Constituents from Terminalia species increase PPARα and PPARγ levels and stimulate glucose uptake without enhancing adipocyte differentiation

Ethnopharmacological relevance

The fruits of Terminalia bellerica Roxb. (Combretaceae) and T. chebula Retz. (Combretaceae) are important components of triphala, a popular Ayurvedic formulation, for treating diabetes in Indian traditional medicine.

Aim of the study

The aim of this study was to evaluate the effects of the constituents of T. bellerica and T. chebula fruit extracts on PPARα and PPARγ signaling/expression, cellular glucose uptake and adipogenesis.

Materials and methods

PPARα and PPARγ signaling and expression (luciferase assay and western blot) and the insulin-stimulated uptake of 2-NBDG were determined in HepG2 cells. The effects on adipogenesis were determined in 3T3-L1 cells by Oil red O staining and measurement of lipid content by AdipoRed reagent.

Results

Out of the 20 compounds, two ellagitannins, chebulagic acid (1) and corilagin (2), and three gallotannins, 2,3,6-tri-O-galloyl-β-d-glucose (3), 1,2,3,6-tetra-O-galloyl-β-d-glucose (4), and 1,2,3,4,6-penta-O-galloyl-β-d-glucose (5), showed the enhancement of PPARα and/or PPARγ signaling. Two of the gallotannins (4 and 5) also increased PPARα and PPARγ protein expression, while all three (3–5) enhanced insulin-stimulated glucose uptake into HepG2 cells. Compound 1,2,3,6-tetra-O-galloyl-β-d-glucose (4) was the most potent in increasing cellular glucose uptake (9.92-fold increase at 50 μM). In the test for adipogenesis, 3–5 did not enhance the differentiation of 3T3-L1 preadipocytes but inhibited the adipogenic effect of rosiglitazone.

Conclusion

Three gallotannins (3–5) from Terminalia fruits acting as enhancers of both PPARα and PPARγ signaling increased insulin-stimulated glucose uptake without inducing the adipogenesis, with 1,2,3,6-tetra-O-galloyl-β-d-glucose (4) being the most effective in stimulating glucose uptake and 1,2,3,4,6-penta-O-galloyl-β-d-glucose (5) being most effective in increasing PPAR protein expression.     

Methanolic extract of Momordica cymbalaria enhances glucose uptake in L6 myotubes in vitro by up-regulating PPAR-γ and GLUT-4

The present study was undertaken to evaluate the influence of the methanolic fruit extract of Momordica cymbalaria （MFMC） on PPARγ, （Peroxisome Proliferator Activated Receptor gamma） and GLUT-4 （Glucose transporter-4） with respect to glucose transport. Various concentrations of MFMC ranging from 62.5 to 500 μg·mL^-1 were evaluated for glucose uptake activity in vitro using L6 myotubes, rosiglitazone was used as a reference standard. The MFMC showed significant and dose-dependent increase in glucose uptake at the tested concentrations, further, the glucose uptake activity of MFMC （500 μg·mL^-1） was comparable with rosigilitazone. Furthermore, MFMC has shown up-regulation of GLUT-4 and PPARy gene expressions in L6 myotubes. In addition, the MFMC when incubated along with cycloheximide （CHX）, which is a protein synthesis inhibitor, has shown complete blockade of glucose uptake. This indicates that new protein synthesis is required for increased GLUT-4 translocation. In conclusion, these findings suggest that MFMC is enhancing the glucose uptake significantly and dose dependently through the enhanced expression of PPARμ and GLUT-4 in vitro.     

Activation of the nuclear receptor PPARγ by metabolites isolated from sage (Salvia officinalis L.)

Ethnopharmacological relevance

Salvia officinalis has been used as a traditional remedy against diabetes in many countries and its glucose-lowering effects have been demonstrated in animal studies. The active compounds and their possible mode of action are still unknown although it has been suggested that diterpenes may be responsible for the anti-diabetic effect of Salvia officinalis.

Aim of the study

To investigate whether the reported anti-diabetic effects of Salvia officinalis are related to activation of the nuclear receptor peroxisome proliferator-activated receptor (PPAR)γ and to identify the bioactive constituents.

Materials and methods

From a dichloromethane extract of Salvia officinalis able to activate PPARγ several major metabolites were isolated by chromatographic techniques. To assess bioactivity of the isolated metabolites a PPARγ transactivation assay was used.

Results

Eight diterpenes were isolated and identified including a new abietane diterpene being the epirosmanol ester of 12-O-methyl carnosic acid and 20-hydroxyferruginol, which was isolated from Salvia officinalis for the first time, as well as viridiflorol, oleanolic acid, and α-linolenic acid. 12-O-methyl carnosic acid and α-linolenic acid were able to significantly activate PPARγ whereas the remaining metabolites were either unable to activate PPARγ or yielded insignificant activation.

Conclusions

Selected metabolites from Salvia officinalis were able to activate PPARγ and hence, the anti-diabetic activity of this plant could in part be mediated through this nuclear receptor.     

Isoimperatorin, cimiside E and 23-O-acetylshengmanol-3-xyloside from Cimicifugae rhizome inhibit TNF-α-induced VCAM-1 expression in human endothelial cells: involvement of PPAR-γ upregulation and PI3K, ERK1/2, and PKC signal pathways

Ethnopharmacological relevance

The methanol extract of Cimicifugae Rhizome has been traditionally used in various disorders including inflammation.

Aim of the study

The aim of the study is to explore whether anti-inflammatory action of 3 active compounds, two triterpenoid glycosides (cimiside E, 23-O-actylshengmanol-3-xyloside) and one furanocoumarin (isoimperatorin), isolated from Cimicifugae Rhizome is related with peroxisome proliferator-activated receptor-γ (PPAR-γ) expression in human umbilical endothelial cell line, EA.hy926 cells.

Materials and methods

Cell viability and production of reactive oxygen species were performed. In addition, adhesion of monocyte into endothelial cells and western blot for expression of adhesion molecules and signal proteins were investigated in tumor necrosis factor-α (TNF-α)-activated cells.

Results

Pretreatment of test compounds significantly reduced reactive oxygen species (ROS) production and expression of vascular cell adhesion molecule-1 (VCAM-1), but not intercellular cell adhesion molecule-1 (ICAM-1). Three compounds all dose-dependently increased not only PPAR-γ expression in EA.hy926 cells but inhibited TNF-α-induced phosphorylation of Akt, extracelullar-signal-regulated kinase (ERK) and protein kinase C (PKC) with different specificity. Finally, they prevented TNF-α-induced adhesion of U937 monocytic cells to EA.hy926 cells.

Conclusions

The present results show that cimiside E, 23-O-actylshengmanol-3-xyloside, isoimperatorin isolated from Cimicifugae Rhizome selectively inhibits TNF-α-induced expression of VCAM-1 at least by upregulation of PPAR-γ, and signals for ERK1/2, PI3K, and PKC are involved in this effect.     

Wild bitter gourd extract up-regulates mRNA expression of PPARα, PPARγ and their target genes in C57BL/6J mice

Ethnopharmacological relevance

Wild bitter gourd (Momordica charantia Linn. var. abbreviata ser.) was commonly used as a medicinal herb in Asia, Africa, and South America because of its anti-diabetic, antibacterial, anti-viral, and chemopreventive functions.

Materials and methods

C57BL/6J mice were orally administered with 250, 500 or 1000 mg/kg BW of WBGE in 0.2 mL/mouse of olive oil daily for 2 weeks.

Results

Compared to control (vehicle treated) mice, mice receiving WBGE showed significantly higher PPARα, ACO (acyl-CoA oxidase) and L-FABP (liver-fatty acid binding protein) mRNA expression, ACO activity and protein in the liver (P < 0.05), as clofibrate-treated mice. WBGE treatment also resulted in significantly higher PPARγ and LPL (lipoprotein lipase) mRNA (P < 0.05) in the epididymal adipose tissue. Liver triglyceride and non-esterified fatty acid concentration in WBGE treated mice were significantly lower than those of control mice (P < 0.05). Plasma adiponectin level was significantly higher in mice receiving WBGE than in control mice (P < 0.05), as the rosiglitazone treated mice.

Conclusion

Results of this study demonstrated that WBGE also activates PPARα and PPARγ signaling pathway in vivo.     

A UPLC–MS/MS method for in vivo and in vitro pharmacokinetic studies of psoralenoside,isopsoralenoside, psoralen and isopsoralen from Psoralea corylifolia extract

Ethnopharmacological relevance

The dried fruit of Psoralea corylifolia L. has been used to prevent and treat vitiligo, osteoporosis, arthralgia and asthma in Traditional Chinese Medicine for some 1600 years. Psoralen (P), isopsoralen (IP), psoralenoside (PO) and isopsoralenoside (IPO) are the major coumarins and coumarin-related benzofuran glycosides in Psoraleae Fructus, which have been reported to show estrogen-like activity, osteoblastic proliferation accelerating activity, antitumor effects and antibacterial activity. The first aim of this study is to develop a rapid, sensitive and selective ultra performance liquid chromatography tandem mass spectrometry (UPLC–MS/MS) approach for simultaneous determination of PO, IPO, P and IP in rat plasma and samples collected from in vitro incubation experiments. The second aim is to investigate the pharmacokinetic properties of PO, IPO, P and IP after oral administration of Psoralea corylifolia extract (PCE) to rats. The third aim is to confirm the biotransformation of PO to P or IPO to IP under gastrointestinal conditions.

Materials and methods

A UPLC–MS/MS method with a C18 column and a mobile phase of methanol-0.1% aqueous formic acid was validated according to the criteria in FDA guidelines about bioanalytical method, which was developed to investigate the pharmacokinetic behavior of PO, IPO, P and IP from PCE and the metabolic pathways of PO to P or IPO to IP.

Results

The criteria for establishment of a new UPLC–MS/MS method including selectivity, linearity, accuracy, precision, extraction recovery, matrix effect and stability were validated. This method was successfully applied to the quantitative determination of PO, IPO, P and IP in biological samples collected from both in vitro incubations and in vivo rat experiments. After oral administration of PCE to rat, pharmacokinetic parameters of these four compounds indicated that in vivo biotransformation may occur between PO and P or IPO and IP. Purified benzofuran glycosides fraction (PBGF), containing only PO and IPO, was orally administered to rats to further confirm the biotransformation of PO to P or IPO to IP under gastrointestinal conditions. An in vitro incubation study elucidated that PO and IPO were metabolized to P and IP by intestinal microflora through de-glucosylation.

Conclusions

This paper developed a rapid, sensitive and selective UPLC–MS/MS method for simultaneous determination of PO, IPO, P and IP from PCE in biological samples, and investigated on their comprehensive in vivo and in vitro pharmacokinetic studies. These obtained results showed that the metabolism by intestinal bacteria plays an important role in pharmacological effects of orally administered PCE.     

Inhibition of TNF-α-Induced Inflammation by andrographolide via down-regulation of the PI3K/Akt signaling pathway

Andrographolide (1), an active constituent of Andrographis paniculata, decreased tumor necrosis factor-α (TNF-α)-induced intercellular adhesion molecule-1 (ICAM-1) expression and adhesion of HL-60 cells onto human umbilical vein endothelial cells (HUVEC), which are associated with inflammatory diseases. Moreover, 1 abolished TNF-α-induced Akt phosphorylation. Transfection of an activated Akt1 cDNA vector increased Akt phosphorylation and ICAM-1 expression like TNF-α. In addition, 1 and LY294002 blocked TNF-α-induced IκB-α degradation and nuclear p65 protein accumulation, as well as the DNA-binding activity of NF-κB. Compound 1 exhibits anti-inflammatory properties through the inhibition of TNF-α-induced ICAM-1 expression. The anti-inflammatory activity of 1 may be associated with the inhibition of the PI3K/Akt pathway and downstream target NF-κB activation in HUVEC cells.     

The effect of the extract of Crocus sativus on tracheal responsiveness and plasma levels of IL-4, IFN-γ, total NO and nitrite in ovalbumin sensitized Guinea-pigs

Ethnomedical relevance

Anti-inflammatory, anti oxidant and effect of Crocus sativus (C. sativus) on Th₁/Th₂ balance were described previously.

Aim of the study

The preventive effects of the extract of Crocus sativus on tracheal responsiveness and plasma levels of IL-4, IFN-γ, total NO and nitrite were examined on sensitized guinea pigs.

Materials and methods

Five groups of sensitized guinea pigs to ovalbumin (OVA), were given drinking water containing three concentrations of the extract of Crocus sativus, dexamethasone (S+D) or alone (group S). Tracheal responses (TR) of control animals (group C) and sensitized guinea pigs (n=6, for each group) to methacholine, OVA and the levels of IL-4, IFN-γ, total NO and nitrite in serum were examined.

Results

The TR to both methacholine and OVA, the levels of serum IL-4, total NO and nitrite in S guinea pigs were significantly increased but that of IFN-γ and IFN-γ/IL-4 ratio (Th₁/Th₂ balance) were decreased compared to the controls (p<0.05 to p<0.001). In the treated animals with dexamethasone and all concentrations of the extract, TR to both methacholine and OVA, IL-4, total NO and nitrite were significantly decreased but IFN-γ and IFN-γ/IL-4 ratio increased compared to S group (p<0.05 to p<0.001). The effects of the highest concentration of the extract was greater than those of other concentrations and the effect of dexamethasone (p<0.05 to p<0.01).

Conclusions

These results not only showed a preventive effect of C. sativus extract on tracheal responses and serum levels of inflammatory mediators in sensitized guinea pigs but also showed increased Th₁/Th₂ balance.     

Pharmacokinetics,brain distribution,release and blood–brain barrier transport of Shunaoxin pills

Ethnopharmacological relevance

Shunaoxin pills, a traditional Chinese medicine (TCM) product, have been used to treat cerebrovascular diseases in China since 2005. The main active components of Shunaoxin pills are ferulic acid and ligustilide from Chuanxiong (Ligusticum chuanxiong Hort, Umbelliferae) and Danggui (Angelica sinensis radix, Umbelliferae). As Shunaoxin shows excellent activity in the central nervous system (CNS), the extent to which the major constituents of Shunaoxin reach the CNS should be investigated. Moreover, the in vivo–in vitro correlations (IVIVC) of the formulation should be studied to elucidate the mechanisms of action of TCM in the CNS. However, these data have not previously been available. Thus we intended to investigate what the extent when these constituents of Shunaoxin pills reach the CNS, and evaluate the IVIVC of release and pharmacokinetics.

Materials and methods

In this study, we evaluated the release of ferulic acid and ligustilide from Shunaoxin pills, and their transport across an in vitro model of the BBB. We also evaluated their pharmacokinetics and brain distribution in vivo. High-performance liquid chromatography (HPLC) was used to quantify both compounds simultaneously. Based on the release in vitro and absorption of ferulic acid and ligustilide in vivo, IVIVC permitted prediction of the pharmacokinetics of these compounds.

Results

The release of ferulic acid and ligustilide reached a platform phase within 1 h. Ferulic acid and ligustilide rapidly crossed the BBB in different patterns; the transport ratio increased over time. After intragastric (i.g.) administration of Shunaoxin pills, ferulic acid and ligustilide were rapidly absorbed and distributed into brain, which may result in a rapid onset of action.

Conclusions

Ferulic acid and ligustilide were transported across a model BBB. After i.g. administration of Shunaoxin pills, ferulic acid and ligustilide were rapidly absorbed and distributed in brain; this may lead to rapid pharmacological onset. The IVIVC can be used to predict in vivo pharmacokinetics from in vitro experimental results. These results provide support for the clinical use of Shunaoxin pills.     

桑叶提取物调控IRS-1/PI3K/GLUT4通路影响2型糖尿病胰岛素抵抗机制研究

目的:提取桑叶中的黄酮类及多酚类物质,观察其对2型糖尿病(T2DM)模型大鼠降血糖及改善胰岛素抵抗(IR的作用,并探讨其分子机制。方法:取雄性SD大鼠,分为正常组、模型组、二甲双胍组、桑叶提取物组,除正常组以外建立T2DM模型,灌胃干预4周,观察药物对模型大鼠一般情况、生化指标、骨骼肌病理等的影响。测定胰岛素受体底物-1 (IRS-1)、磷脂酰肌醇3-激酶(PI3K)中的p85α、葡萄糖转运体-4 (GLUT4)在骨骼肌组织中的信使核糖核酸(mRNA)及蛋白表达水平。结果:与正常组比较,模型组空腹血糖(FBG)、IR稳态模型指数(HOMA-IR)、血清胰岛素、总胆固醇、甘油三酯、低密度脂蛋白水平升高(P<0.05);IRS-1、PI3K p85α、GLUT4的mRNA及蛋白表达明显下降(P<0.05)。与模型组比较,桑叶提取物组FBG、HOMA-IR、总胆固醇、甘油三酯、低密度脂蛋白水平下降(P<0.05);IRS-1、PI3K p85α、GLUT4的mRNA及蛋白表达明显升高(P<0.05);二甲双胍组FBG有所下降,但差异无统计学意义,血清胰岛素及HOMA-...     

昆仑雪菊提取物对糖尿病大鼠胰岛素抵抗IRS-1/PI3K/GLUT4信号通路的影响

目的:观察昆仑雪菊提取物对链脲佐菌素(STZ)诱导的糖尿病大鼠血糖、胰岛素分泌水平及胰岛素受体底物-1/磷脂酰肌醇(-3)激酶/葡萄糖转运蛋白4(IRS-1/PI3K/GLUT4)信号传导通路的影响。方法:将35只雄性STZ诱导的糖尿病Wistar大鼠随机分为5组,模型组、昆仑雪菊提取物低、中、高剂量组、吡咯列酮组,每组7只;另选7只SPF级雄性Wistar大鼠为正常组。正常组和模型组灌服生理盐水,昆仑雪菊低、中、高剂量组,分别按100,200,400 mg·kg~(-1)·d~(-1)剂量灌胃,吡咯列酮组灌服吡咯列酮10 mg·kg~(-1)·d~(-1)。连续4周。4周后,心包外抽血,离心取血清;取胰腺组织。采用酶联免疫吸附(ELISA)法检测大鼠胰岛素(INS)分泌水平;蛋白免疫印迹(Western blot)法检测骨骼肌IRS-1,PI3K,GLUT4的蛋白表达;实时荧光定量聚合酶链式反应(Real-time PCR)检测骨骼肌GLUT4的mRNA表达。结果:灌胃4周后,经检测发现昆仑雪菊提取物对降低糖尿病大鼠体重作用不明显,但对空腹血糖、餐后2 h血糖具有降低作用,并在一定程度上能够促进胰岛素分泌,促进调节因子IRS-1,PI3K,GLUT4的表达,与模型组比较有显著性差异(P0.05)。结论:昆仑雪菊提取物可降低2型糖尿病大鼠血糖,促进胰岛素分泌,提高大鼠骨骼肌IRS-1,PI3K,GLUT4的表达,改善胰岛素抵抗。     

Magnolol and 5-fluorouracil synergy inhibition of metastasis of cervical cancer cells by targeting PI3K/AKT/mTOR and EMT pathwaysOA

Objective: This study is designed to investigate the mode of action of the synergistic effect of 5-fluorouracil(5-FU) and magnolol against cervical cancer.Methods: Network pharmacological approach was applied to predict the molecular mechanism of 5-FU combined with magnolol against cervical cancer. CCK-8 assay, colony formation assay, immunofluorescence staining, adhesion assay, wound healing mobility assay, cell migration and invasion assay and Western blot analysis were conducted to validate t...     

The impact of Zataria multiflora Boiss extract on in vitro and in vivo Th1/Th2 cytokine (IFN-γ/IL4) balance

Ethnopharmacological relevance

Anti-inflammatory, anti-oxidant and effect of Zataria multiflora on Th₁/Th₂ balance were previously described. Different therapeutic effects of this plant have been described in Iranian traditional medicine. To evaluate the immune modulatory effects of Zataria multiflora on Th1/Th2 balance, which may be implicated in inflammatory disorders, in vitro and in vivo studies were carried out.

Materials and methods

The effects of three concentrations of the extract, dexamethasone, and saline on interleukin 4 (IL-4) and interferon γ (IFN-γ) gene expression were evaluated in phytohemagglutinin (PHA) stimulated and non-stimulated human peripheral blood mononuclear cells (hPBMCs). RNA was extracted from the hPBMCs to make cDNA for real time PCR relative quantification. Furthermore, the effect of the extract on serum level of IL-4 and IFN-γ was assessed in ovalbumin (OA) sensitized guinea pigs (n=6 for each group).

Results

Dexamethasone showed significant inhibitory effect on both IFN-γ and IL-4 gene expression and serum level of the cytokines and significantly enhanced IFN-γ/IL-4 ratio (p<0.05–p<0.001). The extract inhibited IL-4 and enhance IFN-γ gene expression and IFN-γ/IL-4 ratio too (p<0.05–p<0.001). In sensitized animals also serum level of IL-4 were significantly decreased after treatment with both dexamethasone and extract, but serum level of IFN-γ and IFN-γ/IL-4 ratio were significantly increased due to extract treatment (p<0.01 for medium and p<0.001 for high concentration).

Conclusions

These results indicated consistent in vitro and in vivo data for selective immune modulatory effect of the extract of Zataria multiflora which increased IFN-γ, decreased IL-4, and enhanced the ratio of IFN-γ to IL-4 (Th₁/Th₂ balance). Therefore, the extract of Zataria multiflora may have therapeutic value in inflammatory responses such as allergy, autoimmunity and infectious diseases associated with Th₁/Th₂ imbalance.     

基于TLR4/NF-κB/NLRP3和PI3K/Akt通路研究甘遂及其炮制品对水负荷小鼠的利尿作用及机制

目的 研究甘遂及其炮制品对水负荷小鼠的利尿作用及机制。方法 通过网络药理学探究甘遂的利尿作用机制。ICR小鼠给予甘遂及其炮制品后,ig大量生理盐水建立水负荷模型,测定水负荷差值及脏器指数;取十二指肠及空肠,观察病理变化;采用ELISA法测定小鼠血清中炎性因子指标白细胞介素-1β(interleukin-1β,IL-1β)、IL-18、IL-4、IL-10及小肠组织中氧化应激指标丙二醛（malondialdehyde,MDA）和还原型谷胱甘肽（glutathione,GSH）水平;采用Western blotting测定小鼠小肠组织中Toll样受体4(Toll-like receptor 4,TLR4)、核因子-κB(nuclear factor-κB,NF-κB)、NOD样受体蛋白3(NOD-like receptor protein 3,NLRP3)、磷脂肌醇-3-激酶（phosphatidylinositol-3-kinase,PI3K）和蛋白激酶B(protein kinase B,Akt)蛋白表达。结果 通过构建“中药-成分-疾病-通路-靶点”网络及通路富集分析得到甘遂及其炮制...     

A combination extract of Renshen (Panax Ginseng), Yinyanghuo (Herba Epimedii Brevicornus), Yuanzhi (Radix Palygalae) and Jianghuang (Rhizoma Curcumae Longae) decreases glycogen synthase kinase 3β expression in brain cortex of APPV717I transgenic mice

Objective

To investigate the neuroprotective mechanism of combination extract of Renshen (Panax Ginseng), Yinyanghuo (Herba Epimedii Brevicornus), Yuanzhi (Radix Palygalae) and Jianghuang (Rhizoma Curcumae Longae) (GEPT) in treating Alzheimer's disease on the target of glycogen synthase kinase 3β (GSK-3β).

Methods

Three-month-old APPV717I transgenic mice were randomly divided into ten groups (n=12 per group) and intragastrically administrated vehicle or medicines: APP group was given 0.5% CMC, donepezil group was given donepezil (APP + D group) (0.92 mg/kg⁻¹ · day⁻¹), and GEPT groups were given small dose of GEPT (APP+Gs group) (0.075 g/kg⁻¹ · day⁻¹), medium dose (APP+Gm group) (0.15 g/kg⁻¹ ·day⁻¹), and large dose (APP+Gl group) (0.30 g/kg⁻¹ · day⁻¹) for 4 or 8 months, respectively. Three-month-old C57BL/6J mice as vehicle controls (n=12) were given 0.5% CMC for 4 or 8 months as well. The GSK-3β expression in the cortex of 7- and 11-month-old APPV717I transgenic mice with and without GEPT or donepezil treatment and normal C57BL/6J mice were measured via Western blotting and Immunohistochemistry.

Results

Immunohistochemistry analysis showed significant increase of GSK-3β in the cerebral cortex of 7-month-old APP group (compare to control group P=0.003), while the GSK-3β expression of donepezil or GEPT group were all significantly decreased (Donepezil vs APP: P=0.041; Gl vs APP: P= 0.049; Gm vs APP: P=0.029; Gh vs APP: P=0.036). Western blot analysis showed similar results. The densitometric measures of GSK-3β in APP mice increased significantly as compared with the control group (P=0.008). And the GSK-3β expression in donepezil and GEPT groups were all decreased. There was significant difference between Gh group or donepezil group and the control group (P=0.05). Similar findings were shown in the 11-month-old mice in each group, except for greater decrease of GSK-3β in the GEPT group.

Conclusion

GEPT can effectively decrease the level of GSK-3β expression in the brain cortex of AP-PV717I transgenic mice, and such effect is more significant in 11-month-old mice. This partially explains the neuroprotecting mechanism of GEPT in preventing and treating of AD.     

坎离颗粒对血管紧张素II阻断骨骼肌细胞糖代谢PI3K/PKB/GLUT-4信号途径的影响及机制探讨

目的:探究坎离颗粒对小鼠骨骼肌成肌细胞(C2C12细胞)糖代谢的影响;探明坎离颗粒对血管紧张素Ⅱ阻断C2C12细胞糖代谢的影响是否通过PI3K/PKB/GLUT-4信号途径。方法:以C2C12细胞为研究对象,以胰岛素激活细胞糖代谢通路,以血管紧张素Ⅱ阻断被激活的通路,在此基础上设胰岛素组、模型对照组、坎离颗粒2、20、200μg/ml组,另设空白对照组。药物干预结束后,检测细胞中乳酸脱氢酶(LDH)及同工酶、乳酸、三磷酸腺苷(ATP)含量,葡萄糖运载体4(GLUT-4)、蛋白激酶B(PKB)、磷脂酰肌醇3激酶(PI3K)、丝裂原活化蛋白激酶p38(P38MAPK)蛋白及mRNA表达。结果:与空白组比较,胰岛素组对荧光标记2-脱氧葡萄糖(2-NBDG)摄取率显著升高;与胰岛素组比较,模型组对2-NBDG摄取率显著降低;与模型组比较,坎离颗粒三个剂量组对2-NBDG摄取率均显著升高。坎离颗粒2μg/ml组可明显降低乳酸水平。坎离颗粒20、200μg/ml组均可明显降低LDH4水平。与胰岛素组比较,模型组GLUT-4蛋白及mRNA、P-PKB、P-PI3K蛋白表达显著下调;与模型组比较,坎离颗粒可上调GLUT-4蛋白及mRNA、P-PKB、P-PI3K蛋白表达,以20、200μg/ml组最显著。结论:坎离颗粒可促进C2C12细胞糖摄取,改善糖代谢。并通过上调GLUT-4蛋白和mRNA表达、PKB、PI3K磷酸化蛋白表达,改善C2C12细胞的糖代谢,坎离颗粒对血管紧张素Ⅱ阻断骨骼肌细胞糖代谢的改善作用是通过PI3K/PKB/GLUT-4信号通路。     

大黄蛰虫丸调控ROS介导的PI3K/Akt/FoxO4信号通路改善大鼠肝脏衰老的作用和机制北大核心CSCD

近年来的研究显示,非酒精性脂肪性肝病、肝硬化以及肝癌等常见肝病的发生和发展都与肝脏衰老(liver aging,LA)有关。基于此,为了探究传统经方——大黄蛰虫丸(Dahuang Zhechong Pills,DHZCP)多靶点地改善LA的作用和机制,笔者将24只大鼠随机均分为4组,即正常组、衰老模型组(模型组)、衰老模型+DHZCP组(DHZCP组)和衰老模型+维生素E(vitamin E,VE)组(VE组)。通过D-半乳糖(D-galactose,D-gal)连续腹腔注射建立大鼠LA模型。对于LA模型鼠,采用衰老表型和体质量来评估其一般情况;采用肝细胞衰老病理学特征、肝功能指标、肝脏磷酸化组蛋白H2A变异体(phosphorylated histone family 2A variant,γ-H2AX)染色特征、肝脏细胞周期阻滞蛋白(P21、P53、P16)表达水平以及肝脏衰老相关分泌表型(senescence-associated secretory phenotypes,SASP)因子蛋白表达水平来评估其LA;采用肝脏活性氧产物(reactive oxygen species,ROS)表达特征以及肝组织磷脂酰肌醇-3激酶(phosphatidylinositol-3 kinase,PI3K)/蛋白激酶B(protein kinase B,Akt)/叉头盒蛋白O4(forkhead box protein O4,FoxO4)信号通路关键信号分子蛋白表达水平来评估ROS介导的PI3K/Akt/FoxO4信号通路活性。结果显示,经DHZCP或VE干预12周后,对于DHZCP组和VE组大鼠,其特征性衰老表型和体质量,肝细胞衰老病理学特征和肝功能指标,肝脏ROS相对表达量,肝组织磷酸化PI3K(phosphorylated PI3K,p-PI3K)、磷酸化Akt(phosphorylated Akt,p-Akt)、FoxO4等关键信号分子的蛋白表达水平,肝脏γ-H2AX染色的特征以及肝组织P16、P21、P53和白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的蛋白表达水平均得到相应改善,且两者的作用相似。总之,借助D-gal诱导的LA模型鼠,该研究阐明,DHZCP在体内能多靶点地改善LA,其作用和机制与调控肝组织ROS介导的PI3K/Akt/FoxO4信号通路活性有关。这些发现为DHZCP治疗衰老相关肝病提供了新的药理学证据。