Improvement effects of limonin on intestinal injury and intestinal flora disturbance in rats with ulcerative colitis and its mechanism / 中国药房

OBJECTIVE To investigate the improvement effects of limonin on intestinal injury and intestinal flora disturbance in rats with ulcerative colitis （UC） and its mechanism. METHODS UC rat models were established， and 70 rats with successful modeling were randomly divided into model group， limonin low-， medium-， and high-dose groups （12.5， 25， 50 mg/kg）， and sulfasalazine group （positive control group，500 mg/kg）， with 14 rats in each group. Another 14 rats were selected as the control group. After modeling， each group was given the corresponding drug or equal amount of normal saline， once a day， for 2 weeks. Twenty-four hours after the last administration， the general condition of rats was observed and the body weight was measured， and colon tissue was collected for colonic mucosal damage index （CMDI） scoring； the levels of interleukin-1β （IL-1β）， IL-6 and tumor necrosis factor-α （TNF-α） in colon tissue were detected； the pathological changes of colon tissue were observed； the protein expressions of Claudin-1， Occludin， ZO-1， high mobility group protein B1 （HMGB1） and receptor for advanced glycation end products （RAGE） in colon tissue were detected； fecal 16S rRNA sequencing was used to detect the relative abundance of zhangxiaxia5287@163.com intestinal microbiota in rats. RESULTS Compared with the control group， the rats in the model group were in poor mental state， with darker fur， irritable mood， disordered arrangement of colon glands， inflammatory cell infiltration， cell necrosis and edema； CMDI score， the levels of IL-1β， IL-6 and TNF-α， protein expressions of HMGB1 and RAGE in colon tissue， the relative abundance of Proteobacteria and Bacteroidetes were significantly increased （P＜0.05）； body weight， the protein expressions of Claudin-1， Occludin and ZO-1 in colon tissue， the relative abundance of Firmicutes in the intestine were significantly decreased （P＜0.05）. Compared with the model group， general situation and pathological damage of colonic tissue in limonin groups were improved， the levels of the above indicators were significantly reversed （P＜0.05）， and in a dose-dependent manner （P＜0.05）； there was no significant difference in various indexes between sulfasalazine group and limonin high-dose group （P＞0.05）. CONCLUSIONS Limonin can improve intestinal injury and intestinal flora disturbance in UC model rats， the mechanism of which may be associated with the down-regulation of HMGB1/RAGE signaling pathway.

Research progress in prevention and treatment of radiation-induced intestinal injury by traditional Chinese medicine active components / 中国中药杂志

Radiation-induced intestinal injury(RIII), a common complication of radiotherapy for pelvic malignancies, affects the quality of life and the radiotherapy efficacy for cancer. Currently, the main clinical approaches for the prevention and treatment of RIII include drug therapy, hyperbaric oxygen therapy, and surgical treatment. Among these methods, drug therapy is cost-effective. Traditional Chinese medicine(TCM) containing a variety of active components demonstrates mild side effects and good efficacy in preventing and treating RIII. Studies have proven that TCM active components, such as flavonoids, terpenoids, phenylpropanoids, and alkaloids, can protect the intestine against RIII by inhibiting oxidative stress, regulating the expression of inflammatory cytokines, modulating the mitochondrial apoptosis pathway, adjusting intestinal flora, and suppressing cell apoptosis. These mechanisms can help alleviate the symptoms of RIII. The paper aims to provide a theoretical reference for the discovery of new drugs for the prevention and treatment of RIII by reviewing the literature on TCM active components in the last 10 years.

Jujubae Fructus alleviates intestinal injury caused by toxic medicinals in Shizao Decoction based on correlation between intestinal flora and host metabolism / 中国中药杂志

Genkwa Fols, Kansui Radix, and Euphorbiae Pekinensis Radix in Shizao Decoction(SZD) are toxic to intestinal tract. Jujubae Fructus in this prescription can alleviate the toxicity, but the mechanism is still unclear. Therefore, this study aims to explore the mechanism. To be specific, 40 normal Sprague-Dawley(SD) rats were classified into the normal group, high-dose and low-dose SZD groups, and high-dose and low-dose SZD without Jujubae Fructus(SZD-JF) groups. The SZD groups were given(ig) SZD, while SZD-JF groups received the decoction without Jujubae Fructus. The variation of body weight and spleen index were recorded. The patho-logical changes of intestinal tissue were observed based on hematoxylin and eosin(HE) staining. The content of malondialdehyde(MDA) and glutathione(GSH) and activity of superoxide dismutase(SOD) in intestinal tissue were measured to evaluate the intestinal injury. Fresh feces of rats were collected to detect intestinal flora structure by 16S ribosomal RNA gene(16S rDNA) sequencing technology. The content of fecal short chain fatty acids and fecal metabolites was determined by gas chromatography-mass spectrometer(GC-MS) and liquid chromatography-mass spectrometer ultra-fast liquid chromatography-quadrupole-time-of-flight mass spectrometer(UFLC-Q-TOF-MS), separately. Spearman's correlation analysis was employed to analyze the differential bacteria genera and differential metabolites. RESULTS:: showed that high-dose and low-dose SZD-JF groups had high content of MDA in intestinal tissue, low GSH content and SOD activity, short intestinal villi(P<0.05), low diversity and abundance of intestinal flora, variation in the intestinal flora structure, and low content of short chain fatty acids(P<0.05) compared with the normal group. Compared with high-dose and low-dose SZD-JF groups, high-dose and low-dose SZD groups displayed low content of MDA in intestinal tissue, high GSH content and SOD activity, recovery of the length of intestinal villi, increased abundance and diversity of intestinal flora, alleviation of dysbacteria, and recovery of the content of short chain fatty acids(P<0.05). According to the variation of intestinal flora and fecal metabolites after the addition of Jujubae Fructus, 6 differential bacterial genera(Lactobacillus, Butyricimonas, Clostridia＿UCG-014, Prevotella, Escherichia-Shigella, Alistipes),4 differential short chain fatty acids(such as acetic acid, propionic acid, butyric acid, valeric acid) and 18 differential metabolites(such as urolithin A, lithocholic acid, and creatinine) were screened out. Beneficial bacteria such as Lactobacillus were in positive correlation with butyric acid and urolithin A(P<0.05). The pathogenic bacteria such as Escherichia-Shigella were in negative correlation with propionic acid and urolithin A(P<0.05). In summary, SZD-JF caused obvious intestinal injury to normal rats, which could lead to intestinal flora disorder. The addition of Jujubae Fructus can alleviate the disorder and relieve the injury by regulating intestinal flora and the metabolites. This study discusses the effect of Jujubae Fructus in relieving the intestinal injury caused by SZD and the mechanism from the perspective of intestinal flora-host metabolism, which is expected to serve as a reference for clinical application of this prescription.

Mechanisms of copper transporter 1 gene in regulating radiation induced intestinal injury / 中华放射医学与防护杂志

Objective:To investigate the effects and mechanisms of copper transporter 1 (CTR1) in radiation induced intestinal injury in vitro. Methods:Human small intestinal epithelial cells (HIEC) were irradiated with 2, 4, 6, 8 Gy of X-rays and rat intestinal epithelial cells (IEC-6) were irradiated with 5, 10, 15, 20 Gy of X-rays. At 2, 4, 8, 24, and 48 h after irradiation, the expression of CTR1 was detected by Western blot assay. In some experiments, HIEC and IEC-6 cells were transfected with CTR1 shRNA and then exposed to X-rays. Copper levels were detected by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). The radiosensitivity of cells was verified by colonogenic assay, the cellular reactive oxygen species (ROS) level and DNA damage were detected to further explore the related mechanism. In addition, Western blot was applied to detect the expressions of antioxidants and cuproptosis associated proteins in enterocytes after silencing CTR1 or irradiation.Results:The expression of CTR1 was increased by X-ray irradiation in a dose-dependent manner ( t=3.53, 3.45, 6.37, 11.11, 11.13, P<0.05). CTR1 expression was successfully diminished by CTR1 shRNA adenovirus vectors. According to the survival curves, the enhancement ratios of the radiosensitivity of HIEC and IEC-6 cells with CTR1 knocking-down were 1.146 and 1.201, respectively. Radiation-induced copper accumulation was alleviated after CTR1 silencing in IEC-6 cells ( t=3.10, P<0.05). At 0.5 h after irradiation, the ROS production in the CTR1 knockdown group was significantly lower than that in the control group ( t=5.23, 2.96, P<0.05). At 1 h after irradiation, the protein expression of γ-H2AX in the CTR1 knockdown group was obviously lower than that in the control group ( t=7.50, 4.29, P<0.05). The expressions of Nrf2 and HO-1 were increased after irradiation, which could be further increased after CTR1 silencing. In addition, cuproptosis associated protein DLAT, LIAS and FDX1 were reduced post-irradiation, which were recovered after CTR1 silencing. Conclusions:The radioresistance of HIEC and IEC-6 cells was enhanced after CTR1 silencing, possibly through the intracellular ROS and cuproptosis pathway.

Analysis of whole genome expression profile for the effect of melatonin on radiation-induced intestinal injury in mice / 中华放射医学与防护杂志

Objective:To elucidate the change of whole genome expression profile for the effect of melatonin on radiation-induced intestinal injury in mice.Methods:C57BL/6J male mice were administrated with melatonin at 10 mg/kg body weight by intraperitoneal injection once a day for five consecutive days before abdominal irradiation with 14 Gy of γ-rays. Small intestines were harvested 3 d after radiation. GO annotation and KEGG pathway of the differential genes involved in small intestine were explored by DNA microarray analysis.Results:Compared with the control group, 584 differential genes were up-regulated and 538 differential genes were down-regulated for administration group pre-irradiation. The overlapping differential genes were selected from the irradiated mice and the administrated mice pre-irradiation. There were 324 up-regulated genes and 246 down-regulated genes unique to the administrated mice pre-irradiation. GO annotation analysis of the differential genes indicated that the top 15 significantly enriched biological processes for the administrated mice pre-irradiation mainly included autophagosome assembly (GO: 0000045), autophagosome organization (GO: 1905037) and regulation of acute inflammatory response (GO: 0002673). The genes ATG12, ATG16L2 and AMBRA1 were involved in autophagosome assembly and autophagosome organization. The genes C3, CPN1, CD55, CFP, CNR1, C1QA, C2 and CREB3L3 were involved in the regulation of acute inflammation response. KEGG pathway analysis of the differential genes involved indicated that the top 15 significantly enriched pathways for the administrated mice pre-irradiation mainly included O-glycan biosynthesis (hsa00512), glycosphingolipid biosynthesis (hsa00603), ECM-receptor interaction (hsa04512) and biosynthesis of unsaturated fatty acids (hsa01040). qRT-PCR verification showed that the expressions of ATG12 and ATG16L2 genes involved in autophagy for the administrated mice pre-irradiation increased significantly compared with the irradiated mice ( t=2.40, 4.35, P<0.05). Conclusions:The differential genes related with the biological process of autophagy, acute inflammatory response and the pathway of unsaturated fatty acid biosynthesis might be involved in the effect of melatonin on radiation-induced intestinal injury.

Effects of early enteral nutrition intervention on systemic inflammation and intestinal injury through NF-κB pathway in rats with acute pancreatitis / 中华内分泌外科杂志

Objective:To investigate the effects of early enteral nutrition intervention on systemic inflammation and intestinal injury in rats with acute pancreatitis and its mechanism.Method:Rat acute pancreatitis model was established. The rats were divided into sham surgery groups, model group, 12 h nutrition support group, 24 h nutrition support group, 48 h nutrition support group, and 48 h nutrition support group +PMA group according to the random number chart method, with 10 rats in each group. After laparotomy, the rats in sham operation group were closed after gently turning the pancreas. The sham operation group and model group were injected with the same amount of physiological salt. Nutritional support group for 12 h, nutritional support group for 24 h and nutritional support group for 48 h were given enteral nutrition support for 12, 24 and 48 h, respectively. Nutritional support group for 48 h +PMA group, intraperitoneal injection of 5 mg/kg NF-κB signaling pathway activator PMA was given after modeling, and nutritional support was given for 48 h. The contents of lipase, amylase and creatinine in serum of each group were detected by automatic biochemical analyzer. The serum levels of interleukin-6 (IL-6), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10) and D-lactic acid were detected by enzyme-linked immunosorbent assay (ELISA). The content of diamine oxidase (DAO) was detected by colorimetry. Hematoxylin-eosin (HE) staining was used to detect the pathological changes of intestinal mucosa. Western blot was used to detect the expression of NF-κB pathway-related proteins in pancreatic tissue of rats in each group.Results:(1) Lipase, amylase and creatinine in sham operation group, model group, 12 h nutrition support group, 24 h nutrition support group and 48 h nutrition support group were (4.37±0.61) vs (12.021±1.00) vs (8.77±0.62) vs (6.88±0.63) vs (5.20±0.41) U/ml, (1674.03±172.24) vs (4356.30±229.38) vs (3676.11±382.43) vs (2990.06±251.93) vs (1919.75±179.40) U/L, (32.12±3.37) vs (91.73±9.76) vs (72.38±6.83) vs (53.72±5.98) vs (41.82±4.00) U/L. Compared with sham operation group, the contents of serum lipase, amylase and creatinine in model group were significantly increased. Compared with model group, the contents of lipase, amylase and creatinine were significantly decreased after 12, 24 and 48 h of nutritional support, and were time-dependent ( P<0.05). (2) The levels of IL-6, IL-1β, TNF-α and IL-10 were (40.26±3.93) vs (123.34±13.19) pg/ml in sham operation group, model group, 12 h nutritional support group, 24 h nutritional support group and 48 h nutritional support group, respectively vs (108.97±12.70) vs (77.36±6.75) vs (49.18±4.97) pg/ml, (77.53±9.95) vs (316.36±23.76) vs (254.79±13.96) vs (177.92±17.20) vs (119.19±13.17) pg/ml, (62.94±5.39) vs (353.16±28.03) vs (275.87±22.11) vs (198.78±24.33) vs (94.60±9.41) pg/ml, (41.21±4.29) vs (6.92±1.01) vs (10.76±0.66) vs (21.24±1.64) vs (35.33±1.69) pg/ml. Compared with sham operation group, the contents of serum inflammatory cytokines IL-6, IL-1β and TNF-α in model group were significantly increased, while the content of IL-10 was significantly decreased. Compared with model group, the contents of IL-6, IL-1β and TNF-α were significantly decreased after 12, 24 and 48 h of nutritional support, while the contents of IL-10 were significantly increased in a time-dependent manner ( P<0.05). (3) The intestinal histopathological scores, DAO and D-lactic acid of sham operation group, model group, 12 h nutritional support group, 24 h nutritional support group and 48 h nutritional support group were (0.00±0.00) vs (4.20±0.60) vs (3.00±0.45) points, respectively vs (1.90±0.54) vs (1.30±0.64) points, (4.92±0.42) vs (14.95±1.20) vs (11.87±1.13) vs (9.02±0.53) vs (6.30±0.59) U/L, (2.39±0.22) vs (6.92±0.46) vs (5.21±0.28) vs (3.64±0.39) vs (2.95±0.15) nmol/ml. Compared with sham operation group, intestinal histopathological scores, DAO and D-lactic acid levels were significantly increased in model group. Compared with model group, intestinal histopathological scores, DAO and D-lactic acid levels were significantly decreased after 12, 24 and 48 h of nutritional support ( P<0.05). (4) The protein expressions of NF-κB p65 and p-IκBα were (0.23±0.03) vs (0.94±0.10) vs (0.75±0.06) vs (0.62±0.06) in sham operation group, model group, 12 h nutrition support group, 24 h nutrition support group and 48 h nutrition support group, respectively. vs (0.41±0.06), (1.06±0.12) vs (0.25±0.04) vs (0.47±0.03) vs (0.62±0.08) vs (0.85±0.08). Compared with sham operation group, NF-κB p65 protein level in model group was significantly increased, while p-IκBα protein level was significantly decreased. Compared with model group, the NF-κB p65 protein level was significantly decreased after 12, 24 and 48 h of nutritional support, while the P-iκBα protein was significantly increased ( P<0.05). (5) NF-κB p65, p-IκBα, IκBα, IL-6, IL-1β, TNF-α, IL-10, lipase, amylase and creatinine were (0.41±0.06) vs (0.82±0.06) in the 48 h group and the 48 h +PMA group, respectively. (0.85±0.08) vs (0.37±0.02), (1.05±0.11) vs (1.10±0.14), (49.18±4.97) vs (105.68±10.69) pg/ml, (119.19±13.17) vs (247.16±23.41) pg/ml, (94.60±9.41) vs (328.24±30.86) pg/ml, (5.20±0.41) vs (10.33±1.01) U/ml, (1919.75±179.40) vs (4023.40±334.56) U/L, (5.20±0.41) vs (10.33±1.01) U/ml, (41.82±4.00) U/L vs (81.33±7.96) U/L. Compared with the 48 h group, the expression level of NF-κB p65 protein, IL-6, IL-1β, TNF-α, lipase, amylase and creatinine in the 48 h +PMA group were significantly increased, while the expression level of P-iκBα protein and the content of IL-10 were significantly decreased ( P<0.05) . Conclusion:Early nutritional intervention can inhibit inflammatory response, reduce intestinal injury and control the development of acute pancreatitis by regulating NF-κB signaling pathway.

Progress in the prevention and treatment of traditional Chinese medicine based on the mechanism of intestinal injury of various chemotherapy / 中国临床药理学与治疗学

Intestinal injury is a common adverse reaction of clinical chemotherapy drugs, which limits the further application of chemotherapy drugs and causes serious physical and mental burden to patients. At present, the mechanism of chemotherapy-induced intestinal injury is complex, and traditional Chinese medicine has an excellent preventive effect. This article reviews the related mechanisms of intestinal flora imbalance, oxidative stress, inflammatory response, cell apoptosis, and immune damage caused by chemotherapy, and summarizes the role of traditional Chinese medicine in prevention and treatment of oxidative stress, inflammatory response, cell apoptosis, and immune damage.

Dahuang Mudantang Alleviates Intestinal Injury in Rat Model of Acute Pancreatitis by Regulating HMGB1/RAGE/NF-κB Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of Dahuang Mudantang in alleviating the intestinal injury in the rat model of acute pancreatitis via the high-mobility group box 1 （HMGB1）/receptor for advanced glycation endproduct （RAGE）/nuclear factor-κB （NF-κB） signaling pathway. MethodOne hundred and twenty SPF-grade Wistar rats received retrograde injection of 5% sodium taurocholate into the biliopancreatic duct for the modeling of intestinal injury in acute pancreatitis. The rats were randomized into blank， model， low-， medium-， and high-dose （3.5， 7， 14 g·kg-1， administrated by gavage） Dahuang Mudantang， and octreotide （1×10-5 g·kg-1， subcutaneous injection） groups （n=20）. The rats in blank and model groups received equal volume of distilled water by gavage. Drugs were administered 1 h before and every 12 h after modeling， and samples were collected 24 h after modeling. The general status of the rats was observed. The biochemical methods were employed to measure the levels of amylase （AMS） and C-reactive protein （CRP） in the serum. The enzyme-linked immunosorbent assay （ELISA） was employed to measure the levels of tumor necrosis factor （TNF）-α， interleukin （IL）-1β， and IL-6 in the colon tissue. The morphological changes of pancreatic and colon tissues were observed by hematoxylin-eosin （HE） staining. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot were employed to measure the expression levels of HMGB1， RAGE， inhibitor of NF-κB kinase （IKK）， and NF-κB suppressor protein α（IκBα）in the colon tissue. ResultThe rats in the model group showed poor general survival， writhing response， reduced frequency of defecation， and dry stool. The symptoms of rats in the model group were mitigated in each treatment group， and the high-dose Dahuang Mudantang showed the most significant effect. Compared with the normal group， the model group had elevated AMS and CRP levels （P<0.05）， which were lowered by Dahuang Mudantang （P<0.05）， especially that at the high dose （P<0.05）. Compared with the normal group， the modeling elevated that levels of TNF-α， IL-1β， and IL-6 （P<0.05）. Such elevations were lowered by Dahuang Mudantang （P<0.05）， and the high-dose group and the octreotide group showed better performance （P<0.05）. The modeling caused necrotic， congested， and destructed pancreatic and colonic tissues， which were ameliorated by the drugs， especially high-dose Dahuang Mudantang. Compared with the normal group， the modeling up-regulated the mRNA levels of HMGB1， RAGE， IKK， IκBα， and NF-κB （P<0.05）. Compared with the model group， Dahuang Mudantang and octreotide down-regulated the mRNA levels of HMGB1， RAGE， IKK， IκBα， and NF-κB （P<0.05）， and the high-dose Dahuang Mudantang demonstrated the best performance （P<0.05）. Western blot results showed a trend consistent with the results of Real-time PCR. ConclusionDahuang Mudantang can improved the general status， reduce inflammation， and alleviate histopathological changes in the pancreatic and colon tissues in the rat model of acute pancreatitis by inhibiting the HMGB1/RAGE/NF-κB signaling pathway.

Advances in research on small intestinal injuries caused by nonsteroidal anti-inflammatory drugs and its prevention and treatment / 中国药科大学学报

@#Nonsteroidal anti-inflammatory drugs (NSAIDs) are clinically used with common gastrointestinal adverse reactions, among which NSAIDs-induced small intestinal injuries (NSIs) are manifested in the appearance of jejunum and ileal mucosa erythema, erosion, ulcer, hemorrhage, intestinal wall perforation and obstruction et al..The pathological mechanisms of NSIs are complex, with a lack of effective prevention or treatment methods.This review summarizes the research progress of the pathological mechanisms of NSIs as well as the prevention and treatment of NSIs by misoprostol, mucosal protective agents, antibiotics and probiotics, traditional Chinese medicines and their active ingredients, nutritional supplements and other drugs in the past five years, in order to provide reference and basis for the research and development of new NSIs drugs.

Discovery of the radio-protecting effect of Ecliptae Herba, its constituents and targeting p53-mediated apoptosis in vitro and in vivo

Radiation protection drugs are often accompanied by toxicity, even amifostine, which has been the dominant radio-protecting drug for nearly 30 years. Furthermore, there is no therapeutic drug for radiation-induced intestinal injury (RIII). This paper intends to find a safe and effective radio-protecting ingredient from natural sources. The radio-protecting effect of Ecliptae Herba (EHE) was discovered preliminarily by antioxidant experiments and the mouse survival rate after 137Cs irradiation. EHE components and blood substances in vivo were identified through UPLC‒Q-TOF. The correlation network of "natural components in EHE-constituents migrating to blood-targets-pathways" was established to predict the active components and pathways. The binding force between potential active components and targets was studied by molecular docking, and the mechanism was further analyzed by Western blotting, cellular thermal shift assay (CETSA), and ChIP. Additionally, the expression levels of Lgr5, Axin2, Ki67, lysozyme, caspase-3, caspase-8,8-OHdG, and p53 in the small intestine of mice were detected. It was found for the first time that EHE is active in radiation protection and that luteolin is the material basis of this protection. Luteolin is a promising candidate for RⅢ. Luteolin can inhibit the p53 signaling pathway and regulate the BAX/BCL2 ratio in the process of apoptosis. Luteolin could also regulate the expression of multitarget proteins related to the same cell cycle.

Mechanism of the influence of CPT1 on the proliferation of rat intestinal epithelial cells IEC-6 after 60Co γ-ray irradiation / 中华放射医学与防护杂志

Objective:To investigate the changes of CPT1A and CPT1B protein expression in rat intestinal epithelial cells (IEC-6) after 60Co γ-ray irradiation, and the mechanism of the influence of carnitine palmitoyltransferase 1 (CPT1) on the proliferation of irradiated IEC-6 cells. Methods:IEC-6 cells were cultured in serum-normal medium or in serum-starved medium overnight, and pretreated with 20 μmol/L palmitic acid (PA) before irradiation with 0, 5, 10, and 15 Gy. At 24 h after irradiation, the cellular protein was collected for the measurement of CPT1A and CPT1B proteins by Western blot. The influences of ETO, an inhibitor of CPT1, on the survival and proliferation of irradiated IEC-6 cells were analyzed by colony formation assay and CCK-8 assay. The protein expressions and phosphorylation levels of the extracellular signal-regulated kinase (ERK1/2) and c-Jun N-terminal kinase (JNK) in 5 Gy irradiated IEC-6 cells pre-treated with ETO were analyzed by Western blot at 48 h after radiation.Results:When IEC-6 cells were cultured in serum-normal medium together with PA, the protein level of CPT1A was significantly increased after 15 Gy irradiation ( t=-2.82, P<0.05). When IEC-6 cells were cultured in serum-starved medium, the protein level of CPT1A was significantly increased at 5, 10, and 15 Gy ( t=-3.28, -8.72, -8.67, P<0.05). When IEC-6 cells were cultured in serum-starved medium together with PA, the protein levels of CPT1A were significantly increased at 5, 10 and 15 Gy ( t=-10.69, -7.02, -8.23, P<0.05), the protein levels of CPT1B were significantly increased at 10 and 15 Gy ( t=-3.73, -5.05, P<0.05). After irradiation, the survival and proliferation of IEC-6 cells in ETO group were significantly lower than those in control group ( t=5.46, 13.22, P<0.05), and the protein level of ERK1/2 and p-JNK in ETO group were significantly lower than those in control group ( t=4.01, 3.29, 10.68, 14.44, P<0.05). Conclusions:CPT1 promoted radiation-induced IEC-6 injury cells survival and proliferation by enhancing the expression level of ERK1/2 protein and the activity of JNK.

Research progress on tryptophan metabolism involved by gut microbiota in the diagnosis and treatment of acute radiation-induced intestinal injury / 中华放射肿瘤学杂志

Gut microbiota not only affects the activity of tryptophan metabolism rate limiting enzymes in intestinal cells, but also cooperatively produces a variety of catalytic enzymes, which directly affects the type and quantity of tryptophan metabolites in the intestine. Multiple tryptophan-associated indole compounds originating from the gut microbiome are significantly decreased in the peripheral blood of mice, and negatively correlated with radiation dose ranging from 2 to 10.4 Gy, which might be biomarkers for acute radiation-induced intestinal injury. Recent studies have reported that indole 3-propionic acid (IPA), indole-3-carboxaldehyde (I3A) and kynurenic acid (KYNA), which are tryptophan catabolites derived from gut microbiota, aryl hydrocarbon receptor, which is one of the receptors for tryptophan catabolites, and inhibition of indoleamine 2,3 dioxygenase-1, which is a main rate-limiting enzyme in intestinal tryptophan catabolism, can protect against radiation-induced intestinal toxicity. A more comprehensive understanding of the dynamics of tryptophan catabolites and their roles in acute radiation-induced intestinal injury is needed to deepen the understanding of the pathogenesis in radiation-induced intestinal injury and exploration of effective diagnostic and therapeutic approaches.

Preventive and therapeutic effects of Bateri-7 on radiation-induced intestinal injury in mice / 中华放射医学与防护杂志

Objective:To study the protective effect of Mongolian medicine Bateri-7 on radiation-induced intestinal injury in mice.Methods:C57BL/6J male mice were randomly divided into control group, irradiation group and irradiation plus drug administration group, with 10 or 15 mice in each group. For irradiation group, the mice were given a single dose of 12 Gy 60Co γ-rays with total body irradiation. For drug treatment, the mice were gavaged with Bateri-7 (530 mg/kg) 7 d before irradiation until 3 d after IR. At 6 h and 24 h after irradiation, the Tunel positive cells in intestine were detected immunohistochemically. At 3.5 d after irradiation, the structure of intestinal villi was observed by HE staining, and the BrdU and Ki67 positive cells were detected immunohistochemically. The expression levels of IL-6, TNF-α and Cxcl-5 were detected by qPCR. The FITC-dextran in peripheral blood was also determined. Results:The survival of irradiated mice was significantly increased by Bateri-7 ( χ2= 5.84, P < 0.05), but there was no significant difference in weight between two groups ( P > 0.05). The villi length of small intestine in the irradiation plus drug group was significantly longer than that in the irradiation group ( t = 20.24, P < 0.05), and there was no significant difference in the depth of intestinal crypt between two groups ( P > 0.05). At 6 and 24 h after irradiation, the number of Tunel positive cells in intestinal crypts in the irradiation plus drug group was significantly reduced in comparison with the irradiation group ( t = 3.52, 2.90, P < 0.05). At 3.5 d after irradiation, the level of FITC-dextran in serum and the expressions of IL-6, TNF-α and Cxcl-5 in small intestine of mice in the irradiation plus drug group were significantly lower than those in the irradiation group, respectively( t = 6.92, 7.01, 7.18, 13.16, P < 0.05). The number of BrdU and Ki67 positive cells in the crypt of mice in the irradiation plus drug group was higher than that of the irradiation group ( t = 3.91, 2.57, P < 0.05). Conclusions:Mongolian medicine Bateri-7 can effectively alleviate irradiation-induced intestinal injury of mice, which may have a good preventive and therapeutic effect on radiation enteritis.

Recombinant human ADAMTS13 ameliorates radiation-induced intestinal injury by regulating inflammation and oxidative stress / 中华放射医学与防护杂志

Objective:To study the protective effect of recombinant human ADAMTS13 (rhADAMTS13) on radiation-induced intestinal injury.Methods:Thirty C57BL/6 J mice were randomly divided into healthy control group, 12 Gy abdominal irradiation group (simple irradiation group) and rhADAMTS13 combined with 12 Gy irradiation group (combined group) with 6, 12 and 12 mice per group. The combined group was given 2.5 μg/kg rhADAMTS13 via tail vein injection 3 d before irradiation. 12 Gy X-ray abdominal irradiation was given to the simple irradiation group.The mice were executed at 1 and 3 d after irradiation, and plasma vWF, ADAMTS13 and CRP concentrations were measured by enzyme-linked immunosorbent assay (ELISA). Intestinal histopathological changes were observed by hematoxylin-eosin (HE) staining.Intestinal Ki67, TNF- α and MPO expression levels were detected by immunohistochemical staining.Plasma malondialdehyde (MDA), superoxide dismutase (SOD) activity and total antioxidant capacity (T-AOC) were measured by biochemical kits.Results:Compared with the healthy control group, the plasma vWF level was increased( t=6.20、12.29, P<0.05) and ADAMTS13 level was decreased( t=9.82、22.83, P<0.05)in mice at 1(1.38±0.11)and 3 (1.70±0.10)after irradiation.Compared with the simple irradiation group, the plasma vWF level was reduced ( t=2.93, 3.96, P<0.05) and ADAMTS13 level was increased ( t=5.09, 9.82, P<0.05) in mice at 1 (1.23±0.12) and 3 d (1.48±0.09) after irradiation in the combined group. Immunohistochemical staining showed that the number of Ki67 + cells in the crypt of the combined group increased significantly ( t=9.82, P<0.05) and the degree of TNF-α and MPO infiltration decreased significantly ( t=15.44, 14.33, P<0.05) compared with the simple irradiation group.In addition, rhADAMTS13 intervention significantly reduced plasma CRP ( t=5.02, 2.96, P<0.05) and MDA ( t=2.47, 2.55, P<0.05), but increased SOD activity ( t=2.64, 5.64, P<0.05) and T-AOC ( t=3.05, 5.07, P<0.05). Conclusions:rhADAMTS13 attenuates radiation-induced intestinal injury in mice by reducing the levels of inflammation and oxidative stress, providing a new strategy for the protection of radiation-induced intestinal injury.

Effect and mechanism of Rho kinase inhibitor on intestinal injury in septic rats / 中华危重病急救医学

Objective:To explore the effect of Rho kinase inhibitor on intestinal injury in septic rats and its possible mechanism.Methods:Thirty-two male Sprague-Dawley (SD) rats were randomly divided into sham operation group (Sham group), Rho kinase inhibitor Y-27632 control group (Y+Sham group), sepsis model group [cecal ligation and puncture (CLP) group] and Y-27632 pretreatment group (Y+CLP group), with 8 rats in each group. Rat sepsis model was reproduced by CLP. The rats in the Sham group and Y+Sham group were only separated and moved the cecum without ligation and perforation. The rats in the Y+Sham group and Y+CLP group were pretreated with intraperitoneal injection of Y-27632 solution 5 mg/kg 15 minutes before operation; the rats in the Sham group and CLP group were intraperitoneally injected with the same amount of phosphate buffered saline (PBS). Twenty-four hours after operation, the heart blood was collected and the serum diamine oxidase (DAO) content was determined by enzyme-linked immunosorbent assay (ELISA). Then the small intestine tissue was collected, the pathological changes of the intestinal tissue were observed under the light microscope after hematoxylin-eosin (HE) staining, and Chiu's score was performed. The positive expressions of Rho-related coiled-coil kinase 1 (ROCK1) and nuclear factor-κB (NF-κB) in intestinal tissue were detected by immunohistochemistry. ELISA was used to detect the levels of tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) in intestinal tissue homogenate.Results:The intestinal tissue structure of the Sham group and Y+Sham group was intact and the mucosa was arranged neatly. Compared with the Sham group, the intestinal mucosa of the CLP group was arranged disorderly, with a large number of inflammatory cells infiltration, and the Chiu's score was significantly increased (3.83±0.27 vs. 0.12±0.11, P < 0.05), indicating that those rats suffered from septic intestinal injury. Compared with the CLP group, the degree of necrosis of intestinal epithelial cells in the Y+CLP group was reduced, a small amount of inflammatory cells infiltration was seen, and the Chiu's score was significantly decreased (2.85±0.21 vs. 3.83±0.27, P < 0.05), indicating that Y-27632 pretreatment could alleviate intestinal injury in septic rats. Compared with the Sham group, the positive expressions of intestinal tissue ROCK1 and NF-κB, the contents of serum DAO and intestinal homogenate TNF-α in the CLP group were significantly increased [ROCK1 expression ( A value): 0.19 (0.18, 0.22) vs. 0.10 (0.09, 0.11), NF-κB expression ( A value): 0.40±0.02 vs. 0.15±0.01, DAO (ng/L): 287.81±23.31 vs. 144.92±17.72, TNF-α (ng/L): 101.08±5.62 vs. 74.81±5.56, all P < 0.05], the level of intestinal homogenate IL-10 was significantly decreased (μg/L: 55.16±5.20 vs. 95.95±7.53, P < 0.05). Compared with the CLP group, the positive expressions of intestinal tissue ROCK1, NF-κB, the contents of serum DAO and intestinal homogenate TNF-α in the Y+CLP group were significantly decreased [ROCK1 expression ( A value): 0.15 (0.13, 0.18) vs. 0.19 (0.18, 0.22), NF-κB expression ( A value): 0.28±0.01 vs. 0.40±0.02, DAO (ng/L): 243.34±19.76 vs. 287.81±23.31, TNF-α (ng/L): 90.41±8.79 vs. 101.08±5.62, all P < 0.05], while the level of intestinal homogenate IL-10 was significantly increased (μg/L: 66.15±5.74 vs. 55.16±5.20, P < 0.05), indicating that the protective effect of Y-27632 pretreatment on sepsis intestinal injury rats might be related to the regulation of RhoA/ROCK1/NF-κB signaling pathway. Conclusion:Rho kinase inhibitors can reduce intestinal injury in septic rats, and the mechanism may be related to inhibiting RhoA/ROCK1/NF-κB signaling pathway and reducing intestinal inflammation in septic rats.

Effect of MiR-320 on Intestinal Injury in Rats With Acute Pancreatitis by Regulating JAK2/STAT3 and NF-κB Signaling Pathways / 胃肠病学

Background: Expression of microRNA⁃320 (miR⁃320) is down regulated in acute pancreatitis, and the mechanism of its effect on acute pancreatitis is still unclear. Aims: To investigate the effect of miR⁃320 on intestinal injury in rats with acute pancreatitis and its mechanism. Methods: Rats were randomly divided into sham operation group, model group, miR⁃ 320 agonist group (agomir miR ⁃ 320 group), miR ⁃ 320 agonist control group (agomir NC group), JAK2 inhibitor group (AG490 group), and NF⁃κB pathway inhibitor group (PDTC group). The rat model of acute pancreatitis was established by retrograde injection of 5% sodium taurocholate to the bile duct. The automatic biochemical analyzer was used to detect serum levels of amylase and lipase; ELISA assay was used to detect serum levels of TNF⁃α and IL⁃1β; HE staining was used to observe the pathological changes of rat pancreas and ileum; TUNEL staining was used to observe cell apoptosis in rat ileum; real⁃time fluorescent quantitative PCR (RT⁃qPCR) was used to detect the expression of miR⁃320 in ileum tissue; Western blotting method was used to detect the expressions of JAK2/STAT3 and NF⁃κB signaling pathway related proteins in ileum. Results: Compared with sham operation group, the pancreas and ileum were severely injured in model group, and the pathological score and ileum cell apoptosis were significantly increased (P<0.05), serum levels of amylase, lipase, TNF⁃ α, and IL⁃1β were significantly increased (P<0.05), the expression of miR⁃320 in ileum tissue was significantly decreased (P<0.05), the ratios of p⁃JAK2/JAK2, p⁃STAT3/STAT3, p⁃p65/p65, and p⁃IκBα/IκBα in ileum tissue were significantly increased (P<0.05). Compared with model group, the pathological damages of pancreas and ileum in agomir miR ⁃ 320 group, AG490 group and PDTC group were reduced, and the pathological score and ileum cell apoptosis were significantly decreased (P<0.05), serum levels of amylase, lipase, TNF ⁃ α, and IL ⁃ 1β were significantly decreased (P<0.05), the expression of miR⁃320 in ileum tissue was significantly increased (P<0.05), the ratios of p⁃JAK2/JAK2, p⁃STAT3/STAT3, p⁃ p65/p65, and p⁃IκBα/IκBα in ileum tissue were significantly decreased (P<0.05). Conclusions: MiR⁃320 can improve the intestinal injury in rats with acute pancreatitis by inhibiting the activation of JAK2/STAT3 and NF⁃κB signaling pathways.

The key role of gut-liver axis in pyrrolizidine alkaloid-induced hepatotoxicity and enterotoxicity

Pyrrolizidine alkaloids (PAs) are the most common phytotoxins with documented human hepatotoxicity. PAs require metabolic activation by cytochromes P450 to generate toxic intermediates which bind to proteins and form protein adducts, thereby causing cytotoxicity. This study investigated the role of the gut-liver axis in PA intoxication and the underlying mechanisms. We exposed mice to retrorsine (RTS), a representative PA, and for the first time found RTS-induced intestinal epithelium damage and disruption to intestinal barrier function. Using mice with tissue-selective ablation of P450 activity, we found that hepatic P450s, but not intestinal P450s, were essential for PA bioactivation. Besides, in RTS-exposed, bile duct-cannulated rats, we found the liver-derived reactive PA metabolites were transported by bile into the intestine to exert enterotoxicity. The impact of gut-derived pathogenic factors in RTS-induced hepatotoxicity was further studied in mice with dextran sulfate sodium (DSS)-induced chronic colitis. DSS treatment increased the hepatic endotoxin level and depleted hepatic reduced glutathione, thereby suppressing the PA detoxification pathway. Compared to RTS-exposed normal mice, the colitic mice displayed more severe RTS-induced hepatic vasculature damage, fibrosis, and steatosis. Overall, our findings provide the first mode-of-action evidence of PA-induced enterotoxicity and highlight the importance of gut barrier function in PA-induced liver injury.

Safety analysis of definitive surgery for chronic radiation intestinal injury / 中华胃肠外科杂志

Objective: To investigate the safety of definitive surgery for chronic radiation intestinal injury. Methods: A descriptive case series study was performed. Clinical data of 105 patients who were diagnosed as chronic radiation intestinal injury, had complete data and received definitive surgery (the radiation-induced intestinal segment and digestive tract reconstruction) at Department of Gastrointestinal Surgery of Beijing Tsinghua Changgung Hospital from June 2016 to May 2020 were retrospectively analyzed. There were 30 males (28.6%) and 75 females (71.4%) with the median age of 58 years (P25, P75: 52, 64 years). Patients who had tumor recurrence or refused surgical treatment were excluded. According to the preoperative evaluation and clinical manifestations, to select the resection range. Outcome parameters: (1) preoperative evaluation (nutrition risk assessment and status of obstruction or fistula); (2) clinical manifestations and treatment strategies; (3) details of surgical parameters; (4) postoperative complications, and Clavien-Dindo classification III to V was defined as main moderate-severe complication. Results: (1) Preoperative evaluation: Eighty-eight patients (83.8%) developed symptoms of chronic radiation intestinal injury more than 1 year after the end of radiotherapy. Ninety-eight patients (93.3%) had preoperative NRS-2002 score ≥3, 74 patients (70.5%) received preoperative parenteral nutritional support, and the median time of nutritional support was 10.5 (7.0, 16.0) days. Sixteen patients (15.2%) received small intestinal decompression tube implantation due to severe obstruction. (2) Clinical manifestations and treatment strategies: Among 105 patients, 87 (82.9%) presented with obstruction and received definitive resection of the radiation-induced intestinal segment plus one-stage digestive tract reconstruction; 18 (17.1%) presented with intestinal fistula and all of them received definitive resection of the radiation-induced intestinal segment, intestinal fistula plus one-stage digestive tract reconstruction. Among above 18 patients with fistula, 3 patients with ileorectal stump fistula received pedicled pelvic closure of greater omentum at the same time; 4 patients had ileal vesical fistula, of whom 2 patients received cystectomy and bladder repair due to preoperative nephrostomy decompression, and the other 2 patients received transection of the small intestine proximal and distal to the fistula and anastomosis of the intestinal loop without fistula resection, intestinal fistula or bladder fistula repair. (3) The details of surgical parameters: Median operative time and intraoperative blood loss was 230 (180, 300) minutes and 50 (20, 50) ml respectively. Ninety-two patients (92/105, 87.6%) underwent ileocolonic anastomosis, and anastomosis on the hepatic flexure or splenic flexure colon were performed in 88 (83.8%) and 4 (3.8%) patients respectively. Ileoileal anastomosis was performed in 13 patients (12.4%). The anastomotic site of 92 patients (87.6%) was strictly located in the contralateral quadrant of the radiation field, and the anastomotic site of 13 patients (12.4%) was far from the radiation field. Nine patients (8.6%) had more than one anastomosis, 5 patients (4.8%) had less than 180 cm of residual small intestine, 7 patients (6.7%) underwent retrograde intestinal permutation, 4 patients (3.8%) underwent abdominal wall reconstruction surgery due to abdominal wall defects, and 87 patients (82.9%) had severe abdominal pelvic adhesions (grade 3-4 adhesions). Intraoperative complications occurred in 3 patients (2.9%), which were found in time and handled properly. The median postoperative hospital stay was 13.0 (12.0, 24.5) days, and all the patients had resumed oral feeding upon discharge. (4) Postoperative complications: Fourteen patients (13.3%) had 18 major complications (grade III to V). The incidence of postoperative anastomotic leakage was 5.7% (6/105), and the incidence of anastomotic leakage for ileocolon anastomosis and ileoileal anastomosis was 2.2% (2/92) and 4/13, respectively (χ(2)=17.29, P<0.001). The incidence of postoperative anastomotic leakage of intestinal fistula and intestinal obstruction was 3/18 and 3.4% (3/87), respectively (χ(2)=4.84, P=0.028). The mortality at 30 days after operation was 1.0% (1/105), after abdominal infection and septic shock caused by postoperative anastomotic leakage resulting in multiple organ failure. Conclusion: For chronic radiation intestinal injury patients with obstruction or fistula, definitive surgical treatment is feasible and safe with acceptable major complications.

Role of intestinal stem cells in prevention of radiation-induced intestinal injury / 中华放射医学与防护杂志

At present, there is no effective treatment method of radiation-induced intestinal injury. Clinically, only symptomatic remission therapy can be used. Intestinal stem cells (ISCs) have strong renewal ability, which is an important part of intestinal structure regeneration. To promote the recovery of the number and function of ISCs has been a research hotspot in the treatment of radiation gastrointestinal syndrome. This article introduces the important role of ISCs in the protection of radiation-induced intestinal injury and its clinical application significance from the classification and regulatory mechanism of ISCs, and drugs that regulate the damage, apoptosis and repair of stem cells, so as to provide reference for the future research on the role of intestinal stem cells in radiation protection.

Correlation of intestinal fatty acid binding protein and diamine oxidase with intestinal injury in strangulated bowel obstruction mice / 中华内分泌外科杂志

Objective:To investigate the correlation of intestinal fatty acid binding protein and diamine oxidase with intestinal injury in strangulated bowel obstruction mice.Methods:160 SD rats were divided into 5 groups by random number table: group A ( n=32) : normal control group; group B ( n=32) : sham operation group; group C ( n=32) : simple intestinal obstruction group; The strangulated intestinal obstruction group was divided into group D ( n=32) : acute superior mesenteric artery ischemia group and group E ( n=32) : acute mesenteric arterial and venous ischemia. Except group A, other groups were given operation for modeling. Venous blood and small intestinal segment of group A was collected after anaesthesia, and venous blood and small intestinal segment of other groups were collected after modeling for 4 h. Serum samples were collected from venous blood, and intestinal fluid samples were collected by soaking the small intestinal segments. The intestinal segments were observed and the intestinal injury was evaluated. The levels of intestinal fatty acid binding protein (I-FABP) and activity of diamine oxidase (DAO) in serum and intestinal fluid were detected. Pearson correlation analysis was used to analyze the correlation between intestinal injury and the serum and intestinal fluid levels of I-FABP and activity of DAO, respectively. Results:The intestinal damage scores in group B, C, D and E were higher than that in group A, the intestinal damage scores in groups C, D and E were higher than that in group B, the intestinal damage scores in groups D and E were higher than that in group C, and the intestinal damage score in group E was higher than that in group D ( P<0.05) . The serum I-FABP level and DAO activity in group C, D and E were higher than those in group A and B, and the serum I-FABP level and DAO activity in group D and E were higher than those in group C ( P<0.05) . The level of I-FABP and DAO activity in intestinal fluid in group C, D and E were higher than those in groups A and B, and the level of I-FABP and DAO activity in intestinal fluid in group D and E were higher than those in group C ( P<0.05) . There were positive correlations between intestinal injury and the serum and intestinal fluid levels of I-FABP and activity of DAO, respectively ( r=0.972, P<0.001; r=0.899, P<0.001; r=0.961, P<0.001; r=0.828, P<0.001) . Conclusions:Intestinal injury of strangulated bowel obstruction mice is related to the intestinal ischmia. There are increases of serum and intestinal fluid levels of I-FABP and activity of DAO in strangulated bowel obstruction mice, which are closely related to the degree of intestinal injury.