黄花菜甲醇提取物对糖脂损伤的干涉作用

该研究以秀丽隐杆线虫(Caenorhabditis elegans,C.elegans)作为模式生物研究了黄花菜甲醇提取物对糖脂损伤的干涉作用。该研究利用UPLC-QQQ-MS/MS对黄花菜甲醇提取物中的成分进行了分析,并以野生型N2秀丽线虫为基础,以100μmol/mL蔗糖诱导糖损伤模型,以100μg/mL硬脂酸诱导脂损伤模型,对三个模型分别喂食0、400、800、1200μg/mL浓度的黄花菜甲醇提取物,检测黄花菜甲醇提取物对秀丽线虫产卵量、身长、寿命表型的干涉作用。结果表明,黄花菜甲醇提取物中的主要成分为芦丁,含量为5721.11 ng/mg。对普通N2秀丽线虫,喂食800μg/mL浓度黄花菜甲醇提取物的线虫比空白组线虫的总子代数和身长分别提高24.76%和7.73%;对糖损伤秀丽线虫,喂食800μg/mL浓度黄花菜甲醇提取物的线虫比空白组线虫的寿命和总子代数分别提高24.47%和3.40%;对于脂损伤秀丽线虫,喂食1200μg/mL浓度黄花菜甲醇提取物的线虫比空白组线虫的寿命、身长和总子代数分别提高30.04%、10.36%和97.32%。因此,中高浓度的黄花菜甲醇提取物对秀丽线虫的糖脂损伤有一定的干涉作用。

单唾液酸四己糖神经节苷脂对急性颅脑损伤应用价值的临床观察

目的 :探讨单唾液酸四己糖神经节苷脂 (GM 1)在急性颅脑损伤治疗中的应用。方法 :对 5 2例急性颅脑损伤患者 ,伤后及早静脉滴注GM110 0mg·d-1,2周后改为 2 0mg肌内注射 ,bid ,共 10d。结果 :(1)治疗组 :8～ 15分患者和 3～ 7分患者治疗前、后伤情相比较差异具有显著性 ,P <0 .0 5 ;(2 )治疗组恢复良好和中残的病例是总数的 82 .6 %,对照组为 6 4.3%,两组比较差异具有显著性 ,P <0 .0 5。结论 :GM 1具有较好的催醒作用 ,对提高患者的生存质量、降低死亡率、促进脑功能恢复和改善预后具有较好疗效。

利用秀丽线虫糖/脂损伤模型快速鉴定4种天然产物的修复功效

高糖、高脂饮食会导致各种慢性代谢疾病。以野生型秀丽线虫N2为研究对象,通过分别喂食100 mmol/L蔗糖和100μg/mL硬脂酸,构建糖/脂损伤秀丽线虫模型。利用此模型分析日常膳食中的4种天然活性成分——胡椒碱、贝萼皂苷元、芍药色素-3-O-葡萄糖苷、天竺葵素-3-O-芸香糖苷对糖/脂损伤线虫的修复作用。结果:1)对糖损伤线虫,500μg/mL胡椒碱和1 000μg/mL天竺葵素-3-O-芸香糖苷有较好的修复作用,产卵总量分别提高25%和4.6%,线虫第4天的身长分别提高了16.84%和0.98%,寿命分别延长了8.1%和3.19%;1 000μg/mL芍药色素-3-O-葡萄糖苷对其生殖能力和生长发育有较好的修复作用,产卵总量提高了55.81%,而寿命缩短了22.57%;贝萼皂苷元具有显著的破坏作用。2)对脂损伤线虫,500μg/mL胡椒碱、500μg/mL贝萼皂苷元和500μg/mL芍药色素-3-O-葡萄糖苷均有一定修复作用,子代总量分别提高18.41%,13.41%和29.17%,线虫第3天的身长分别提高6.59%,3.19%和10.98%,寿命分别延长15.4%,2.84%和10.46%;天竺葵素-3-O-芸香糖苷对其生殖能力和生长发育有较好的修复作用,而不影响寿命。实验结果表明:4种天然产物对糖/脂损伤均有一定修复作用。此模型可作为快速鉴定天然产物对糖脂损伤修复作用的一种简单、可行的技术手段。

Poly-lactic acid and agarose gelatin play an active role in the recovery of spinal cord injury

Objective To investigate the role of poly-lactic acid and agarose gelatin in promoting the functional recovery of the injured spinal cord. Methods Poly-lactic acid （PLA） or agarose was embedded in the space between two stumps of the hemisectioned spinal cord. Immunohistochemistry was used to show astroglia proliferation and the infiltration of RhoA-positive cells. Locomotor activity recovery was evaluated by testing the function of hindlimbs. Results Astroglias and RhoA labeled non-neuronal cells accumulated in the area adjacent to the implant, while the number of RhoA-posirive cells was decreased dramatically in the absence of implant. Animals implanted with agarose gelatin recovered more quickly than those with PLA, concomitant with a higher survival rate of the neurons. Conclusion Both PLA and agarose gelatin benefited the recovery of spinal cord after injury by providing a scaffold for astroglia processes. Modulation of the rigidity, pore size and inner structure of PLA and agarose gelatin might make these biodegradable materials more effective in the regeneration of the central nervous system （CNS）.

Adiponectin deficiency exacerbates lipopolysaccharide/ D-galactosamine-induced liver injury in mice

AIM： To examine the effects of adiponectin on the functions of Kupffer cells, key modulators of lipopolysaccharide （LPS） -induced liver injury.METHODS： D-galactosamine （GAIN） and LPS were injected intraperitoneally into adiponectin-/- mice and wild type mice. Kupffer cells, isolated from Sprague-Dawley rats, were preincubated with or without adiponectin, and then treated with LPS.RESULTS： In knockout mice, GalN/LPS injection significantly lowered the survival rate, significantly raised the plasma levels of alanine transaminase and tumor necrosis factor-α （TNF-α） and significantly reduced IL-10 levels compared with wild type mice. TNF-α gene expression in the liver was which higher and those of IL-10 were lower in knockout mice than in wild type mice. In cultured adiponectin-pre-treated Kupffer cells, LPS significantly lowered TNF-α levels and raised IL-10 levels in the culture media and their respective gene expression levels, compared with Kupffer cells without adiponectinpre-treatment.CONCLUSION： Adiponectin supresses TNF-α production and induces IL-10 production by Kupffer cells in response to LPS stimulation, and a lack of adiponectin enhances LPS-induced liver injury.

Lipopolysaccharide Challenge Induces Long Pentraxin 3 Expression in Mice Independently from Acute Lung Injury

Objective To determine whether the onset of acute lung injury （ALl） induces the up-regulation of pentraxin 3 （PTX3） expression in mice and whether PTX3 concentration in the biofluid can help recognizing sepsis-induced ALI. Methods Wild-type C57BL/6 mice （12-14 weeks old） were randomly divided into 3 groups. Mice in the group 1 （n=12） and group 2 （n=12） were instilled with lipopolysaccharide via intratracheal or intraperitoneal routes, respectively. Mice in the group 3 （n=8） were taken as blank controls. Pulmonary morphological and functional alterations were measured to determine the presence of experimental ALl. PTX3 expression in the lung was quantified at both protein and mRNA levels. PTX3 protein concentration in blood and bronchoalveolar lavage fluid was measured to evaluate its ability to diagnose sepsis-induced ALI by computing area under receiver operator characteristic curve （AUROCC）. Results ALl was commonly confirmed in the group 1 but never in the other groups. PTX3 expression was up-regulated indiscriminately among lipopolysaccharide-challenged mice. PTX3 protein concentration in the biofluid was unable to diagnose sepsis-induced ALl evidenced by its small AUROCC. PTX3 concentration in bronchoalveolar lavage fluid did not correlate with that in serum. Conclusions Lipopolysaccharide challenges induced PTX3 expression in mice regardless of the presence ofALI. PTX3 may act as an indicator of inflammatory response instead of organ injury per se.

Immunomodulatory effect of pachymaran on cyclosporine A(CsA)-induced lung injury in mice

Objective To investigate the immunomodulatory effect of pachymaran on cyclosporine A(CsA)-induced lung injury in mice.Methods(i) Fifty male BALB/c mice were randomly divided into five groups(10 mice in each group): normal control(NC) group, 30, 45, and 60 mg/kg CsA groups, and lipopolysaccharide(LPS) group. Except for the NC group, other groups underwent CsA modeling. The NC group was treated with phosphate-buffered saline(PBS), the LPS group with 10 mg/kg LPS eight hours before mice euthanized, and the 30, 45, and 60 mg/kg CsA groups with corresponding doses of CsA for seven consecutive days. After treatment, the body and organ mass of each group were weighed, and the lung, thymus, and spleen indexes were calculated. Hematoxylin-Eosin(HE) staining was performed to observe histopathological changes in the lungs of the mice. The protein expression levels of interleukin(IL)-2 and IL-1β in the blood were detected using enzyme-linked immunosorbent assay(ELISA), and those of surfactant protein D(SP-D), IL-2, and IL-6 in lung tissues were detected by immunohistochemistry(IHC). The mRNA expression levels of SP-D, IL-1β, IL-6, and myeloperoxidase(MPO) in the lung tissues were detected by quantitative reverse transcriptase-polymerase chain reaction(q RT-PCR).(ii) Another 60 BALB/c mice were divided into six groups(10 mice in each group) : NC group,model control(MC) group, 50, 100, and 200 mg/kg pachymaran groups, and polyinosinicpolycytidylic acid [poly(I:C)] group. Except for the NC group, other groups underwent45 mg/kg CsA modeling. The NC and MC groups were treated with distilled water, the pachymaran groups with corresponding doses pachymaran, and the poly(I:C) group with 0.1 mg/kg poly(I:C) for seven days.The mice were euthanized to obtain tissues and serum for detection.Detection methods were identical to those described in(i) above.Results(i) CsA(30 mg/kg) increased the lung index of mice(P < 0.001), and decreased the spleen index(P < 0.01), thymus index(P < 0.05), and the serum level of IL-2(P < 0.05). CsA(45 mg/kg) decreased the spleen, thymus indexes, and the serum level of IL-2(P < 0.01) in mice, and increased the serum level of IL-1β(P < 0.05) and the protein level of lung SP-D(P <0.001). CsA(60 mg/kg) increased the lung index of mice(P < 0.01), the serum level of IL-1β(P < 0.05), the protein level of lung SP-D(P < 0.01), and the mRNA levels of lung MPO and SP-D( P < 0.05), and decreased the thymus index of mice(P < 0.01). HE staining showed that 30, 45, and60 mg/kg CsA, and LPS caused pathological changes in the lung tissue of mice.(ii) After pachymaran intervention in MC mice, the spleen and thymus indexes(P < 0.05) were increased in the 100 and 200 mg/kg pachymaran groups, and the lung index was decreased(P < 0.05).Moreover, 50 mg/kg pachymaran increased the thymus index(P < 0.05) and decreased the lung index(P < 0.01) in MC group. Pachymaran(50, 100, and 200 mg/kg) improved lung tissue injury, reduced the serum level of IL-1β(P < 0.001), and the mRNA levels of MPO and SPD in lung tissues(P < 0.05) of mice. Pachymaran(100 mg/kg) increased the protein level of lung IL-2(P < 0.01), decreased the protein level of lung SP-D(P < 0.01), and the mRNA level of IL-1β(P < 0.001) in the lung tissues of mice. Pachymaran(200 mg/kg) increased the serum level of IL-2(P < 0.01) and lung IL-6 of mice(P < 0.05). Pachymaran(50 and 200 mg/kg) increased the mRNA level of IL-6 in the lung tissues of mice(P < 0.05).Conclusion While the immune function of mice was suppressed by CsA, the lung tissue was also damaged. Pachymaran can improve the immunosuppression induced by CsA and improve the lung tissue injury in immunosuppressed mice.

Serial analysis of gene expression in mice with lipopoly-saccharide-induced acute lung injury

Objective: To monitor the systemic gene expression profile in a murine model of lipopolysaccharide-induced acute lung injury. Methods: Acute lung injury was induced by intratracheal injection of lipopolysaccharide in 3 mice. Another 3 normal mice receiving same volume of normal saline were taken as the controls. The comprehensive gene expression profile was monitored by the recently modified long serial analysis of gene expression. Results: A total of 24 670 tags representing 12 168 transcripts in the control mice and 26 378 tags representing 13 397 transcripts in the mice with lung injury were identified respectively. There were 11 transcripts increasing and 7 transcripts decreasing more than 10 folds in the lipopolysaccharide-treated mice. The most overexpressed genes in the mice with lung injury included serum amyloid A3, metallothionein 2, lipocalin 2, cyclin-dependent kinase inhibitor 1A, lactate dehydrogenase 1, melatonin receptor, S100 calcium-binding protein A9, natriuretic peptide precursor, etc. Mitogen activated protein kinase 3, serum albumin, complement component 1 inhibitor, and ATP synthase were underexpressed in the lung injury mice. Conclusions: Serial analysis of gene expression provides a molecular characteristic of acute lung injury.

Esculin alleviates acute kidney injury and inflammation induced by LPS in mice and its possible mechanism

Acute kidney injury(AKI)is a common clinical serious illness.Esculin(ES)is a coumarin compound of traditional Chinese medicine Cortex Fraxini.Our previous study has found that ES protects against inflammation and renal damage in diabetic rats.In the present study,we aimed to investigate the effects and the possible mechanism of ES against lipopolysaccharides(LPS)-induced AKI in mice.Renal morphology was observed by H&E staining.Renal function was evaluated by blood urea nitrogen(BUN)level and creatinine content in serum.Inflammatory factor levels were measured by ELISA assay.The inflammatory proteins were analyzed by RT-PCR and Western blotting analysis.The results showed that ES alleviated LPS-induced pathological injury and renal dysfunction,and decreased BUN level and creatinine content in serum.In addition,ES significantly reduced the release of pro-inflammatory factors,including IL-1β,IL-6 and TNF-α,chemokine MCP-1 and cell adhesion molecule ICAM-1.Furthermore,the expressions of inflammatory pathway proteins P2 X7,HMGB1,TLR4 and MyD88 both at the mRNA and protein levels were all down-regulated by ES in the kidney tissue of LPS-challenged mice.These results suggested ES protected against LPS-induced AKI through inhibiting P2 X7 expression and HMGB1/TLR4 inflammatory pathway.

Lipopolysaccharide/Toll-like receptor 4 signaling pathway involved Qingdu decoction treating severe liver injury merging with endotoxemia

OBJECTIVE: To investigate the protective effects and underlying mechanism of Qingdu decoction(QDD) on experimental rats with severe liver injury induced by thioacetamide(TAA).METHODS: A total of 40 Wistar rats were randomly divided into normal group(n = 10) and experimental group(n = 30). Rats were administrated the same content of saline in normal group. The rats inthe experimental group were pretreated with TAA at dose of 12 mg/kg lasting 8 weeks, and from 9th week to 12 th week, with TAA at concentration of 36mg/kg. During the 9th week to 12 th week period,the rats were randomly divided into three subgroups(n = 10 each) simultaneously based on the treatment categories: model group, lactulose(LA,3.5 m L/kg) group and QDD(5.95 g/kg) group, orally once per day respectively. At the 12 th week, the content of serum alanine transaminase(ALT), aspartate transaminase(AST), total bilirubin(TBIL), endotoxin(ET) and tumor necrosis factor α(TNF-α) was detected by automatic biochemical analyzer. The plasma prothrombin time(PT), prothrombin time-international normalized ratio(PTR) and prothrombin time activity(PTA) were measured by automatic coagulation analyzer. The level of lipopolysaccharide(LPS)-binding protein(LBP), cluster differentiation 14(CD14) and Toll-like receptor 4(TLR4) expressions was measured by both western blot(WB) and real-time polymerase chain reaction(real-time PCR).RESULTS: Compared with the model group, hepatic morphology in the QDD group was improved under light microscope and transmission electron microscope; at the same time, the contents of serum ALT, AST, TBIL, ET and TNF-α, and level of LBP, CD14 and TLR4 expressions in liver tissues were significantly decreased compared with the model group(P < 0.05), while PTA in the QDD group was enhanced(P < 0.05).CONCLUSION: QDD has the functional effect on improving the injured liver through inhibiting the LPS/TLR4 signaling pathway thus decreasing the level of the inflammatory medicators.