OBJECTIVETo explore the mechanism of the protective effects of Panax notoginseng saponins (PNS) on kidney in diabetic rats.

METHODSDiabetic rat model was obtained by intravenous injection of alloxan, and the rats were divided into model, PNS-100 mg/(kg day) and PNS-200 mg/(kg day) groups, 10 each. Another 10 rats injected with saline were served as control. Periodic acid-Schiff staining and immunological histological chemistry were used to observe histomorphology and tissue expression of bone morphogenetic protein-7 (BMP-7). Silent information regulator 1 (SIRT1) was silenced in rat mesangial cells by RNA interference. The mRNA expressions of SIRT-1, monocyte chemoattractant protein-1 (MCP-1), transforming growth factor β1 (TGF-β1) and plasminogen activator inhibitor-1 (PAI-1) were analyzed by reverse transcription polymerase chain reaction. The protein expressions of SIRT1 and the acetylation of nuclear factor κB (NF-κB) P65 were determined by western blotting. The concentration of MCP-1, TGF-β1 and malondialdehyde (MDA) in culture supernatant were detected by enzyme-linked immuno sorbent assay. The activity of superoxide dismutase (SOD) was detected by the classical method of nitrogen and blue four.

RESULTSIn diabetic model rats, PNS could not only reduce blood glucose and lipid (P<0.01), but also increase protein level of BMP-7 and inhibit PAI-1 expression for suppressing fibrosis of the kidney. In rat mesangial cells, PNS could up-regulate the expression of SIRT1 (P<0.01) and in turn suppress the transcription of TGF-β1 (P<0.05) and MCP-1 (P<0.05). PNS could also reverse the increased acetylation of NF-κB p65 by high glucose. In addition, redox regulation factor MDA was down-regulated (P<0.05) and SOD was up-regulated (P<0.01), which were both induced by SIRT1 up-regulation.

CONCLUSIONSPNS could protect kidney from diabetes with the possible mechanism of up-regulating SIRT1, therefore inhibiting inflammation through decreasing the induction of inflammatory cytokines and TGF-β1, as well as activating antioxidant proteins.

Acetylation ; drug effects ; Animals ; Antioxidants ; metabolism ; Blood Glucose ; metabolism ; Bone Morphogenetic Protein 7 ; metabolism ; Chemokine CCL2 ; metabolism ; Diabetes Mellitus, Experimental ; blood ; drug therapy ; genetics ; physiopathology ; Gene Knockdown Techniques ; Immunohistochemistry ; Kidney ; drug effects ; pathology ; Kidney Function Tests ; Lipids ; blood ; Male ; Malondialdehyde ; metabolism ; Mesangial Cells ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Panax notoginseng ; chemistry ; Plasminogen Activator Inhibitor 1 ; genetics ; metabolism ; Protective Agents ; pharmacology ; therapeutic use ; Rats, Sprague-Dawley ; Saponins ; pharmacology ; therapeutic use ; Sirtuin 1 ; genetics ; Superoxide Dismutase ; metabolism ; Transcription Factor RelA ; metabolism ; Transcription, Genetic ; drug effects ; Transforming Growth Factor beta1 ; metabolism ; Up-Regulation ; drug effects

PURPOSE: Rheumatoid arthritis (RA) is an inflammatory joint disorder, the progression of which leads to the destruction of cartilage and bone. Chemokines are involved in RA pathogenesis. In this study, we investigated the chemokine signaling pathway associated with CCL2 in peripheral blood (PB) and synovial tissues (ST) of RA patients based on our previous work about chemokine signaling pathway involved in the activation of CCL2 production in collagen-induced arthritis rat ST. MATERIALS AND METHODS: Total RNA was isolated from PB leukocytes and synovium of the knee joint in both RA patients and control populations. Real-time polymerase chain reaction was used to determine CCL4, CCR5, c-Jun, c-Fos, and CCL2 expressions. Serum level of CCL2 was assessed by enzyme-linked immunosorbent assay, and the production of CCL2 in ST was analyzed immunohistochemically. RESULTS: The expressions of CCL4, CCR5, c-Jun, c-Fos, and CCL2 messenger RNA in RA patients were significantly higher than those in healthy controls, both in ST and on PB leukocyte. Serum CCL2 levels were elevated in RA patients. Histological examination of rheumatoid joints revealed extensive CCL2 expression in RA ST. CONCLUSION: CCL2, CCL4, c-Jun, c-Fos, and CCR5 may play an important role in the recruitment of PB leukocytes into the RA joints. These data provide evidence that the chemokine signaling pathway is involved in CCL2 expression in RA patient tissues, which may contribute to chronic inflammation associated with RA. Targeting this signaling pathway may provide a novel therapeutic avenue in RA.
Adult ; Animals ; Arthritis, Rheumatoid/*blood/metabolism ; Case-Control Studies ; Chemokine CCL2/*blood/metabolism ; Chemokines/metabolism ; Enzyme-Linked Immunosorbent Assay ; Female ; Gene Expression ; Humans ; Male ; Middle Aged ; RNA, Messenger/genetics/metabolism ; Rats ; Real-Time Polymerase Chain Reaction ; Signal Transduction ; Synovial Membrane/*metabolism

OBJECTIVETo evaluate the protective effects of punicosides on alcohol induced acute liver injury in mice and its possible mechanisms as well.

METHODThe 60 mice were randomly divided into normal control, model group, three dose groups of punicosides with low, medium and high, then there is silibinin group. Three dose groups of punicosides and silibinin were given in advance by gavage for 4 weeks, then the mouse model of alcoholic acute liver injury was established. The serum levels of ALT, AST and TG were determined, and the mice were killed to calculate somatic index of liver, thymus as well as spleen. MDA, SOD, GSH-Px and GSH-ST were detected in the liver homogenate. Histopathological changes of the liver were observed by HE staining. The expression of MCP-1 and NF-kappaB in the liver tissues were detected by immunohistochemistry.

RESULTMid and high dose of punicosides reduced the liver index in mice significantly, improved liver steatosis, decreased the level of ALT, AST and TG in serum and the content of MDA in liver homogenate, furthermore the two dose groups increased the activity of SOD, GSH-Px and GSH-ST, inhibited the expression of MCP-1 and NF-kappaB in liver tissue.

CONCLUSIONPunicosides can protect the acute liver damage induced by alcohol.

Alcohols ; adverse effects ; Animals ; Chemical and Drug Induced Liver Injury ; blood ; drug therapy ; metabolism ; pathology ; Chemokine CCL2 ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Gene Expression Regulation ; drug effects ; Liver ; drug effects ; enzymology ; pathology ; Male ; Malondialdehyde ; metabolism ; Mice ; NF-kappa B ; metabolism

This study was purposed to investigate the accumulative regularity of tumor-associated noncellular components in supernatant of stored packed red blood cells (PRBC) during storage. The supernatant of PRBC was obtained by centrifugation with 1 006×g for 10 min at day 0, 7, 14, 21, 28 and 35 d. The enzyme-linked immunosorbent assay (ELISA) was used to detect the expression of T cells and the accumulative levels of secreted RANTES/CCL5, tumor necrosis factor-alpha (TNF-α), platelet derived growth factor (PDGF), vascular endothelial growth factor (VEGF) and monocyte chemotactic protein-1 (MCP-1). The results showed that the high concentration of RANTES/CCL5 and TNF-α was found in fresh PRBC, and their accumulative concentration did not increase along with the prolonging of storage time. The VEGF level in PRBC at day 0 of storage was 229.9 pg/ml, and increased to 749.08 pg/ml at end of day 7, then it was stable, and increased to 760.67 pg/ml at end of day 35. The PDGF level in supernatant of PRBC was 13.54 ng/L at dag 0 of storage, and increased stably during storage, then decreased at day 28, however PDGF rapidly rose to 22.13 ng/L at the end of day 35 (P < 0.05). The MCP-1 level in supernatant of PRBC was 39.98 ng/L at day 0 of storage, then slowly increased during storage time, at end of day 35 MCP-1 level increased to 49.83 ng/L. It is concluded that along with the prolongation of storage time, the growth factors in the supernatant of PRBC display the tendency of accumulative increment and RANTES/CCL5 and TNF-α show relative high level at day 0 of storage, moreover, no obvious increase of accumulation is observed along with prolonging of the storage time, suggesting no relation of concentration with storage time.
Adult ; Blood Donors ; Blood Preservation ; Chemokine CCL2 ; blood ; Chemokine CCL5 ; blood ; Erythrocytes ; metabolism ; Humans ; Male ; Platelet-Derived Growth Factor ; metabolism ; Tumor Necrosis Factor-alpha ; blood ; Vascular Endothelial Growth Factor A ; blood ; Young Adult

OBJECTIVETo investigate the effects of non-neuronal muscarinic receptors (NNMR) stimulation on atherosclerosis and endothelial cells activation.

METHODSAtherosclerosis model was established in ApoE-/- mice by a high fat diet for 7 weeks. During the experimental periods, animals were received a low (7 mg/kg/d) or a high (21 mg/kg/d) dose of arecoline by gavage. At the termination of the treatments, serum total cholesterol and NO levels were measured, and the aorta morphology was analyzed by hematoxylin and eosin staining. The gene expression of monocyte chemoattractant protein-1 (MCP-1) and adhesion molecules in the thoracic aortas was determined by RT-PCR, and the MCP-1 protein expression and NF-κB activity were detected by Western blot analysis. NO production, MCP-1 secretion in cultured rat aortic endothelial cells (RAECs), and monocyte-endothelium adhesion assay were also performed after arecoline treatments.

RESULTSArecoline efficiently decreased atherosclerotic plaque areas, increased serum nitric oxide (NO) content, suppressed the mRNA and protein expression of MCP-1, and modulated the IκB-α degradation and P65 phosphorylation in the aortae of ApoE-/- mice. Furthermore, arecoline promoted NO production and suppressed MCP-1 secretion in cultured RAECs after ox-LDL exposure, and either atropine or NG-nitro-L-arginine methylester could abrogate these effects. Arecoline also significantly inhibited the adherence of U937 monocytes to the ox-LDL injured human umbilical vein endothelial cells, which could be abolished by atropine.

CONCLUSIONOur results indicate that arecoline attenuates the progression of atherosclerosis and inhibits endothelial cells activation and adherence by stimulating endothelial NNMR. These effects, at least in part, are due to its modulation on NF-κB activity.

Animals ; Aorta ; cytology ; Apolipoproteins E ; Arecoline ; pharmacology ; Atherosclerosis ; physiopathology ; prevention & control ; Cell Adhesion Molecules ; metabolism ; Chemokine CCL2 ; metabolism ; Cholesterol ; blood ; Disease Progression ; Endothelial Cells ; cytology ; drug effects ; Endothelium, Vascular ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; I-kappa B Proteins ; metabolism ; Lipoproteins, LDL ; Mice ; Mice, Knockout ; Monocytes ; cytology ; NF-KappaB Inhibitor alpha ; Nitric Oxide ; blood ; Nitroarginine ; pharmacology ; Rats ; Receptors, Muscarinic ; physiology ; Transcription Factor RelA ; metabolism

In traditional Chinese medicine, Nitraria sibirica Pall. (Nitrariaceae) is used to treat hypertension. This study determined the effects of the total alkaloids of the leaves of Nitraria sibirica (NSTA) on blood pressure and albuminuria in mice treated with angiotensin II and a high-salt diet (ANG/HS). Adult mice were divided into three groups: control; infused with angiotensin II and fed a diet containing 4% NaCl (ANG/HS; and ANG/HS plus injection of NSTA (1 mg·kg(-1)·d(-1), i.p.). After treatment of these regimens, daily water and food intake, kidney weight, blood pressure, urinary albumin excretion, renal concentrations of inflammatory markers, including soluble intercellular adhesion molecule-1 (sICAM-1) and monocyte chemoattractant protein-1 (MCP-1), and the expression of renal fibrosis markers were determined. Compared to the control group, the ANG/HS group had higher blood pressure and urinary albumin excretion. Treatment with NSTA in ANG/HS mice for three weeks significantly reduced blood pressure and urinary albumin excretion. ANG/HS treatment caused elevated levels of sICAM-1 and MCP-1, as well as increased fibrosis markers. Concurrent treatment with ANG/HS and NSTA attenuated the levels and expression of renal inflammatory and fibrosis markers. Treatment with NSTA effectively reduces hypertension-induced albuminuria through the reduction of renal inflammatory and fibrosis markers.
Albuminuria ; drug therapy ; metabolism ; physiopathology ; Alkaloids ; administration & dosage ; Angiotensin II ; metabolism ; Animals ; Blood Pressure ; drug effects ; Chemokine CCL2 ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Hypertension ; drug therapy ; metabolism ; physiopathology ; Magnoliopsida ; chemistry ; Male ; Mice ; Rats, Sprague-Dawley ; Sodium Chloride, Dietary ; adverse effects ; metabolism

OBJECTIVE: To investigate the effects of serum from the obesity patients and obesity patients with Diabetic mellitus on toll-like receptor 4/Nuclear factor -κB p65 (TLR/NF-κB) pathway in human THP-1 monocytes and to explore the inflammatory immune response in obesity. METHODS: Peripheral serum was isolated from healthy volunteers (the control group), the obesity patients (Ob group) and the obesity patients with diabetic mellitus (the Ob with DM group), respectively, 20 in each group. THP-1 monocytes were incubated with the serum for 48 h. The monocytes and culture supernatant were collected. The phosphorylation level of NF-κB p65 protein in THP-1 monocytes was evaluated by Western blot as well as immunofluorescence assay. The TLR4 mRNA expression was evaluated by RT-PCR. ELISA was used to measure the monocyte chemotactic protein-1 (MCP-1) levels in the culture supernatant. RESULTS: In the presence of serum, the obesity group and the obesity with diabetic mellitus group showed the up-regulated phosphorylation level of NF-κB p65 protein and TLR4 mRNA expression in THP-1 monocytes compared with the healthy control group (both P<0.05), and the MCP-1 levels in the obesity patients were up-regulated significantly compared with the healthy control group [healthy control group (26.4 ± 3.9) pg/mL, Ob group (45.8 ± 10.0) pg/mL, Ob with DM group (58.0 ± 15.3) pg/mL; P<0.05]. These parameters were further up-regulated in the obesity patients with diabetic mellitus patients. CONCLUSION The serum from the obesity patients or the obesity patients with diabetes can induce monocyte dysfunction, which might be related to the activation of TLR4/NF-κB signaling pathway.
Cell Line ; Chemokine CCL2 ; Diabetes Mellitus ; blood ; Humans ; Monocytes ; cytology ; metabolism ; Obesity ; blood ; Phosphorylation ; Serum ; Signal Transduction ; Toll-Like Receptor 4 ; metabolism ; Transcription Factor RelA ; metabolism ; Up-Regulation

OBJECTIVETo observe the effect of Huanglian Jiedu IJecoction (HJU) on systemic and vascular immune responses of high fat diet fed apoE deficient (apoE(-/-)) mice.

METHODSEight wild type C57BL6 mice were recruited as the wild type common food group. Totally 24 apoE(-/-) mice were randomly divided into the ApoE'common food group, the ApoE(-/-) hyperlipidemia group, and the ApoE(-/-) hyperlipidemia plus HJD group, 8 in each group. In the present study, the common food mice and high fat fed mice were fed with a chow diet or a high cholesterol diet for 4 weeks. HJD was given to mice in the ApoE(-/-) hyperlipidemia plus HJD group at the daily dose of 5 g/kg by gastrogavage, while equal volume of pure water was given to mice in the rest groups by gastrogavage. Four weeks later, the plasma levels of blood lipids, the ratio of peripheral blood mononuclear cells, and expressions of Toll-like receptor 4 (TLR-4) and CD36 on the monocytes were detected. The pathological changes and expressions of cytokines in local aorta were detected. The plasma cytokine levels in response to lipopolysaccharide (LPS) were analyzed. Results (1) Compared with the wild type common food group, TO, TG, and LDL-O significantly increased in the ApoE(-/-) common food group (P < 0. 05, P < 0.01). Compared with the ApoE(-/-) common food group, TC and LDL-C significantly increased in the hyperlipidemia group (P < 0. 05). There was no statistical difference in each index between the ApoE(-/-) hyperlipidemia group and the ApoE(-/-) hyperlipidemia plus HJD group (P > 0.05). (2) Compared with the wild type common food group, no obvious change of the ratio of peripheral blood mononuclear cells happened, the TLR4 expression level significantly increased in the ApoE'common food group (P < 0. 05). Compared with the ApoE common food group, the ratio of peripheral blood mononuclear cells and the TLR4 expression level significantly increased in the ApoE' hyperlipidemia group (P < 0.05). Compared with the ApoE(-/-) hyperlipidemia group, the ratio of peripheral blood mononuclear cells and the TLR4 expression level significantly decreased. Besides, the CD36 expression level also significantly decreased (P<0.05). (3) After stimulated by LPS for 3 h, compared with the wild type common food group, plasma TNF-ct and IL-b expressions significantly increased in the ApoE(-/-) common food group (P < 0.05). Compared with the ApoE(-/-) common food group, plasma expressions of IL-12, TNF-alpha, MCP-1, and IL-10 increased, but with no statistical difference in the ApoE(-/-) hyperlipidemia group (P > 0.05). After 4-week intervention of HJD, compared with the ApoE(-/-) hyperlipidemia group, the MCP-1 expression was significantly down-regulated, while the IL-10 expression significantly increased, showing statistical difference (P < 0.05). Compared with the wild type common food group, mRNA expression levels of IFN-gamma, MCP-1 , TNF-alpha, IL-10, and IL-1beta significantly increased (P < 0. 05, P < 0.01). Compared with the ApoE(-/-) common food group, not only mRNA expression levels of IFN-gamma, MCP-1, TNF-alpha, and IL-1beta, further significantly increased, but also IL-12, IL-10, and TGF-beta significantly increased (P < 0. 05, P < 0. 01). After 4-week intervention of HJD, compared with the ApoE(-/-) hyperlipidemia group, mRNA expression levels of MCP-1, TNF-alpha, IL-1beta, and IL-12 significantly decreased in the ApoE(-/-) hyperlipidemia plus HJD group (P < 0.05, P < 0.01).

CONCLUSIONSHigh fat diet induced systemic reaction and inflammatory reactions of local vessels. The local inflammatory response of vessels exceeded systemic inflammatory response. Intervention of HJD could attenuate inflammatory response, especially in local arteries. Meanwhile, it enhanced systemic anti-inflammatory reactions.

Animals ; Aorta ; pathology ; Apolipoproteins E ; genetics ; CD36 Antigens ; metabolism ; Chemokine CCL2 ; metabolism ; Dietary Fats ; adverse effects ; Drugs, Chinese Herbal ; pharmacology ; Female ; Hyperlipidemias ; blood ; etiology ; immunology ; Inflammation ; Interleukin-10 ; blood ; Interleukin-12 ; blood ; Interleukin-1beta ; blood ; Leukocytes, Mononuclear ; metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Systemic Inflammatory Response Syndrome ; blood ; etiology ; immunology ; Toll-Like Receptor 4 ; metabolism ; Transforming Growth Factor beta ; blood ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo observe the effect of electroacupuncture (EA) at Fenglong (ST40) on inflammatory factors in macrophages of hyperlipemia (HLP) rats, and to explore the role of EA in treating HLP.

METHODSTotally 50 adult male SD rats were selected. After one-week adaptative feeding, they were randomly divided into 5 groups, i.e., the blank control group (Group A), the model group (Group B), the diet control group (Group C), the EA group (Group D), the diet control + EA group (Group E), 10 in each group. By the end of treatment, all rats were sacrificed to get blood from the carotid artery. The contents of serum total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), and low density lipoprotein cholesterol (LDL-C) were determined. Peritoneal macrophages were isolated. Contents of intercellular adhesion molecule-1 (ICAM-1), monocyte chemoattractant protein 1 (MCP-1), tumor necrosis factor-alpha (TNF-alpha), and interleukin 6 (IL-6) were detected using flow cytometry (FCM) after adding the fluorescein-labeled molecule antibodies of inflammatory factors.

RESULTSCompared with Group A, the serum contents of TC and LDL-C obviously increased and HDL-C significantly decreased in Group B (all P < 0.01). There was no obvious change in the content of TG between Group A and Group B (P > 0.05). Compared with Group B, the contents of TC and LDL-C significantly decreased in Group C, D, and E (P < 0.01). Compared with Group C, serum contents of TC and LDL-C obviously decreased in Group D and E (P < 0.01). Compared with Group B, the content of macrophages significantly increased in Group A (P < 0.01). Compared with Group B, the content of macrophages significantly decreased in Group D and E (P < 0.01), more obviously in Group E (P < 0.01). Compared with Group C, the content of macrophages significantly decreased in Group E (P < 0.05). Compared with Group A, levels of ICAM-1, MCP-1, TNF-alpha, and IL-6 in the peritoneal macrophages significantly increased in Group B (P < 0.01). Compared with Group B, levels of ICAM-1, MCP-1, TNF-alpha, and IL-6 significantly decreased in Group C, D, and E, but more obviously in Group E (P < 0.01). Compared with Group C, levels of ICAM-1, MCP-1, TNF-alpha (P < 0.05, P < 0.01), and IL-6 (P < 0.05) significantly decreased in Group E.

CONCLUSIONEA at Fenglong (ST40) could obviously downregulate the serum levels of TC and LDL-C in HLP rats, reduce the contents of macrophages, ICAM-1, MCP-1, TNF-alpha, and IL-6, thus arriving at the therapeutic effect in treating HLP.

Acupuncture Points ; Animals ; Chemokine CCL2 ; metabolism ; Cholesterol ; blood ; Cholesterol, LDL ; blood ; Electroacupuncture ; Hyperlipidemias ; metabolism ; therapy ; Intercellular Adhesion Molecule-1 ; metabolism ; Macrophages ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism

BACKGROUNDMycophenolate mofetil (MMF) has been used to prevent transplant rejection for many years and has been shown to have protective effects against renal failure. The objective was to investigate the effect of MMF on monocyte Toll-like receptor 4 (TLR4) signaling in the early stages of renal ischemia/reperfusion injury (IRI) of mice.

METHODSSixty BALB/C mice were randomly divided into two groups: an IRI group, in which renal IRI was induced by clamping the renal pedicles for 45 minutes, and an MMF group, in which MMF was given (40 mg×kg(-1)×d(-1), intraperitoneally) from 2 days before renal IRI. The plasma creatinine level and renal tissue damage of each group mice were observed 6, 12, 24, and 48 hours after reperfusion. The concentration of plasma high-mobility group box 1 (HMGB-1) (TLR4 ligand), interleukin 6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), and tumor necrosis factor α (TNF-α) and the expression of TLR-4 on monocytes were determined.

RESULTSThe plasma creatinine concentration in the MMF group was lower compared to the IRI group (after reperfusion of 6, 12, 24, or 48 hours, P < 0.05). Pathological analysis showed that the renal damage was slighter, TLR-4 expression was reduced (after reperfusion of 6, 12, 24, or 48 hours, P < 0.05), and the concentration of cytokines in the plasma was lower (P < 0.05) in the MMF group. No differences in the concentrations of HMGB-1 were observed (P > 0.05).

CONCLUSIONMonocyte TLR4 signaling is important in the early stage of kidney IRI, but MMF can inhibit it and improve renal function.

Acute Kidney Injury ; blood ; drug therapy ; metabolism ; Animals ; Chemokine CCL2 ; genetics ; metabolism ; Creatinine ; blood ; Cytokines ; blood ; HMGB1 Protein ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred BALB C ; Monocytes ; drug effects ; metabolism ; Mycophenolic Acid ; analogs & derivatives ; therapeutic use ; Random Allocation ; Reperfusion Injury ; blood ; drug therapy ; metabolism ; Signal Transduction ; drug effects ; genetics ; Toll-Like Receptor 4 ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism