is an important factor for intestinal injury in ANP. TMP can ameliorate the condition of MCD and the damage to pancreas and intestine.

Over-starvation aggravates intestinal injury and promotes bacterial and endotoxin translocation under high-altitude hypoxic environment

AIM:To study whether over-starvation aggravates intestinal mucosal injury and promotes bacterial and endotoxin translocation in a high-altitude hypoxic environment.METHODS:Sprague-Dawley rats were exposed to hy-pobaric hypoxia at a simulated altitude of 7000 m for 72 h.Lanthanum nitrate was used as a tracer to detect intestinal injury.Epithelial apoptosis was observed with terminal deoxynucleotidyl transferase dUTP nick end labeling staining.Serum levels of diamino oxidase(DAO),malondialdehyde(MDA),glutamine(Gln),superoxide dismutase(SOD) and endotoxin were measured in intestinal mucosa.Bacterial translocation was detected in blood culture and intestinal homogenates.In addition,rats were given Gln intragastrically to observe its protective effect on intestinal injury.RESULTS:Apoptotic epithelial cells,exfoliated villi and inflammatory cells in intestine were increased with edema in the lamina propria accompanying effusion of red blood cells.Lanthanum particles were found in the intercellular space and intracellular compartment.Bacterial translocation to mesenteric lymph nodes(MLN) and spleen was evident.The serum endotoxin,DAO and MDA levels were significantly higher while the serum SOD,DAO and Gln levels were lower in intestine(P< 0.05).The bacterial translocation number was lower in the high altitude hypoxic group than in the high altitude starvation group(0.47±0.83 vs 2.38±1.45,P<0.05).The bacterial translocation was found in each organ,especially in MLN and spleen but not in peripheral blood.The bacterial and endotoxin translocations were both markedly improved in rats after treatment with Gln.CONCLUSION:High-altitude hypoxia and starvation cause severe intestinal mucosal injury and increase bacterial and endotoxin translocation,which can be treated with Gln.

Dynamic changes of IL-2/IL-10, sFas and expression of Fas in intestinal mucosa in rats with acute necrotizing pancreatitis

AIM:To investigate dynamic changes of serum IL-2, IL-10, IL-2/IL-10 and sFas in rats with acute necrotizing pancreatitis. To explore the expression of Fas in intestinal mucosa of rats with acute necrotizing pancreatitis (ANP). METHODS:A total of 64 Sprague-Dawley (SD) rats were randomly divided into two groups:normal control group (C group), ANP group (P group). An ANP model was induced by injection of 50 g/L sodium taurocholate under the pancreatic membrane. Normal control group received isovolumetric injection of 9 g/L physiological saline solution using the same method. The blood samples of the rats in each group were obtained via superior mesenteric vein to measure levels of IL-2, IL-10, sFas and calculate the value of IL-2/IL-10. The levels of IL-2, IL-10 and sFas were determined by ELISA. The severity of intestinal mucosal injury was evaluated by pathologic score. The expression of Fas in intestinal mucosal tissue was determined by immunohistochemistry staining. RESULTS:Levels of serum IL-2 were significantly higher in P group than those of C group (2.79 ± 0.51 vs 3.53 ± 0.62, 2.93 ± 0.89 vs 4.35 ± 1.11, 4.81 ± 1.23 vs 6.94 ± 1.55 and 3.41 ± 0.72 vs 4.80 ± 1.10, respectively, P < 0.01, for all) and its reached peak at 6 h. Levels of serum IL-10 were significantly higher in P group than those of C group at 6 h and 12 h (54.61 ± 15.81 vs 47.34 ± 14.62, 141.15 ± 40.21 vs 156.12 ± 43.10, 89.18 ± 32.52 vs 494.98 ± 11.23 and 77.15 ± 22.60 vs 93.28 ± 25.81, respectively, P < 0.01, for all). The values of IL-2/IL-10 were higher significantly in P group than those of C group at 0.5 h and 2 h (0.05 ± 0.01 vs 0.07 ± 0.02 and 0.02 ± 0.01 vs 0.03 ± 0.01, respectively, P < 0.01, for all), and it were significantly lower than those of C group at 6 h (0.05 ± 0.02 vs 0.01 ± 0.01, P < 0.01) and returned to the control level at 12 h (0.04 ± 0.01 vs 0.05 ± 0.02, P > 0.05). In sFas assay, there was no significant difference between P group and C group (3.16 ± 0.75 vs 3.31 ± 0.80, 4.05 ± 1.08 vs 4.32 ± 1.11, 5.93 ± 1.52 vs 5.41 ± 1.47 and 4.62 ± 1.23 vs 4.44 ± 1.16, respectively, P > 0.05, for all). Comparison of P group and C group, the pathological changes were aggravated significantly in P group. Immunohistochemistry staining show the expression of Fas was absent in normal intestinal tissues, however, it gradually increased after induction of pancreatitis in intestinal tissue, then reached their peaks at 12 h.CONCLUSION:Fas were involved in the pathogenesis of pancreatitis associated intestinal injury. The mechanisms of Fas may be associated to Fas mediated T helper cell apoptosis.

Pretreatment of cromolyn sodium prior to reperfusion attenuates early reperfusion injury after the small intestine ischemia in rats

AIM: To investigate the effects of Cromolyn Sodium (CS) pretreated prior to reperfusion on the activity of intestinal mucosal mast cells (IMMC) and mucous membrane of the small intestine in ischemia-reperfusion (IR) injury of rats. METHODS: Thirty-two Sprague-Dawley (SD) rats were randomly divided into four groups: sham group (group S), model group (group M), high and low dosage of CS groups, (treated with CS 50 mg/kg or 25 mg/kg, group C1 and C2). Intestinal IR damage was induced by clamping the superior mesenteric artery for 45 min followed by reperfusion for 60 min. CS was intravenouly administrated 15 min before reperfusion. Ultrastructure and counts of IMMC, intestinal structure, the expression of tryptase, levels of malondisldehyde (MDA), TNF-α, histamine and superoxide dismutase (SOD) activity of the small intestine were detected at the end of experiment. RESULTS: The degranulation of IMMC was seen in group M and was attenuated by CS treatment. Chiu’s score of group M was higher than the other groups. CS could attenuate the up-regulation of the Chiu’s score, the levels of MDA, TNF-α, and expression of tryptase and the down-regulation of SOD activity and histamine concentration. The Chiu’s score and MDA content were negatively correlated, while SOD activity was positively correlated to the histamine concentration respectively in the IR groups. CONCLUSION: Pretreated of CS prior to reperfusion protects the small intestine mucous from ischemia- reperfusion damage, the mechanism is inhibited IMMC from degranulation.

TNF-α and plasma D(-)-lactate levels in rats after intestinal ischemia and reperfusion

Objective To study the potential role of tumor necrosis factor-α (TNF-α) induction in the development of mucosal barrier dysfunction in rats caused by acute intestinal ischemia-reperfusion injury, and to examine whether pretreatment with monoclonal antibody against TNF-α (TNF-α MoAb) would affect the release of D(-)-lactate after local gut ischemia followed by reperfusion. Methods Anesthetized Sprague-Dawley rats underwent superior mesenteric artery occlusion for 75 min followed by reperfusion for 6 hr. The rats were treated intravenously with either TNF-α MoAb (20 mg/kg) or albumin (20 mg/kg) 30 min prior to the onset of ischemia. Plasma D(-)-lactate levels were measured in both the portal and systemic blood by an enzymatic spectrophotometric assay. Intestinal TNF-αmRNA expression as well as protein levels were also measured at various intervals. In addition, a postmortem examination was performed together with a macropathological evaluation based on a four-grade scoring system.Results Intestinal ischemia resulted in a significant elevation in D(-)-lactate levels in the portal vein blood in both the control and treatment groups ( P ＜0.05). However, animals pretreated with TNF-α MoAb at 6 hr after reperfusion showed significant attenuation of an increase in both portal and systemic D(-)-lactate levels when compared with those only receiving albumin (P ＜ 0.05). In the control animals, a remarkable rise in intestinal TNF-α level was measured at 0.5 hr after clamp release ( P ＜ 0.01); however, prophylactic treatment with TNF-α MoAb completely annulled the increase of local TNF-α levels seen in the control animals. Similarly, after anti-TNF-α MoAb administration, intestinal TNF-α mRNA expression was markedly inhibited, which showed significant differences when compared with the control group at 0.5 hr, 2 hr and 6 hr after the release of occlusion ( P ＜ 0.05-0.01 ). In addition, the pathological examination showed marked intestinal lesions that formed during ischemia, which were much worse upon reperfusion,particularly at the 6 hr time point. These acute injuries were obviously attenuated in animals receiving TNF-α MoAb.Conclusions It appeared that acute intestinal ischemia was associated with failure of the mucosal barrier, resulting in increased plasma D(-)-lactate levels in both portal and systemic blood. These results suggest that TNF-α appears to be involved in the development of local damage associated with intestinal ischemic injury. Moreover, prophylactic treatment with TNF-α MoAb exerts preventive effects on ischemia/ reperfusion-induced circulating D (-)-lactate elevation and gut injury. ( J Geriatr Cardiol 2004;1(2):119-124. )

Effect of Weikang Capsule(胃康胶囊)on Aspirin-Related Gastric and Small Intestinal Mucosal Injury

Objective To investigate the effects of Weikang Capsule(胃康胶囊,WKC)on aspirin-related gastric and small intestinal mucosal injury by magnetically controlled capsule endoscopy(MCCE).Methods Patients taking enteric-coated aspirin aged 40-75 years were enrolled in Beijing Anzhen Hospital,Capital Medical University from January 2019 to December 2019.The patients continued taking aspirin Tablet(100 mg per day)and underwent MCCE before and after 1-month combined treatment with WKC(0.9 g per time orally,3 times per day).The gastrointestinal symptom score,gastric Lanza score,the duodenal,jejunal and ileal mucosal injury scores were used to evaluate the gastrointestinal injury before and after treatment.Adverse events including nausea,vomiting,abdominal pain,abdominal distension,abdominal discomfort,dizziness,or headache during MCCE and combined treatment were observed and recorded.Results Twenty-two patients(male/female,13/9)taking enteric-coated aspirin aged 59.5±11.3 years with a duration of aspirin use of 28.0(1.0,48.0)months were recruited.Compared with pre-treatment,the gastrointestinal symptom rating scale scores,gastric Lanza scores,and duodenal mucosal injury scores were significantly reduced after 1-month WKC treatment(P<0.05),and jejunal and ileal mucosal injury scores showed no obvious change.No adverse events occurred during the trial.Conclusions WKC can alleviate gastrointestinal symptoms,as well as gastric and duodenal mucosal injuries,in patients taking enteric-coated aspirin;it does not aggravate jejunal or ileal mucosal injury,which may be an effective alternative for these patients(Clinical trial registry No.ChiCTR1900025451).

Evaluating a novel protective agent against radiation-induced acute intestinal injury

Objective:To develop and synthesize a novel derivative of ethyl pyruvate,named TZC02,and investigate its radioprotective effects against ionizing radiation(IR)-induced intestinal injury in mice.Methods:Male C57BL/6J mice weighing(20±2)g in the survival experiment were randomly divided into five groups(n=10 in each):control group,IR group,IR+TZC02(50 mg/kg)group,IR+TZC02(100 mg/kg)group,and IR+TZC02(200 mg/kg)group.Mice's survival rates were monitored for 7 d.In other experiments,the male mice were randomly divided into three groups(n=5 per group):control group,IR group,and IR+TZC02(100 mg/kg)group.TZC02 was intragastrically administered 1 h before 12 Gy abdominalγ-ray irradiation(ABI)and 24 h,48 h after irradiation,respectively.Three days after IR exposure,small intestinal tissues were collected and the number of small intestinal crypts was determined using hematoxylin&eosin(H&E)staining.Immunohistochemical analysis was used to assess the regenerative capacity of the small intestine(SI)and radiation-induced damage,stemness markers or DNA repair surrogates,including Ki67,lysozyme,and villus.The expressions of histone H2AX phosphorylation(γH2AX)and caspase-3 were evaluated through immunofluorescence analyses.Additionally,in vitro cultured small intestinal organoids were employed to investigate the effects of TZC02 on SI regeneration after irradiation.Results:The administration of TZC02 significantly improved the 7 d-survival rate of mice exposed to 12 Gy ABI(P<0.05).Compared to the IR group,TZC02 treatment attenuated the decrease of SI Ki67-positive cells[(59.60±6.33)vs.(37.70±7.82),t=11.89,P<0.0001]and Paneth cells[(9.90±1.37)vs.(5.50±1.71),t=6.02,P<0.001]in five crypts,and reduced structural damage to the SI[villus height,(349.49±60.17)μm vs.(294.72±40.09)μm;t=3.39;P<0.05].TZC02 also significantly decreased the crypt apoptosis detected by caspase-3[(10.75±1.26)vs.(29.83±2.56),t=13.39,P<0.0001]and DNA damage detected by gH2AX[(10.40±1.14)vs.(29.60±2.70),t=10.13,P<0.0001].The organoid survival 7 d post-irradiation further confirmed the protective effects of TZC02(area of organoids,(0.119±0.081)mm^(2)vs.(0.080±0.037)mm^(2);t=2.30;P<0.05).Conclusions:This study demonstrate that TZC02 can offer effective protection against IR-induced intestinal injury,suggesting its potential as a promising protective compound for patients treated with radiotherapy.

谷氨酰胺治疗服用阿司匹林患者小肠黏膜损伤的随机对照研究

目的探讨谷氨酰胺对正在服用阿司匹林患者相关小肠黏膜损伤的治疗作用。方法选取2022年1月至2024年1月于首都医科大学附属北京安贞医院就诊正在服用阿司匹林,经胶囊内镜筛查有小肠黏膜损伤的患者为研究对象,采用计算机随机序列法分为干预组和对照组,干预组继续服用阿司匹林的同时加用复方谷氨酰胺肠溶胶囊治疗,对照组仅继续服用阿司匹林,不采取其他针对阿司匹林损伤的干预治疗,治疗2个月后再次行胶囊内镜检查。采用小肠内镜5级评分标准评价2组患者小肠黏膜损伤情况,研究的主要终点是2组患者小肠黏膜损伤评分的变化。结果成功入组并完成全部临床试验的共有43例患者,其中干预组22例,对照组21例。干预组治疗后小肠黏膜损伤评分较治疗前明显减低[1.0(0.0,1.3)分比2.0(2.0,3.0)分,P<0.001],对照组治疗前后小肠黏膜损伤评分比较差异无统计学意义[2.0(2.0,2.0)分比2.0(2.0,2.0)分,P=0.317]。干预组较对照组小肠黏膜损伤评分减低更多[1.0(1.0,2.0)分比0.0(0.0,0.0)分,P<0.001]。结论谷氨酰胺可以用于正在服用阿司匹林的患者以减轻相关小肠黏膜损伤。

肠脂肪酸结合蛋白在肠黏膜屏障损伤相关疾病中的作用研究进展

肠道是人体持续暴露于外部环境的最大器官之一,发挥着重要的物理及化学屏障作用保护人体免受致病菌侵害,肠黏膜屏障作为肠道的重要组成部分,其完整性对维持肠道微生态平衡起关键作用,肠黏膜屏障破坏可导致肠道菌群失调及易位,与多种疾病的发生发展相关。肠脂肪酸结合蛋白(intestinal fatty acid binding protein,I‑FABP)是脂肪酸结合蛋白家族中的一种,对肠道损伤具有特异性,在肠道发生损伤、缺血、肠黏膜屏障损伤及通透性增加时会迅速释放入血液循环,I‑FABP水平升高是反映肠道损伤的重要标志物,临床上很多疾病常合并肠道功能障碍,本文主要对I‑FABP的结构特征、功能及其与肠黏膜屏障相关疾病的关系进行综述。

苦参碱调节IL-6/STAT3信号通路对炎症性肠病大鼠肠黏膜损伤的影响

【目的】观察苦参碱调节白细胞介素6(IL-6)/信号转导子和转录激活子3(STAT3)信号通路对炎症性肠病(IBD)大鼠肠黏膜损伤的影响。【方法】以三硝基苯磺酸(TNBS)结肠灌注法构建IBD大鼠模型,随机分为模型组,苦参碱低、高剂量组,苦参碱高剂量+colivelin(STAT3激活剂)组,每组10只;再选10只大鼠结肠灌注等体积生理盐水作为正常组。经苦参碱和colivelin处理后,检测各组大鼠体质量和疾病活动指数(DAI),采用苏木素-伊红(HE)染色法检测大鼠结肠黏膜组织病理学变化,透射电镜观察大鼠结肠黏膜组织超微结构变化,采用酶联免疫吸附分析(ELISA)和比色法分别检测大鼠血清和结肠黏膜组织白细胞介素6(IL-6)、C-反应蛋白(CRP),超氧化物歧化酶(SOD)、丙二醛(MDA)水平,采用蛋白免疫印迹(Western Blot)法检测大鼠结肠黏膜组织IL-6/STAT3通路相关蛋白表达。【结果】与正常组比较,模型组大鼠结肠黏膜组织发生严重病理损伤且其超微结构受损明显,DAI,结肠黏膜组织病理评分,血清和结肠黏膜组织CRP、IL-6、MDA水平,结肠黏膜组织IL-6蛋白表达及p-STAT3/STAT3比值显著升高(P<0.05),体质量、血清和结肠黏膜组织SOD水平显著降低(P<0.05);与模型组比较,苦参碱低、高剂量组大鼠结肠黏膜组织病理损伤及其超微结构受损均减轻,DAI,结肠黏膜组织病理评分,血清和结肠黏膜组织CRP、IL-6、MDA水平,结肠黏膜组织IL-6蛋白表达及p-STAT3/STAT3比值均降低(P<0.05),体质量、血清和结肠黏膜组织SOD水平均升高(P<0.05),且呈剂量依赖性;与苦参碱高剂量组比较,苦参碱高剂量+colivelin组大鼠结肠黏膜组织病理损伤及其超微结构受损加重,DAI,结肠黏膜组织病理评分,血清和结肠黏膜组织CRP、IL-6、MDA水平,结肠黏膜组织IL-6蛋白表达及p-STAT3/STAT3比值升高(P<0.05),体质量、血清和结肠黏膜组织SOD水平降低(P<0.05)。【结论】苦参碱可通过阻止IL-6/STAT3信号通路激活降低炎症及氧化应激水平,进而减轻IBD大鼠肠黏膜损伤。

解毒护肝通络方对药物性肝损伤小鼠的保护作用及其对肠道菌群和肠黏膜屏障的影响

目的探讨解毒护肝通络方对药物性肝损伤(DILI)小鼠的保肝作用及其机制。方法将C57BL/6N小鼠60只,随机分为正常组、对乙酰氨基酚(APAP)组、阳性药(水飞蓟宾)组及解毒护肝通络方高、中、低剂量组。上午采用APAP药液灌胃复制DILI小鼠模型,下午灌胃相应药物治疗,持续14天。小鼠处死后,收集血清、肝脏、回肠、结肠组织以及粪便用于后续指标检测。结果与正常组比,APAP组小鼠血清中谷丙转氨酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate aminotransferase,AST)、直接胆红素(direct bilirubin,DBIL)、总胆红素(total bilirubin,TBIL)和脂多糖(lipopolysaccharide,LPS)的含量或活性均显著升高(P<0.01),肝脏、回肠和结肠组织均出现病理损伤,小肠绒毛高度、隐窝深度、菌群丰富度和多样性、肠道组织闭合蛋白(Occludin)和闭锁小带蛋白-1(ZO-1)表达水平以及拟杆菌门和S24-7相对丰度均显著降低(P<0.01,P<0.05),厚壁菌门、厚壁菌门与拟杆菌门比值(F/B)和毛螺菌科相对丰度升高(P<0.01,P<0.05)。与APAP组比,解毒护肝通络方能显著改善小鼠肝功能以及肝脏、回肠和结肠组织病理损伤,降低血清LPS含量(P<0.01),升高小肠绒毛高度、隐窝深度、菌群丰富度和多样性、肠道组织中occludin和ZO-1蛋白表达水平以及拟杆菌门、S24-7的相对丰度(P<0.01,P<0.05),降低F/B、厚壁菌门和毛螺菌科相对丰度(P<0.01)。结论解毒护肝通络方可能通过恢复肠道菌群失调,保护肠黏膜屏障,降低肠道通透性,进而达到保肝的目的。

低压低氧致小鼠胃肠应激损伤的研究

目的通过低压氧舱模拟高原缺氧环境,探究缺氧不同时间对小鼠胃肠应激损伤的影响。方法根据低压低氧不同时间设置正常对照组、低压低氧1、3、5天模型组,低压低氧处理完成后检测小肠推进率、内脏敏感性变化及胃肠道病理损伤程度;酶联免疫吸附法(ELISA)检测胃肠道IL-1β、IL-6、TNF-α含量变化,肠组织胃肠激素及肠黏膜损伤标志物二胺氧化酶、D-乳酸含量变化;Western blot实验检测肠组织紧密连接蛋白及紧密连接损伤通路相关蛋白表达变化。结果与对照组相比,模型组小鼠小肠推进率加快,肠道敏感性升高;肠黏膜损伤标志物表达升高;肠组织抑制胃肠蠕动激素降低,促胃肠蠕动激素升高;紧密连接蛋白ZO-1、claudin-1、occludin及紧密连接损伤相关通路蛋白VASP表达降低,紧密连接损伤相关通路中的NF-κB、HIF-1α、VEGF表达升高;胃肠组织IL-1β、IL-6、TNF-α表达升高;胃组织胃蛋白酶含量升高;胃肠组织1天组损伤较轻,3天、5天组损伤较重;结论在低氧低氧环境中不同时间小鼠胃肠发生应激损伤,暴露3天时胃肠功能损伤较为严重,其中肠损伤机制可能与NF-κB/HIF-1α/VEGF/VASP信号通路被激活有关。

大承气汤鼻饲对重症胰腺炎合并肺损伤患者氧合指数、炎症因子及肠黏膜屏障功能的影响

目的:探讨大承气汤联合西药对重症急性胰腺炎(SAP)合并肺损伤患者进行治疗的效果,及对氧合指数、炎症因子、肠黏膜屏障功能的影响。方法:选用2022年10月~2023年7月本院收治的90例SAP合并肺损伤患者为研究对象,随机分为两组(中药组45例、常规组45例)。常规组采用奥曲肽进行治疗,中药组在常规组的基础上联合大承气汤进行治疗。比较两组治疗后总有效率、动脉血气指标、临床症状缓解时间、住院时间、各项炎症因子水平、T细胞亚群水平、肠黏膜屏障功能指标等。结果:治疗后,常规组总有效率75.56%较中药组总有效率95.56%明显更低(P<0.05);两组动脉血气指标均显著升高(P<0.05),且中药组血氧分压(PaO_(2))、氧合指数(OI)水平明显更高(P<0.05);两组各症状消失时间、住院时间显著缩短(P<0.05),且中药组较常规组更短(P<0.05);两组肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、CD3+、CD4+、CD8+水平均显著降低(P<0.05),且中药组下降幅度更大(P<0.05);两组内毒素(ET)、二胺氧化酶(DAO)、D-乳酸水平较治疗前明显下降(P<0.05),且中药组各项肠黏膜屏障功能指标改善程度更高(P<0.05)。结论:对SAP合并肺损伤患者采用西药加大承气汤鼻饲治疗,效果较为理想,临床应用价值较高。

补中益气汤对高原低氧大鼠肠道屏障的保护作用研究

目的评价补中益气汤对高原低氧暴露大鼠小肠黏膜损伤的调控作用。方法选用7周龄雄性Wistar大鼠18只,适应性饲养7天后采用随机数字表法分成对照组、低氧暴露组、补中益气汤干预组,每组6只。各组动物预先灌胃7天后,除对照组大鼠外,其余采用低压氧舱模拟海拔高度6000米进行高原低氧暴露诱导肠黏膜损伤。记录大鼠体质量和摄食量的变化;苏木精—伊红染色观察空肠组织形态变化,生化法检测大鼠血清髓过氧化物酶(myeloperoxidase,MPO)和二胺氧化酶(diamine oxidase,DAO)活性,酶联免疫吸附法检测胃泌素(gastrin,GAS)水平;二氢乙啶(dihydroethidium,DHE)染色检测空肠组织活性氧(reactive oxygen species,ROS)水平;硫代巴比妥酸法和比色法检测血清超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-PX)和丙二醛(malondialdehyde,MDA)水平;通过免疫蛋白印迹实验检测空肠组织中紧密连接蛋白闭锁蛋白、闭锁小带-1蛋白及闭合蛋白的表达情况。结果与对照组相比,低氧暴露组大鼠体质量和摄食量明显减少(P<0.01),空肠绒毛高度和隐窝深度的比值显著下降(P<0.01);大鼠血清中肠道屏障损伤指标DAO和MPO水平显著增高及GAS分泌降低(P<0.01);大鼠血清中脂质过氧化水平(MDA)升高,抗氧化能力(SOD、GSH-PX)显著降低(P<0.01);空肠组织紧密连接蛋白闭锁蛋白、闭锁小带-1蛋白及闭合蛋白表达水平降低(P<0.05)。与低氧暴露组相比,补中益气汤组绒毛长度和隐窝深度有所恢复(P<0.01);大鼠血清中的DAO和MPO含量显著降低,GAS分泌增加(P<0.01);大鼠血清中的SOD、GSH-PX含量明显升高,MDA水平降低(P<0.01);大鼠空肠组织紧密连接蛋白表达水平显著升高,差异均有统计学意义(P<0.01)。结论补中益气汤可显著改善高原低氧暴露导致的肠黏膜损伤,其可能与改善机体氧化应激损伤,提高肠黏膜紧密连接蛋白的表达有关。

Akkermansia muciniphila上清通过抑制内质网应激改善小肠黏膜损伤

目的:探究Akkermansia muciniphila(AKK)上清对非甾体类抗炎药(NSAIDs)导致小肠黏膜损伤的干预效果及作用机制。方法:将18只C57BL/6小鼠(6~8周龄)采用随机数字表法随机分为3组:正常对照组(CON组)、造模组(INDO组)、干预组(INDO+AKK组),每组6只。INDO组使用吲哚美辛构建NSAIDs小肠损伤模型,INDO+AKK组使用AKK上清灌胃。采集各组肠道标本,常规苏木精-伊红(HE)染色观察黏膜屏障形态,免疫组织化学染色法(IHC)观察黏膜屏障指标咬合蛋白(Occuldin)、闭锁小带蛋白(ZO1)、黏蛋白2(MUC2)的表达水平。采用实时荧光定量PCR(qRT-PCR)从mRNA水平验证MUC2、葡萄糖调节蛋白78(GRP78)、X盒结合蛋白1(XBP1s)、C/EBP坏死因子相关蛋白(CHOP)、激活转录因子4(ATF4)、激活转录因子6(ATF6)、真核翻译起始因子2α激酶3(PERK)等基因的表达变化。结果:小肠组织HE染色观察发现,与CON组相比,INDO组出现小肠黏膜结构破坏、绒毛明显减少等特征。INDO+AKK组小肠黏膜损伤减轻、绒毛丧失现象减少、黏膜及其下层的炎症细胞浸润区域也显著减小。IHC分析显示,与INDO组相比,INDO+AKK组Occuldin(F=11.48,P<0.05)、ZO1(F=68.10,P<0.001)、MUC2(F=19.93,P<0.01)表达水平增加。PAS结果显示,INDO+AKK组杯状细胞数量相对于INDO组明显增加(F=205.9,P<0.001)。qRTPCR结果发现,与INDO组相比,INDO+AKK组MUC2的基因(F=23.67,P<0.01)表达水平升高,GRP78(F=7.869,P<0.01)、CHOP(F=11.45,P<0.05)、XBP1s(F=8.344,P<0.05)、ATF4(F=16.37,P<0.001)、ATF6(F=12.99,P<0.001)、PERK(F=11.58,P<0.01)的基因表达水平均下降。结论:AKK上清通过抑制内质网应激,改善NSAIDs诱导的小肠黏膜屏障损伤。

柚皮苷通过P2X7受体抑制非经典焦亡减轻腹腔感染小鼠肠黏膜损伤

目的 探究柚皮苷(naringin, Ng)在腹腔感染(intra-abdominal infection, IAI)小鼠肠黏膜损伤中的治疗作用及其机制。方法 腹腔注射脂多糖(lipopolysaccharide, LPS)建立IAI小鼠肠黏膜损伤模型。64只健康雄性小鼠采用随机数字表法分为4组(n=16):对照组(Control组,0.2 mL PBS),脂多糖组(LPS组,LPS 10 mg/kg),柚皮苷组(Ng组,Ng 50 mg/kg+LPS 10 mg/kg)和BzATP组(BzATP 5 mg/kg+Ng 50 mg/kg+LPS 10 mg/kg)。Ng和BzATP在造模前1 h进行腹腔注射,Control组予以等量PBS。各组于造模24 h随机选6只小鼠获取回肠。HE染色观察回肠黏膜病理学变化和Chiu’s评分;免疫荧光观察肠黏膜P2X7、焦亡效应蛋白GSDMD、巨噬细胞标记物CD68表达和分布情况;Western blot检测回肠组织P2X7和焦亡效应蛋白NLRP3、Caspase11、GSDMD及GSDMD-N表达量;ELISA检测回肠炎症因子IL-1β、IL-18和IL-10水平。记录7 d内小鼠脓毒症评分(mouse sepsis score, MSS)。结果 LPS组与Control组相比,MSS和Chiu’s评分增高(P<0.05),回肠黏膜炎性损伤;肠黏膜P2X7、GSDMD和CD68免疫荧光分布一致且信号增强(P<0.05);促炎因子IL-1β、IL-18水平上升,抗炎因子IL-10水平下降(P<0.05);回肠组织P2X7、NLRP3、Caspase11、GSDMD和GSDMD-N蛋白表达增加(P<0.05)。Ng组与LPS组相比,MSS下降;HE染色和Chiu’s评分表明肠黏膜炎性损伤减轻;P2X7、GSDMD、CD68荧光信号减弱(P<0.05);IL-1β、IL-18水平下降(P<0.05),IL-10水平上升(P<0.05);P2X7、NLRP3、Caspase11、GSDMD和GSDMD-N蛋白表达下降(P<0.05)。BzATP组实验结果与Ng组相反。结论 IAI小鼠肠黏膜P2X7受体表达升高,Caspase11介导的非经典焦亡发生,加重其肠黏膜损伤;Ng可能通过抑制P2X7受体调控非经典焦亡,改善IAI小鼠肠黏膜损伤。

小陷胸汤对5-氟尿嘧啶介导的小鼠肠黏膜损伤的保护作用研究

目的:探究小陷胸汤对5-氟尿嘧啶(5-fluorouracil,5-FU)介导的小鼠肠黏膜损伤的保护作用。方法:将100只SPF级雄性ICR小鼠随机分为正常对照组、模型组、小陷胸汤低、中、高剂量(8.3、16.6、33.2g/kg)组。除正常对照组外,其余各组采用5-FU(30 mg/kg,ip,7 d)诱导肠黏膜损伤小鼠模型。小陷胸汤给药组造模同时按照设定剂量以10 mL·kg^(-1)·d^(-1)灌胃,正常对照组和模型组给予等量蒸馏水,连续处理7 d。采用苏木素-伊红染色法(HE)观察小肠的病理形态学变化并测定肠绒毛高度/隐窝深度(VH:CD)比值;采用阿利新蓝染色法(AB)测定小肠黏膜杯状细胞数量;采用分光光度法检测血清内毒素(ET)、D-乳酸(D-LA)及小肠组织二胺氧化酶(DAO)水平;采用酶联免疫吸附法(ELISA)检测血清肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)水平;采用实时荧光定量聚合酶链式反应法(RT-qPCR)检测小肠组织IL-6、信号转导和转录激活因子3(STAT3)和表皮生长因子受体(EGFR)mRNA表达;采用Western blot法检测小肠组织IL-6、STAT3及EGFR蛋白表达。结果:与正常对照组相比,模型组小鼠体质量显著降低,腹泻评分、粪便质量、粪便含水量及小肠推进率明显升高(P<0.01),小肠黏膜VH:CD比值和杯状细胞数量降低(P<0.01),血清ET和D-LA水平升高,小肠组织DAO水平降低(P<0.05),脾指数和胸腺指数降低(P<0.01),血清IL-6、TNF-α水平升高(P<0.05),小肠组织IL-6、STAT3 mRNA表达升高,EGFR mRNA表达降低(P<0.01);小肠组织IL-6、STAT3蛋白表达升高,EGFR蛋白表达降低(P<0.01)。与模型组相比,小陷胸汤能够明显改善上述指标(P<0.05)。结论:小陷胸汤对5-FU介导的肠黏膜损伤具有保护作用,其作用机制可能与小陷胸汤激活EGFR信号转导,下调IL-6/STAT3信号通路,减轻5-FU诱导的肠黏膜炎症反应,改善免疫功能有关。

黄龙汤联合早期肠内营养对急性胃肠损伤患者肠黏膜屏障功能及营养指标的影响

目的:观察黄龙汤联合早期肠内营养对急性胃肠损伤患者肠黏膜屏障功能及营养指标的影响。方法:将84例急性胃肠损伤患者按照随机数字表法分为对照组和观察组,每组各42例。对照组在应激期内静脉注射奥美拉唑注射液,之后行早期肠内营养方案,观察组在对照组治疗的基础上加用黄龙汤。比较两组患者治疗前后血清瓜氨酸、内毒素、肠型脂肪酸结合蛋白(intestinal fatty acid binding protein,I-FABP)含量、T淋巴细胞水平(CD4^(+)、CD8^(+)及CD4^(+)/CD8^(+))、体质量指数(body mass index,BMI)、血清白蛋白(albumin,ALB)、总蛋白(total protein,PA)、血红蛋白(haemoglobin,Hb)含量,比较两组患者胃潴留、返流误吸、腹胀、腹泻、恶心呕吐等胃肠道耐受情况及预后情况。结果:两组患者治疗后血清瓜氨酸含量高于本组治疗前,血清内毒素和I-FABP含量均低于本组治疗前,治疗后组间比较,差异有统计学意义(P<0.05)。两组患者治疗后CD4^(+)及CD4^(+)/CD8^(+)均高于本组治疗前,CD8^(+)低于本组治疗前,治疗后组间比较,差异有统计学意义(P<0.05)。两组患者治疗后血清ALB、PA、Hb含量均高于本组治疗前,且观察组高于对照组,差异有统计学意义(P<0.05)。观察组胃肠道不耐受症状发生率明显低于对照组,差异有统计学意义(P<0.05)。观察组恢复良好率高于对照组,差异有统计学意义(P<0.05)。结论:黄龙汤联合早期肠内营养可有效改善急性胃肠损伤患者肠黏膜屏障功能,提高机体营养状态和免疫功能,减少胃肠道不耐受情况,改善患者预后。

磁控机器人胶囊内镜在药物相关性小肠黏膜损伤诊断中的应用价值

目的探索磁控机器人胶囊内镜(magnetic control robot capsule endoscopy,MCRCE)在药物相关性小肠黏膜损伤中的诊断价值。方法收集2021年9月至2021年12月共119例于我院接受MCRCE检查的患者,询问患者病史及服用治疗心血管疾患药物史,根据患者服用药物状况分为未服药组和服药组,以调查问卷形式,分别统计两组患者的相关信息,采用GraphPad Prism 8.0.1软件进行t检验和χ2检验。结果两组患者年龄、肠道黏膜病变检出率及病变数量比较,差异均有显著统计学意义(P<0.0001),但性别、BMI、幽门螺杆菌感染及饮食习惯方面差异均无统计学意义(P>0.05)。结论服用抗凝药及降压药会造成肠道黏膜的损伤且更易发生多处损伤,MCRCE对肠黏膜损伤具有良好的诊断价值。