丹红注射液对动脉粥样硬化兔主动脉血红素氧合酶-1表达的影响研究

目的:观察丹红注射液对动脉粥样硬化兔主动脉斑块内血红素氧合酶-1表达的影响,探讨抗动脉粥样硬化的作用机制。方法:选择遗传性高脂血症兔36只,将其随机分为对照组、高脂模型组、丹红注射液组三组(n=12)。对照组及高脂模型组分别以普通饲料与高脂饲料喂养,丹红注射液组予以高脂饲料并丹红注射液[15ml/(kg·d)]腹腔内注射,持续饲养12周。采用免疫组织化学法及Western blot法测定三组兔主动脉中血红素氧合酶-1的表达,并进行统计学分析。结果:三组兔主动血管平滑肌与动脉粥样硬化斑块中均明显可见血红素氧合酶-1的表达。丹红注射液组兔的主动脉壁中血红素氧合酶-1表达低于高脂模型组,较对照组显著增高(P<0.05)。结论:丹红注射液可上调红素氧合酶-1在主动脉的表达,进而减轻动脉粥样硬化的发展。

4-1BB signaling activates glucose and fatty acid metabolism to enhance CD8^(+) T cell proliferation

4-1BB(CD137)is a strong enhancer of the proliferation of CD8^(+)T cells.Since these cells require increased production of energy and biomass to support their proliferation,we hypothesized that 4-1BB signaling activated glucose and fatty acid metabolism.We found that treatment with agonistic anti-4-1BB mAb promoted the proliferation of CD8^(+)T cells in vitro,increasing their size and granularity.Studies with a glycolysis inhibitor and a fatty acid oxidation inhibitor revealed that CD8^(+)T cell proliferation required both glucose and fatty acid metabolism.Anti-4-1BB treatment increased glucose transporter 1 expression and activated the liver kinase B1(LKB1)-AMP-activated protein kinase(AMPK)-acetyl-CoA carboxylase(ACC)signaling pathway,which may be responsible for activating the metabolism of glucose and fatty acids.We also examined whether blocking glucose or fatty acid metabolism affected cell cycle progression and the anti-apoptotic effect of 4-1BB signaling.The increase of anti-apoptotic factors and cyclins in response to anti-4-1BB treatment was completely prevented by treating CD8^(+)T cells with the fatty acid oxidation inhibitor,etomoxir,but not with the glycolysis inhibitor,2-deoxy-D-glucose.We conclude that anti-4-1BB treatment activates glucose and fatty acid metabolism thus supporting the increased demand for energy and biomass,and that fatty acid metabolism plays a crucial role in enhancing the cell cycle progression of anti-CD3-activated CD8^(+)T cells in vitro and the anti-apoptotic effects of 4-1BB signaling on these cells.

LIX1-like protein promotes liver cancer progression via miR-21-3p-mediated inhibition of fructose-1,6-bisphosphatase

Limb and CNS expressed 1 like(LIX1L) is over-expressed in several types of tumors.However,the function of LIX1L in glucose metabolism and hepatocellular carcinoma(HCC) progression remains elusive.Here we report that LIX1L is over-expressed in human HCC tissues,which predicts unfavorable prognosis.LIX1L deficiency in vivo significantly attenuated liver cancer initiation in mice.Functional studies indicated that LIX1L overexpression elevated proliferation,migratory,invasive capacities of HCC cells in vitro,and promoted liver cancer growth and metastasis in vivo.LIX1L knockdown up-regulated fructose-1,6-bisphosphatase(FBP1) expression to reduce glucose consumption as well as lactate production.Mechanistically,LIX1L increased miR-21-3p expression,which targeted and suppressed FBP1,thereby promoting HCC growth and metastasis.MiR-21-3p inhibitor could abrogate LIX1L induced enhancement of cell migration,invasion,and glucose metabolism.Inhibition of miR-21-3p suppressed tumor growth in an orthotopic tumor model.Our results establish LIX1L as a critical driver of hepatocarcinogenesis and HCC progression,with implications for prognosis and treatment.

Study on the relationship between relieving energy crisis in myofascial trigger points with An-Pressing manipulation and AMPK/PGC-1α pathway activation

Objective To explore the mechanism of An-Pressing manipulation in relieving energy crisis in chronic myofascial trigger points(MTrPs)by observing the effects of An-Pressing manipulation on adenosine triphosphate(ATP),adenosine 5′-monophosphate(AMP)-activated protein kinase(AMPK)/peroxisome proliferator-activated receptorγcoactivator 1α(PGC-1α)pathway and mitochondrial ultrastructure of skeletal muscle cells in MTrPs rats.Methods Forty-eight male Sprague-Dawley rats were randomly divided into a blank group,a model group,a lidocaine group,and an An-Pressing manipulation group,with 12 rats in each group.The model group,lidocaine group and An-Pressing manipulation group were used to replicate the MTrPs rat model by blunt shock and centrifugal motion method.After modeling,the An-Pressing manipulation group was subjected to 7 times An-Pressing manipulation,once every other day;the lidocaine group was treated with 3 times of injection of lidocaine at the MTrPs,once every 6 d.The blank group and the model group were fed normally without intervention.After the intervention,local muscle tissue was taken to detect the content of ATP and the expression of AMPK,phosphorylated AMPK(phospho-AMPK),PGC-1α,and glucose transporter 4(GluT4),and the ultrastructure of mitochondria was observed under an electron microscope.Results Compared with the blank group,the ATP content in the model group was decreased(P<0.05),the protein expression levels of phospho-AMPK,PGC-1α,and GluT4 and the ratio of phospho-AMPK to AMPK were decreased(P<0.05);under the electron microscope,the number of mitochondria decreased,and they were deformed,small in volume,and had deformed cristae.Compared with the model group,the ATP contents in the An-Pressing manipulation group and the lidocaine group were increased(P<0.05),and the protein expression levels of phospho-AMPK,PGC-1α,and GluT4 and the ratio of phospho-AMPK to AMPK were increased(P<0.05);under the electron microscope,the number of mitochondria increased,the shape and size of the mitochondria were basically normal,and the cristae could be seen.Compared with the lidocaine group,phospho-AMPK and the ratio of phospho-AMPK to AMPK in the An-Pressing manipulation group were increased(P<0.05);under the electron microscope,the numbers of mitochondria were similar,and the shape and size of the mitochondria were basically normal without swelling,and the cristae could be observed.Conclusion An-Pressing manipulation can increase the ATP content in MTrPs tissue,improve the expression levels of PGC-1α and GluT4 proteins and the ratio of phospho-AMPK to AMPK;its mechanism may relate to the activation of AMPK/PGC-1α signaling pathway to promote the repair of mitochondrial damages.

AAZ2 induces mitochondrial-dependent apoptosis by targeting PDK1 in gastric cancer

Drastic surges in intracellular reactive oxygen species(ROS)induce cell apoptosis,while most chemotherapy drugs lead to the accumulation of ROS.Here,we constructed an organic compound,arsenical N-(4-(1,3,2-dithiarsinan-2-yl)phenyl)acrylamide(AAZ2),which could prompt the ROS to trigger mitochondrial-dependent apoptosis in gastric cancer(GC).Mechanistically,by targeting pyruvate dehydrogenase kinase 1(PDK1),AAZ2 caused metabolism alteration and the imbalance of redox homeostasis,followed by the inhibition of phosphoinositide-3-kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)pathway and leading to the activation of B-cell lymphoma 2(Bcl2)/Bcl2-associated X(Bax)/caspase-9(Cas9)/Cas3 cascades.Importantly,our in vivo data demonstrated that AAZ2 could inhibit the growth of GC xenograft.Overall,our data suggested that AAZ2 could contribute to metabolic abnormalities,leading to mitochondrial-dependent apoptosis by targeting PDK1 in GC.

Butein-instigated miR-186-5p-dependent modulation of TWIST1 affects resistance to cisplatin and bioenergetics of Malignant Pleural Mesothelioma cells

Aim:Malignant pleural mesothelioma is a chemoresistant tumor,and biphasic and sarcomatoid histologies portend the worst prognosis for malignant pleural mesothelioma(MPM)patients.We obtained the microRNA expression profile of three biphasic-sarcomatoid MPM cell lines to identify commonly expressed microRNAs and evaluate the effect of butein,a chemo-sensitizing compound,on this microRNA subset.Methods:Nanostring-based microRNA profiling and analysis through the ROSALIND platform were employed to identify the commonly modulated microRNAs and their targets.MicroRNA-mimic transfection,Luciferase assay,and Western blotting were employed to show specific perturbation of TWIST1 levels by miR-186-5p.Sphere-forming assays,invasion assay,and metabolic profiling were used to assess the biological consequences of the butein-instigated miR-186-5p-mediated perturbation of TWIST1 levels.TGCA analysis was used to search for the correlation between TWIST1 and miR-186-5p levels in biphasic and epithelioid MPM specimens.Results:We identified a set of perturbed microRNAs,common to three biphasic/sarcomatoid MPM cell lines,after butein treatment.When focusing on miR-186-5p,we unraveled a butein-ignited and miR-186-5p-mediated modulation of TWIST1 levels which affected the 3D anchorage-independent growth,cisplatin resistance,invasion,and bioenergetics of the MPM cell lines tested.We showed that miR-186-5p and TWIST1 levels are anti-correlated in biphasic MPM specimens from TCGA.Conclusion:We unraveled a novel mechanism of action of butein,which attenuated the pro-tumorigenic features of MPM at least through a miR-186-5p-TWIST1 axis.We suggest that those activities converge into the chemo-sensitizing effect of this compound and may be of translational relevance.

Nicotinamide adenine dinucleotide phosphate promote the rapid growth of the tumor

Nicotinamide adenine dinucleotide phosphate(NADPH) oxidase is the main source of ROS(intracellular reactive oxygen species), ROS plays an important role in a variety of tumor, the ROS mediated by NADPH oxidase increase the expression of hypoxia inducing factor alpha(HIF-α) through multiple signaling pathways in tumor, and HIFcould be regulated and controlled by downstream multiple targeted genes such as vascular endothelial growth factor(VEGF), glucose transporter(GLUT) to promote tumor angiogenesis, cell energy metabolism reprogram and tumor metastasis.HIF-α, meanwhile, also can regulate the expression of NADPH oxidase by ROS, thus further promote development of tumor.In this review, we will summarize the functions of NADPH in tumorigenesis and discuss their potential implications in cancer therapy.

Contribution of Musculoskeletal Disorders to Chronic Lumbago in Parkinson’s Disease

Purpose: To clarify the impact of bone metabolism disorder on lumbago in Parkinson’s Disease (PD). Methods: Data was retrospectively analyzed from 52 patients with PD in our outpatient clinic for more than 1 year (mean age, 63 ± 4 years old;mean duration from onset, 6.3 ± 0.8 years). Patients’ characteristics, comorbid musculoskeletal disorders, serum bone metabolism biomarkers, and bone mineral density were examined. Results: Twenty-one PD patients (40.2%) had chronic lumbago. Severe comptocormia and scoliosis were the most common musculosketal disorders in this group (47.6%) affected by lumbago, followed by osteoporosis (14.3%), compression fracture (4.8%). There was no significant difference in the duration of PD, body mass index, frequency of falls, bone mineral density, tartrate-resistant acid phosphatase-5b, osteocalcin, and N-terminal telopeptide between PD patients with or without chronic lumbago. Multivaritae logistic regression analysis identified the independent predictors of chroni lumbago in PD patients as Hoen-Yahr stage (odds ration [OR] = 2.794, 95%CI 1.103 - 7.076), and elevated serum 1,25-OH2 vitamin D level ([OR] = 0.92, 95%CI 0.86 - 98). Conclusion: Bone metabolism disorders are found to be associated with chronic lumbago in PD patients.

促红细胞生成素改善ob／ob小鼠肝脏脂质沉积

目的研究促红细胞生成素（EPO）对肝脏脂质沉积的影响及可能的分子机制。方法12只雄性8周龄遗传型肥胖小鼠（ob／ob）随机数字表法分为2组：一组腹腔注射EPO（3000UAg），即EPO组（ob/ob＋EPO，n=6）；另一组注射等量磷酸盐缓冲液（PBS），即PBS组（ob／ob＋PBS，n=6）；同窝野生型C57BU6小鼠作为正常对照组（C57BU6，n=6），药物干预5周。以棕榈酸（PA，0．5mmol／L）及不同浓度EPO处理人肝癌细胞株HepG2，分为对照组（不处理）、PA组、PA＋EPOf5U／my组及PA＋EPOf10U／m1）组。油红O染色观察各组小鼠肝组织及HepG2细胞内脂质沉积。Western blotting法检测各组小鼠肝脏及HepG2细胞中EPO受体（EPOR）、固醇调节元件结合蛋白lc（SREBP．1c）、脂肪酸合酶（FAS）、乙酰辅酶A羧化酶（ACC）及肉毒碱棕榈酰基转移酶1a（CPT．1a）的蛋白表达。多组间数据比较采用单因素方差分析，两两比较采用最小显著差异法。结果油红O染色显示EPO干预的小鼠肝脏组织及HepG2细胞内脂质沉积均较各自对照组减少。与PBS组相比，EPO组小鼠肝脏SREBP—1c、FAS、ACC蛋白水平显著下降（分别为0．78~0．08比1．23±0．03、1．03±0．08比1．16±0．01、1．10±0．33比1．69±0．02，t=-6．906、-2．756、-8．794．均P〈0．05），EPOR和CPT．1a蛋白水平明显升高（分别为1．28±0．14比0．97±0．06、0．77~0．06比0．60±0．12，t=2．749、2．655，均P〈0．05）。在HepG2细胞中，与PA组相比，PA＋EPO（10U／m1）组中SREBP．1c、FAS、ACC蛋白水平显著下降（t=-10．731、-2．760、-9．618，均P〈0．05），EPOR和CPT-1a蛋白水平升高（t=7．556、2．674，均P〈0．05）。结论EPO可能通过抑制肝脏脂质合成，促进脂肪酸氧化，从而减少肝脏脂质过度沉积。