The miR-9-5p/CXCL11 pathway is a key target of hydrogen sulfide-mediated inhibition of neuroinflammation in hypoxic ischemic brain injury

We previously showed that hydrogen sulfide(H2S)has a neuroprotective effect in the context of hypoxic ischemic brain injury in neonatal mice.However,the precise mechanism underlying the role of H2S in this situation remains unclear.In this study,we used a neonatal mouse model of hypoxic ischemic brain injury and a lipopolysaccharide-stimulated BV2 cell model and found that treatment with L-cysteine,a H2S precursor,attenuated the cerebral infarction and cerebral atrophy induced by hypoxia and ischemia and increased the expression of miR-9-5p and cystathionineβsynthase(a major H2S synthetase in the brain)in the prefrontal cortex.We also found that an miR-9-5p inhibitor blocked the expression of cystathionineβsynthase in the prefrontal cortex in mice with brain injury caused by hypoxia and ischemia.Furthermore,miR-9-5p overexpression increased cystathionine-β-synthase and H2S expression in the injured prefrontal cortex of mice with hypoxic ischemic brain injury.L-cysteine decreased the expression of CXCL11,an miR-9-5p target gene,in the prefrontal cortex of the mouse model and in lipopolysaccharide-stimulated BV-2 cells and increased the levels of proinflammatory cytokines BNIP3,FSTL1,SOCS2 and SOCS5,while treatment with an miR-9-5p inhibitor reversed these changes.These findings suggest that H2S can reduce neuroinflammation in a neonatal mouse model of hypoxic ischemic brain injury through regulating the miR-9-5p/CXCL11 axis and restoringβ-synthase expression,thereby playing a role in reducing neuroinflammation in hypoxic ischemic brain injury.

Metabolite acetyl-L-carnitine participates in Bifidobacterium animalis F1-7 to ameliorate atherosclerotic inflammation by downregulating theTLR4/NF-κB pathway

This study aimed to explore the effect of Bifidobacterium animalis F1-7 on the improvement of atherosclerotic inflammation.Arteriosclerosis model ApoE^(-/-)mice were orally administered with B.animalis F1-7 for 12 weeks.The probiotic intervention reduced the plaque areas in aorta and the accumulation of macrophages,and downregulated the expression of toll-like receptor 4(TLR4)/nuclear factorκB(NF-κB)pathway to reduce the levels of inflammatory factors.The widely-targeted metabolomics analysis showed that acetyl-L-carnitine(ALC)in the intestine of atherosclerotic mice was significantly increased after B.animalis F1-7 intervention.Correlation analysis proved that ALC was associated with atherosclerotic inflammatory response.By using oxidized low density lipoprotein induced macrophage foam cells,we further verified that ALC could reduce lipid accumulation and inflammatory response in foam cells by downregulating the TLR4/NF-κB pathway.Finally,our results revealed that B.animalis F1-7 upregulated the metabolite ALC to downregulate the inflammatory responses,leading to the reduction of plaque accumulation of atherosclerosis.

Biological function of miRNA-145-5p in angiotensin II induced renal inflammation

Objective:Chronic kidney disease(CKD)is a progressive disorder characterized by intricate structural and functional alterations in the kidneys,attributable to diverse causative factors.Notably,the therapeutic promise of miR-145-5p in addressing renal pathologies has been discerned.This investigation seeks to elucidate the functional role of miR-145-5p in injured kidneys by subjecting human glomerular mesangial cells(HGMCs)to stimulation with Angiotensin II(AngII).Materials and Methods:Cellular viability and the levels of inflammatory mediators were evaluated utilizing Cell Counting Kit-8(CCK-8),quantitative real-time polymerase chain reaction(qRT-PCR),and western blot methodologies,both in the presence of AngII incubation and in scenarios of miR-145p overexpression and downregulation.Furthermore,the cell cycle dynamics were elucidated through Fluorescence-activated Cell Sorting(FACS)analysis.Results:AngII incubation induced an upregulation of miR-145-5p and inflammatory factors including Intercellular Adhesion Molecule 1(ICAM-1),Interleukin 6(IL-6),Interleukin 8(IL-8),and Interleukin 1β(IL-1β).Additionally,it elevated the expression of Cyclin A2,Cyclin D1,and the G2/M cell cycle ratio.Conversely,inhibition of miR-145-5p heightened the levels of inflammatory factors and cell cycle regulators induced by AngII incubation.Reduced expression of miR-145-5p correlated with a downregulation of Interleukin 10(IL-10)expression,concurrently promoting HGMC proliferation under AngII stimulation.Moreover,ectopic miR-145-5p expression demonstrated a reduction in inflammatory factors,cell cyclin regulators,G2/M cell cycle ratio,and overall proliferation.Conclusion:MiR-145-5p exhibited inhibitory effects on the inflammatory response and proliferation induced by Angiotensin II in HGMCs,showcasing its potential as a therapeutic avenue for the treatment of kidney injury.

Safety and in Vivo Anti-Inflammatory Activity of Ethanolic Extract of Ficus umbellata (Vahl.) Leaves

Toxicity is the totality of adverse effects, which can be functional and morphological lesions in a living organism, caused by a substance introduced in relatively high single doses or in small, repeated doses. The aim of this study was to assess the OECD-recommended acute oral toxicity and anti-inflammatory activity of ethanolic extract of Ficus umbellata leaves. Animals were given a single oral dose of 1000, 3000 and 5000 mg/Kg body weight (BW) of the extract. For the anti-inflammatory activity test, rats were given the ethanolic extract of F. umbellata leaves at doses of 100, 300 and 500 mg/Kg or aspirin® at a concentration of 100 mg/Kg PC orally, one hour before injection of 0.05 ml of 1% formalin under the plantar fascia of the rat’s right hind paw. Paw volume measurements were taken one, two and three hours after formalin injection, using an electronic caliper. After 14 days of observation, no deaths were observed in treated rats. The LD50 of ethanolic extract of Ficus umbellata leaf powder is greater than 5000 mg/Kg body weight. This extract has no significant effects on hematological parameters and on the main markers of nephrotoxicity and hepatotoxicity for a single dose of less than 5000 mg/Kg PC. It reduces formalin-induced edema. Evaluation of the percentage inhibition showed that the extract had greater anti-inflammatory activity at 3 hours after the start of the experiment. However, better inhibition of inflammatory oedema of the paw of rats treated with 500 mg/Kg was observed at 5 hours after the start of the experiment, with a percentage inhibition of 69.23 ± 1.02, compared with the reference group treated with aspirin® 100 mg/Kg, which showed an inhibition of 63.50 ± 0.98. These results show that F. umbellata leaves possess anti-inflammatory activity, which would justify their use in traditional African medicine to prevent or treat inflammation.

Tamanu Oil in Acne Management: Potential Anti-Inflammatory and Wound-Healing Properties for Scar Reduction

Tamanu oil, derived from the nuts of Calophyllum inophyllum, has gained increasing attention for its potential in acne management due to its purported anti-inflammatory and wound-healing properties. This analysis evaluates the efficacy of tamanu oil in acne treatment with a specific focus on its impact on inflammation and scar reduction. The novelty of this research lies in its comprehensive analysis of tamanu oil’s dual mechanism of action: reducing acne-related inflammation and promoting the healing of acne scars. Clinical trials and laboratory analyses were conducted to assess the oil’s effectiveness in diminishing erythema, swelling, and post-acne scarring compared to conventional treatments. Preliminary findings demonstrate that tamanu oil significantly reduces inflammation and accelerates wound healing, potentially offering a promising adjunct or alternative to standard acne therapies. Future research should aim to optimize formulation and application protocols, long-term effects, and comparative therapeutic efficacy with other anti-inflammatory agents. Tamanu oil offers a novel and effective approach to acne management, with potential advantages that go beyond inflammation reduction to include enhanced scar reduction, making it a subject that warrants further investigation.

Prophylactic Anti-inflammation Inhibits Cigarette Smoke-induced Emphysema in Guinea Pigs

In this study, the effect of prophylactic anti inflammation on the development of smoke induced emphysema was investigated. Young male guinea pigs aged 1.5 - 2 months (weighing 198.3±26.9 g) were randomly divided into 4 groups: group A (cigarette smoke exposure only), group B (cigarette smoke exposure plus pentoxifylline rich (PTX, 10 mg/d) forage feeding), group C (cigarette smoke exposure plus intermittent cortical steroid injection (Triamcinolone acetonide, 3 mg, im, every three weeks) and control group (group D: animals with sham smoke exposure, raised under the same conditions). Animals in group A, B and C were exposed to smoke of cigarettes for 1 to 1.5 h twice a day, 5 days a week. All animals were killed at the 16th week and followed by morphometrical analysis of the midsagittal sectioned lung slices. Smoke exposure of 16 weeks resulted in visible emphysematous development in Group A but not in Group B and C. It was evidenced by the indicator of air space size, mean linear intercept (L m): 120.6±16.0 μm in Group A; 89.8±9.2 μm in Group B and 102.4±17.7 μm in Group C. The average L m in either group B or group C was shorter than that in Group A (ANOVA and Newman Keuls test, F=8.80, P =0.0002) but comparable to that (94.8±13.2 μm) in group D ( P >0.05). It is concluded that long term prophylactic anti inflammation inhibits pulmonary emphysema induced by cigarette smoking in the guinea pigs.

Diabetic retinopathy:Role of inflammation and potential therapies for anti-inflammation

Diabetic retinopathy is a leading cause of blindness among working-age adults.Despite many years of research,treatment options for diabetic retinopathy remain limited and with adverse effects.Discovery of new molecular entities with adequate clinical activity for diabetic retinopathy remains one of the key research priorities in ophthalmology.This review is focused on the therapeutic effects of cannabidiol(CBD),a nonpsychoactive native cannabinoid,as an emerging and novel therapeutic modality in ophthalmology based on systematic studies in animal models of inflammatory retinal diseases including diabetic retinopathy-a retinal disease associated with vascular-neuroinflammation.Special emphasis is placed on novel mechanisms which may shed light on the pharmacological activity asso c iated with CBD preclinically.These include a selfdefence system against inflammation and neurodegeneration mediated by inhibition of equilibrat ive nucleoside transporter and activation of adenosine receptor by treatment with CBD.

Neuroprotective mechanisms of rutin for spinal cord injury through anti-oxidation and anti-inflammation and inhibition of p38 mitogen activated protein kinase pathway

Rutin has anti-inflammatory, antioxidant, anti-viral, anti-tumor and immune regulatory effects. However, the neuroprotective effects of rutin in spinal cord injury are unknown. The p38 mitogen activated protein kinase （p38 MAPK） pathway is the most important member of the MAPK family that controls inflammation. We assumed that the mechanism of rutin in the repair of spinal cord injury is associated with the inhibition of p38 MAPK pathway. Allen’s method was used to establish a rat model of spinal cord injury. The rat model was intraperitoneally injected with rutin （30 mg/kg） for 3 days. After treatment with rutin, Basso, Beattie and Bresnahan locomotor function scores increased. Water content, tumor necrosis factor alpha, interleukin 1 beta, and interleukin 6 levels, p38 MAPK protein expression and caspase-3 and -9 activities in T8–9 spinal cord decreased. Oxidative stress related markers superoxide dismutase and glutathione peroxidase levels increased in peripheral blood. Rutin exerts neuroprotective effect through anti-oxidation, anti-inflammation, anti-apoptosis and inhibition of p38 MAPK pathway.

Anti-inflammation and anti-fibrosis actions of resveratrol on experimental pneumoconiosis

OBJECTIVE To investigate the therapeutic effects and related signaling pathways involved in the actions of resveratrol on experimental pneumoconiosis in vivo and in vitro.METHODS The pneumoconiosis animal model was induced by exposing male SD rats to 15mg·m-3 silica aerosol in an inhalation chamber system for 6h·d-1,5d·week-1 for up to 8 weeks.The vehicle or resveratrol(10or 20mg·kg-1)was preventively or remedially administered to the rats during or after the 4-or 8-week silica exposure(SE)period,respectively.After 4-,8-,and up to 14-week treatment,in vivo near-infrared fluorescence imaging analysis and histological analysis were performed to evaluate the pathological changes in rat lung.Inflammatory cytokines level in bronchoalveolar lavage fluid(BALF)was measured by ELISA testing,and the deposition of fibrotic collagen proteins in lung parenchyma was determined by western blotting and immunohistochemistry analysis.Microarray analysis was performed to screen the signaling pathways involved in the actions of resveratrol on pneumoconiosis in vitro models.Anti-inflammation action and signaling of resveratrol was evaluated on silica-stimulated rat alveolar macrophage,which is one of the crucial effector cells for silica-induced inflammatory response;anti-fibrosis action and signaling of resveratrol was evaluated on TGF-β-induced human lung fibroblast,which acts as a promoter in the later fibrotic process of pneumoconiosis.RESULTS Silica aerosol exposure significantly increased macrophage infiltration and matrix metalloproteinases activity in lung tissue concomitant with the increased levels of inflammatory mediators in BALF.Preventive treatment with resveratrol(20mg·kg-1·d-1)reversed all these biochemical indices as well as histopathological alterations induced by silica exposure.Post-SE resveratrol treatment mildly reduced silica-induced inflammatory response in rat lung with no statistical significance.In vitro study revealed that resveratrol could inhibit alveolar macrophage cell death and decrease the levels of IL-1β and TNF-αinduced by silica particle exposure to cultured alveolar macrophages.Resveratrol was further shown to inhibit the nuclear transition of NF-κB and formation of cleaved caspase-1.Encouragingly,resveratrol preventively attenuated the lung fibrosis,evidenced by less fibrotic nodules formation and collagen proteins expression.No significant improvement on lung fibrosis was observed with post-SE resveratrol treatment.Invitrostudy further demonstrated that resveratrol suppressed TGF-β-induced lung fibroblast proliferation and collagen deposition,concomitant with the depressed activity of TGF-β/Smad signaling in lung fibroblast.CONCLUSION Resveratrol shows the anti-inflammation and anti-fibrosis actions on experimental pneumoconiosis in vivo and in vitro models.The depression of NF-κB,NALP3-inflammasome,and TGF-β/Smad signaling pathways may be involved in the anti-inflammation and anti-fibrosis actions of resveratrol,respectively.Resveratrol could be a potential therapeutic agent for the intervention of pneumoconiosis.

18种中药多糖与Siglec-9的亲和力测试及其抗炎作用研究

目的比较18种中药多糖与唾液酸特异性Ig样凝集素-9(sialic-acid-binding immunoglobulin-like lectin,Siglec-9)的亲和力以及抗炎能力,探讨中药多糖发挥抗炎作用的新机制。方法选择18种不同药性的中药用水提醇沉法提取多糖,苯酚硫酸法检测多糖含量,高效液相色谱法检测多糖的单糖组成,红外光谱法检测中药多糖结构。通过竞争性ELISA法检测多糖与Siglec-9的亲和力,并研究中药多糖对LPS诱导巨噬细胞炎症反应以及R848诱导中性粒细胞NETosis的影响,利用髓系细胞特异性敲除Siglec-E(Lyz2-cre:Siglec-E flox/flox)小鼠来源的原代巨噬细胞和腹腔中性粒细胞进一步明确多糖结合Siglec-9产生抗炎作用的机制。结果中药多糖的糖含量均在55%以上,主要由甘露糖、葡萄糖、半乳糖、阿拉伯糖、葡萄糖醛酸等组成,但单糖构成比各不相同。红外光谱检测4种代表性中药多糖均有典型的多糖吸收峰,柴胡和蒲公英多糖具有葡萄糖醛酸结构。枸杞和黄芪多糖与Siglec-9的亲和力,对LPS诱导的巨噬细胞炎症反应和R848诱导的中性粒细胞NETosis反应无显著抑制作用。柴胡和蒲公英多糖与Siglec-9亲和力较强(P<0.01),能够显著抑制上述炎症反应,但其抗炎作用在髓系细胞特异性敲除Siglec-E基因小鼠来源的巨噬细胞和中性粒细胞中消失。结论中药多糖与Siglec-9的亲和力有潜力成为研究中药抗炎作用的新的现代药理学机制。

丝裂霉素C-全氟溴辛烷脂质体纳米药物对人翼状胬肉成纤维细胞治疗安全性及效果评价

目的制备一种以脂质体为载体,以液态的全氟溴辛烷(PFOB)为内核,丝裂霉素C(MMC)搭载于脂质体壳的纳米药物PFOB@Lip-MMC,研究其对人翼状胬肉成纤维细胞(HPF)增殖的抑制作用。方法采用薄膜分散-水化超声法制备PFOB@Lip-MMC,并检测其物理、化学性质。通过CCK-8、Cam-PI细胞活/死染色、流式细胞术等方法检测不同浓度PFOB@Lip-MMC对HPF活性的影响。在激光共聚焦显微镜下采用DiI荧光标记的PFOB@Lip-MMC观察纳米药物对于HPF的药物渗透性。建立HPF炎症细胞模型后,按照实验要求分为对照组(加入无菌PBS溶液)、PFOB@Lip组(加入PFOB@Lip)、MMC组(加入MMC)、PFOB@Lip-MMC组(加入PFOB@Lip-MMC)、正常组(加入新鲜培养基),共孵育24 h后,流式细胞仪检测炎症细胞的凋亡率以及从PCR角度分析细胞中白细胞介素(IL)-1β、前裂腺素E_(2)(PGE_(2))、肿瘤坏死因子(TNF)-α及血管内皮生长因子(VEGF)基因表达水平。结果PFOB@Lip-MMC平均粒径和Zeta电位分别为(103.45±2.17)nm和(27.34±1.03)mV,其包封率和载药率分别为(72.85±3.28)%和(34.27±2.04)%。体外24 h眼表环境下载药纳米微球缓释MMC可达(78.34±2.92)%。PFOB@Lip-MMC的生物安全性较MMC明显提高。DiI荧光标记的PFOB@Lip-MMC与炎症化HPF共同孵育2 h后,DiI荧光标记弥漫性分布于炎症化HPF中。PFOB@Lip-MMC组炎症化HPF细胞凋亡率[(77.23±4.93)%]明显高于MMC组[(51.62±3.28)%];PCR检查结果显示,其余各组IL-1β、PGE 2、TNF-α及VEGF基因基因转录水平与对照组和PFOB@Lip组相比均明显下降,其中PFOB@Lip-MMC组降低最明显,差异均有统计学意义(均为P<0.05)。结论本研究成功合成了一种新型的抗HPF增殖的纳米药物PFOB@Lip-MMC,且细胞毒性较原药明显降低,具有良好的生物相容性和抗炎效果,为降低翼状胬肉术后复发率提供了一种新的治疗思路。

白刺花叶总生物碱通过抑制MAPK/NF-κB信号通路减轻脂多糖诱导的RAW264.7细胞炎症反应

目的探讨白刺花叶总生物碱(Sophora davidii Hance leaves total alkaloids,SDLTAs)的体外抗炎作用及可能的分子机制。方法采用脂多糖(LPS)诱导RAW264.7细胞炎症模型,给予不同浓度(50、100、200μg/mL)SDLTAs,Griess法检测SDLTAs对细胞NO表达的影响;ELISA法检测SDLTAs对细胞因子IL-6、TNF-α和IL-1β表达的影响;RT-qPCR法检测细胞中iNOS、NF-κB p65和IκBαmRNA的表达;Western blotting法检测细胞中p-p38、p-p65和p-JNK以及细胞核内NF-κB p65的蛋白表达。结果SDLTAs显著抑制LPS诱导的RAW264.7细胞炎症反应。SDLTAs极显著降低细胞中NO、IL-6、TNF-α和IL-1β的分泌(P<0.01),极显著下调细胞中iNOS、NF-κB p65和IκBαmRNA的表达(P<0.01),极显著降低细胞中p-p38、p-p65和p-JNK以及细胞核内NF-κB p65的蛋白表达(P<0.01)。结论SDLTAs可通过调节MAPK/NF-κB信号通路发挥抗炎作用。

玉米黄素减轻脂多糖诱导的3T3-L1脂肪细胞慢性炎症

目的:探究一种天然类胡萝卜素——玉米黄素(ZEA),基于NF-κB信号通路减轻脂多糖(LPS)诱导的3T3-L1脂肪细胞慢性炎症并发挥抗炎的作用机制。方法:以LPS刺激成熟的3T3-L1脂肪细胞建立炎症模型,用不同浓度的ZEA(0,5,10,15μmol/L)进行干预,检测脂肪细胞炎症相关因子的含量,并通过实时荧光定量PCR(qPCR)检测其mRNA表达水平,用蛋白免疫印迹法检测信号通路因子的蛋白表达水平。结果:与LPS组相比,ZEA干预显著抑制炎症细胞模型中促炎因子IL-1β、IL-6、TNF-α和MCP-1分泌(P<0.001),同时促进抗炎因子IL-10和ADPN产生(P<0.001)。此外,qPCR结果显示ZEA能有效抑制促炎因子,同时恢复抗炎因子的mRNA表达水平(P<0.001)。蛋白免疫印迹结果表明,ZEA能明显降低IκB激酶催化亚基IKKα和IKKβ的蛋白和磷酸化表达(P<0.001),增强抑制蛋白IκBα与核转录因子NF-κB p65的结合,最终抑制NF-κB p65进行核内转录。结论:ZEA可通过抑制NF-κB信号通路激活及调控炎性因子的分泌,改善肥胖引起的慢性炎症。

High fat diet dysregulates microRNA-17-5p and triggers retinal inflammation:Role of endoplasmic-reticulum-stress

AIM To elucidate how high diet-induced endoplasmic reticulum-stress upregulates thioredoxin interacting protein expression in Müller cells leading to retinal inflammation. METHODS Male C57Bl/J mice were fed either normal diet or 60% high fat diet for 4-8 wk. During the 4 wk study, mice received phenyl-butyric acid(PBA); endoplasmic reticulum-stress inhibitor; for 2 wk. Insulin resistance was assessed by oral glucose tolerance. Effects of palmitate-bovine serum albumin(BSA)(400 μmol/L) were examined in retinal Müller glial cell line and primary Müller cells isolated from wild type and thioredoxin interacting protein knock-out mice. Expression of thioredoxin interacting protein, endoplasmic reticulum-stress markers, mi R-17-5p m RNA, as well as nucleotide-binding oligomerization domain-like receptor protein(NLRP3) and IL1β protein was determined.RESULTS High fat diet for 8 wk induced obesity and insulin resistance evident by increases in body weight and impaired glucose tolerance. By performing quantitative real-time polymerase chain reaction, we found that high fat diet triggered the expression of retinal endoplasmic reticulum-stress markers(P < 0.05). These effects were associated with increased thioredoxin interacting protein and decreased mi R-17-5p expression, whichwere restored by inhibiting endoplasmic reticulumstress with PBA(P < 0.05). In vitro, palmitate-BSA triggered endoplasmic reticulum-stress markers, which was accompanied with reduced mi R-17-5p and induced thioredoxin interacting protein m RNA in retinal Müller glial cell line(P < 0.05). Palmitate upregulated NLRP3 and IL1β expression in primary Müller cells isolated from wild type. However, using primary Müller cells isolated from thioredoxin interacting protein knock-out mice abolished palmitate-mediated increase in NLRP3 and IL1β.CONCLUSION Our work suggests that targeting endoplasmic reticulumstress or thioredoxin interacting protein are potential therapeutic strategies for early intervention of obesityinduced retinal inflammation.

Potential role of chitinase 3-like-1 in inflammation-associated carcinogenic changes of epithelial cells

The family of mammalian chitinases includes members both with and without glycohydrolase enzymatic activity against chitin, a polymer of N-acetylglucosamine. Chitin is the structural component of fungi, crustaceans, insects and parasitic nematodes, but is completely absent in mammals. Exposure to antigens containing chitin- or chitin-like structures sometimes induces strong T helper type-I responses in mammals, which may be associated with the induction of mammalian chitinases. Chitinase 3-like-1 （CHI3L1）, a member of the mammalian chitinase family, is induced specifically during the course of inflammation in such disorders as inflammatory bowel disease, hepatitis and asthma. In addition, CHI3L1 is expressed and secreted by several types of solid tumors including glioblastoma, colon cancer, breast cancer and malignant melanoma. Although the exact function of CHI3L1 in inflammation and cancer is still largely unknown, CHI3L1 plays a pivotal role in exacerbating the inflammatory processes and in promoting angiogenesis and remodeling of the extracellular matrix. CHI3L1 may be highly involved in the chronic engagement of inflammation which potentiates development of epithelial tumorigenesis presumably by activating the mitogen-activated protein kinase and the protein kinase B signaling pathways. Anti-CHI3L1 antibodies or pan-chitinase inhibitors may have the potential to suppress CHI3Ll-mediated chronic inflammation and the subsequent carcinogenic change in epithelial cells.

基于血清抗-HBc定量建立慢性HBV感染者显著肝组织病理学改变的无创诊断模型

目的建立慢性HBV感染者显著肝组织病理学改变的诊断模型,并评估模型的诊断价值。方法选取2011年12月—2017年12月在上海市公共卫生临床中心肝胆内科住院并进行肝活检且未经抗病毒治疗的457例慢性HBV感染者,收集患者的肝活检病理结果及常规实验室指标,并进行抗-HBc定量检测。依据Scheuer方法进行炎症分级(G)及纤维化分期(S),分为显著与非显著肝坏死性炎症、肝纤维化和肝损伤组。根据单因素分析和多因素logistic回归分析构建预测显著肝坏死性炎症、显著肝纤维化和显著肝损伤的数学模型,并与FIB-4、GPR、APRI、RPR利用ROC曲线分析评估模型的预测性能,根据ROC曲线下面积(AUC)比较诊断价值。结果457例患者中显著肝坏死性炎症(G≥2)178例,非显著肝坏死性炎症(G<2)279例,显著肝纤维化(S≥2)248例,非显著肝纤维化(S<2)209例,显著肝损伤(G≥2或/和S≥2)264例,非显著肝坏死性炎症(G<2和S<2)193例。根据单因素分析和多因素logistic回归分析结果,分别建立由抗-HBc、AST、PLT、TTR等指标组成的预测显著肝坏死性炎症的数学模型M-SHN,由抗-HBc、PLT、ChE、TTR、性别等指标组成的预测显著肝纤维化的数学模型M-SHF,及由抗-HBc、PLT、TTR、性别等指标组成的预测显著肝损伤的数学模型M-SHI,并通过ROC曲线分析各模型的预测价值,M-SHN模型的AUC为0.826(95%CI:0.788~0.860),M-SHF模型的AUC为0.776(95%CI:0.735~0.814),M-SHI模型的AUC为0.789(95%CI:0.748~0.825)。结论基于患者常规实验室指标及血清抗-HBc定量,建立了M-SHN、M-SHF、M-SHI模型,对于显著肝坏死性炎症、显著肝纤维化、显著肝损伤有较为可靠的预测价值,可帮助临床评估患者是否需要进行抗病毒治疗。

地塞米松磷酸钠纳米制剂对IL-1β诱导大鼠原代软骨细胞炎症的影响

目的:探讨地塞米松(Dex)磷酸钠纳米制剂(Nano-Dex)对白细胞介素(IL)-1β诱导的大鼠原代软骨细胞炎症的影响。方法:利用透射电镜观察Nano-Dex纳米粒的形态,马尔文粒径仪检测Nano-Dex的粒径和电位。培养SD大鼠软骨细胞,将细胞分为空白组、IL-1β组、IL-1β+Dex组和IL-1β+Nano-Dex组。用实时荧光定量PCR(RT-qPCR)法检测软骨细胞基质金属蛋白酶(MMP)13、MMP3、IL-1β、IL-6、Ⅱ型胶原α1(Col2a1)和蛋白聚糖(ACAN)基因表达水平,免疫荧光染色观察MMP13的表达,western blotting法检测软骨细胞MMP13、Col2a1、IL-6、IL-1β、P65和Caspase 3蛋白表达水平。结果:Nano-Dex形状圆润,大小均一,直径约在90 nm,带正电荷。与空白组比较,IL-1β组软骨修复基因Col2a1、ACAN表达下调,而炎症基因IL-1β、IL-6、MMP3、MMP13表达上调(均P<0.05);与IL-1β组比较,Nano-Dex+IL-1β组Col2a1基因及蛋白表达上调,ACAN基因表达上调,IL-1β、IL-6、MMP3、MMP13基因表达下调,MMP13、P65、IL-6、IL-1β和Caspase 3蛋白表达均下调(均P<0.05)。免疫荧光结果显示,IL-1β+Nano-Dex组MMP13的荧光强度明显低于IL-1β组(P<0.05)。结论:Nano-Dex能有效抑制IL-1β诱导的软骨细胞炎症。

Anti-inflammatory and Immunomodulatory Effects of Marine n-3 Polyunsaturated Fatty Acids on Human Health and Diseases

The pharmaceutical effects of n-3 polyunsaturated fatty acids(n-3 PUFAs) as dietary nutrients on human health and diseases have gained much attention and are investigated for decades. Docosahexaenoic acid(DHA), eicosapentaenoic acid(EPA) and docosapentaenoic acid(DPA) are the three major n-3 PUFAs enriched in marine organisms, such as fish, shrimp, algae, and so on. It has been well known that n-3 PUFAs, especially DHA and EPA, are beneficial in reducing the risk of cardiovascular and cerebrovascular diseases. Accumulating evidence suggests that n-3 PUFAs might cure inflammatory diseases through several mechanisms, such as plasma membrane remodeling of lymphocytes, down-regulating pro-inflammatory cytokines, and alternating adhesion molecule expressions. Several molecular targets of n-3 PUFAs on immune-regulation have also been identified, such as GPR120(FFA4), protein kinase C(PKC), and PPAR-γ. However, it remains inconclusive if dietary n-3 PUFAs function the same both in vitro and in vivo based on cohort studies. This review will focus on the molecular targets and mechanisms of anti-inflammatory and immunomodulatory effects of n-3 PUFAs on human health and diseases, such as obesity, tumor, diabetes, and autoimmune diseases.

竹节参总皂苷缓解CCl_(4)诱导的大鼠急性肝损伤:基于调控PI3K/Akt/NF-κB信号通路

目的探讨土家族药用植物竹节参提取物总皂苷对CCl_(4)致急性肝损伤的保护作用及潜在的药理学机制。方法将6周龄SPF级雄性SD大鼠随机分为正常组、模型组、联苯双酯组(100 mg/kg)、竹节参总皂苷低、中、高(50、100、200 mg/kg)剂量组,各组8只,除空白组外,其余各组采用CCl_(4)诱导大鼠急性肝损伤模型,处理组于造模中给予药物灌胃干预。比较各组大鼠的血清谷草转氨酶(AST)、谷丙转氨酶(ALT)、总胆红素(TBil)和碱性磷酸酶(ALP)水平;HE染色观察肝组织病理学改变;免疫组化检测肝脏组织PI3K/Akt/NF-κB信号通路相关分子的表达;酶联免疫法测定肝脏组织总超氧化物歧化酶(T-SOD)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)水平;蛋白免疫印迹法检测肝脏组织PI3K-Akt和SIRT6-NF-κB通路相关蛋白表达情况。结果网络药理学分析显示,竹节参总皂苷对急性肝损伤的有治疗作用,其关键的通路为PI3K/Akt等信号通路。血清学和酶联免疫学实验结果显示,与正常组相比,模型组大鼠血清和肝组织的AST、ALT、ALP、TBil和MDA明显增高(P<0.01),T-SOD和GSHPx水平显著降低(P<0.01);与模型组相比,各治疗组ALT、AST、ALP、TBIL和MDA水平显著降低(P<0.01),T-SOD和GSH-Px水平显著升高(P<0.01)。免疫组化结果显示,正常组大鼠肝组织细胞内未见p-NF-κB表达阳性,与正常组相比,模型组中p-NF-κB阳性表达则明显增加,阳性细胞数增多(P<0.01);与模型组比较,各治疗组阳性表达明显减少(P<0.01)。免疫印迹结果显示,相对于正常组,模型组PI3K、p-Akt蛋白的表达水平下降,p-NF-κB、TNF-α和IL-6蛋白的表达水平升高(P<0.05);与模型组相比,各治疗组PI3K、p-Akt和SIRT6蛋白的表达水平显著升高,p-NF-κB p65、TNF-α和IL-6的蛋白的表达水平显著降低(P<0.05)。结论竹节参总皂苷可以通过调节PI3K/Akt和NF-κB通路,有效减缓CCl_(4)诱导的大鼠急性肝损伤,发挥其抗炎、抗氧化应激和保护肝损伤的作用。

5-羟基-6,7-二甲氧基黄酮抑制流感病毒诱导A549细胞炎症反应和铁死亡的作用及机制

目的 研究四方蒿中提取出的5-羟基-6,7-二甲氧基黄酮(5-HDF)对H1N1流感病毒引起的肺损伤的保护作用及潜在机制。方法 使用H1N1流感病毒感染A549细胞制作流感病毒感染模型,检测5-HDF的细胞毒性及其对感染病毒后的炎症和铁死亡相关指标及相关信号通路蛋白的影响。采用乙醇回流提取和硅胶色谱法从四方蒿中提取分离获得5-HDF,并采用NMR和MS鉴定其结构。采用MTT法检测不同浓度的5-HDF对A549细胞的毒性;H1N1流感病毒以0.1的感染复数感染A549细胞;采用流式细胞术检测5-HDF对感染H1N1流感病毒后A549细胞中TRAIL和IL-8表达水平的影响;Western blotting检测phospho-p38 MAPK(Thr180/Tyr182)、phospho-NF-κB p65(Ser536)、cleaved caspase3、cleaved PARP、SLC7A11、GPX4等炎症、细胞凋亡和铁死亡相关蛋白表达的水平。通过鼻腔接种50μL半数致死量(LD50)的H1N1流感病毒液复制小鼠H1N1流感病毒感染模型,以体质量变化率、肺解剖结果、肺组织病理形态变化为指标,考察5-HDF(30 mg/kg、60 mg/kg)体内抗H1N1流感病毒的作用。结果 MTT结果显示5-HDF在0~200μg/mL对A549细胞没有明显的细胞毒性(P>0.05)。流式细胞术和Western blotting结果显示,5-HDF能抑制H1N1流感病毒感染的A549细胞中phospho-NF-κB p65和phospho-p38 MAPK的活化,降低促炎因子IL-8的表达,且5-HDF能上调SLC7A11和GPX4等抗铁死亡相关蛋白表达的水平,并抑制凋亡标志物cleaved caspase3和cleaved PARP及凋亡因子TRAIL的表达(P<0.05)。5-HDF灌胃给药7 d,可提高H1N1流感病毒感染导致的小鼠体质量降低,降低因感染H1N1流感病毒而异常升高的肺指数,并减轻其肺组织病变程度(P<0.05)。结论 本研究推测5-HDF具有一定的抗小鼠H1N1流感病毒感染作用,可能通过增加SLC7A11和GPX4的表达并抑制phospho-NF-κB p65和phospho-p38 MAPK的活化,降低cleaved caspase3和cleaved PARP的表达,来减弱流感病毒H1N1感染A549细胞所引起的铁死亡、炎症反应和细胞凋亡。