AIMTo investigate the effect of NOS and PTEN on the hypertrophic response induced by angiotensin II in the primary culture of neonatal rat cardiomyocytes.

METHODSTotal protein content of cardiomyocytes was used as the index of cardiac myocyte hypertrophy. eNOS mRNA, iNOS mRNA and PTEN mRNA expression were assessed using RT-PCR normalized with GAPDH. PTEN protein was determined by Western blot and immunohistochemistry method.

RESULTS(1) On day 1 after Ang II treatment, the expression of eNOS mRNA was significantly decreased whereas iNOS mRNA expression was significantly increased. The effect of Ang II on NOS expression was inhibited by L-arginine. (2) Total protein content of cardiomyocytes increased significantly on day 5 after Ang II treatment, and PTEN protein expression was significantly decreased. The increased protein content and the decreased expression of PTEN protein were inhibited by L-Arg. The L-arginine effect was blocked by L-NAME(NOS inhibitor). (3) The positive immunocytochemical product of PTEN was mainly located in the nucleus of myocardiocyte.

CONCLUSIONThese results indicate that NOS and PTEN may take part in the process of cardiac myocyte hypertrophy induced by Ang II. The effect of L-arginine on cardiomyocytes may be mediated by NOS/NO and PTEN.

Angiotensin II ; pharmacology ; Animals ; Animals, Newborn ; Cardiomegaly ; chemically induced ; metabolism ; pathology ; Cells, Cultured ; Myocytes, Cardiac ; drug effects ; metabolism ; pathology ; Nitric Oxide Synthase Type II ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; PTEN Phosphohydrolase ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the role of tumor suppressor PTEN in cardiac hypertrophy, the expression of PTEN mRNA and protein was analyzed in the tissue of left ventricle in abdominal aorta constricted-induced cardiac hypertrophic rats which treated with and without captopril. The expression of PTEN mRNA and protein in cultured neonatal rat cardiomyocyte treated with AngII was studied.

METHODSSD rats were divided into control group, hypertrophy group and captopril group. The expression of PTEN in different groups at 2 and 4 weeks after operation as well as in cultured neonatal rat cardiomyocyte treated with AngII was detected by RT-PCR and Western blot. The localization of PTEN in left ventricle and cultured cardiomyocyte was determined by immunohistochemistry.

RESULTS(1) Compared with control group, the expressions of PTEN mRNA and protein in left ventricle of hypertrophy group as well as in cultured cardiomyocyte treated with AngII were reduced. (2) Compared with hypertrophy group, the expressions of PTEN mRNA and protein in left ventricle of captopril group were upregulated, which were similar to those of control group. (3) Positive immunohistochemical staining of PTEN was located in the nucleus of cardiomyocytes.

CONCLUSIONPTEN may play a negative regulation role in the process of cardiac hypertrophy, and the role of PTEN may be closely related with renin-angiotensin system.

Animals ; Captopril ; metabolism ; Cardiomegaly ; metabolism ; pathology ; Gene Expression ; Genes, Tumor Suppressor ; Male ; Myocytes, Cardiac ; metabolism ; PTEN Phosphohydrolase ; metabolism ; Rats ; Rats, Sprague-Dawley

AIMTo investigate the role of tumor suppressor PTEN in cardiac hypertrophy, the expression of PTEN mRNA in left ventricle of abdominal aorta constricted-induced cardiac hypertrophic rats which treated with and without captopril was analyzed.

METHODSSD rats were divided into control group, hypertrophy group and captopril group. The expression of PTEN mRNA in left ventricle was detected by RT-PCR in different groups in 4 weeks after operation.

RESULTS(1) Compared with control group, the expression of PTEN mRNA in left ventricle of hypertrophy group was reduced. (2) Compared with hypertrophy group, the expression of PTEN mRNA in left ventricle of captopril group was upregulated, which were similar to that of control group.

CONCLUSIONPTEN maybe plays a negative regulation role in the process of cardiac hypertrophy, and the role of PTEN is closely relative with renin-angiotensin system.

Angiotensin-Converting Enzyme Inhibitors ; pharmacology ; Animals ; Captopril ; pharmacology ; Cardiomegaly ; metabolism ; pathology ; Male ; PTEN Phosphohydrolase ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

AIMTo investigate the effect of Naoyikang serum on the damage induced by glutamate in hippocampal neuron.

METHODSMorphological observation, MTT assay and nuclear DNA-associated fluorescence with DAPI dye were applied to evaluate the viability of hippocampal neuron, immunocytochemistry and RT-PCR were used to determine the expression of PTEN.

RESULTSA decreased viability and increased expression of PTEN were shown in hippocampal neuron in response to the treatment with glutamate. It was shown that the percentage of cell death and the expression of PTEN were reduced by the treatment with Naoyikang serum.

CONCLUSIONThese results suggest that Naoyikang may prevent the toxicity of glutamate by suppressing the expression of PTEN.

Animals ; Animals, Newborn ; Cell Death ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Glutamic Acid ; pharmacology ; Hippocampus ; cytology ; drug effects ; metabolism ; Neurons ; drug effects ; metabolism ; Neuroprotective Agents ; pharmacology ; PTEN Phosphohydrolase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Serum

OBJECTIVETo investigate weather there is a toll-like receptor 4 (TLR4)-mediated myeloid differentiation factor 88 (MyD88)-dependent pathway in hippocampal neurons of rats and the probable role of the pathway in neuroinflammation.

METHODSTo establish the proper model, primarily cultured hippocampal neurons were treated with lipopolysaccharides (LPS), or pretreated with TLR4 antibody then co-treated with LPS. The expression of mRNA of MyD88 and TNF-alpha receptor associated factor 6 (TRAF6) were tested by RT-qPCR. The content of MyD88 and TRAF6 were tested by Western blot. The nuclear translocation of nuclear factor-kappaB/P65 (NF-kappaB/p65) was tested by immunofluorescence. The content of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta) and nitric oxide (NO) were tested by ELISA.

RESULTSLPS could increase MyD88 and TRAF6 mRNA, upregulate protein level of MyD88 and TRAF6 and increase the level of TNF-alpha, IL-1beta and NO in cell culture supernatant. LPS also could promote NF-kappa B/p65 translation to the nucleus. The pretreatment with TLR4 antibody reduced the translocation to nucleus for NF-kappaB/P65 and the contents of TNF-alpha, IL-1beta and NO in the culture supernatant.

CONCLUSIONThere is a TLR4-mediated MyD88-dependent pathway in hippocampal neurons. The activation of this pathway can increase the level of TNF-alpha, IL-1beta and NO in cell culture supernatant. TLR4-mediated MyD88-dependent pathway in hippocampal neurons participate in neuroinflammation, that means neurons are not passive in inflammation.

Animals ; Cells, Cultured ; Hippocampus ; cytology ; metabolism ; Interleukin-1beta ; metabolism ; Myeloid Differentiation Factor 88 ; metabolism ; Neuritis ; metabolism ; Neurons ; metabolism ; Nitric Oxide ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; TNF Receptor-Associated Factor 6 ; metabolism ; Toll-Like Receptor 4 ; metabolism ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate the effect and mechanism of meloxicam on the inflammatory reaction induced by beta amyloid protein (AB) in Alzheimer's disease (AD) rats.

METHODSThe rat model was established by microinjection of Abeta(1-40) into hippocampus. The expression of NF-kappaB p65 and glial fibrillary acidic protein (GFAP) in hippocampus were detected by immunohistochemistry. The content of GFAP in cortex was tested by Western-blot. The content of TNF-alpha in cortex was tested by ELISA. The expression of IL-1beta mRNA was tested by RT-PCR.

RESULTSThe expression of NF-kappaB p65, GFAP and TNF-alpha as well as IL-1beta mRNA were decreased by meloxicam.

CONCLUSIONMeloxicam can reduce the proliferation of astrocyte by decreasing the expression of GFAP in AD model rat's hippocampus and cortex. And the depression of NF-kappaB p65 may significantly decrease the expression of TNF-alpha1 and IL-1beta to lessen the inflammatory reaction in cerebral tissue.

Alzheimer Disease ; chemically induced ; drug therapy ; pathology ; Amyloid beta-Peptides ; toxicity ; Animals ; Cerebral Cortex ; metabolism ; pathology ; Glial Fibrillary Acidic Protein ; metabolism ; Inflammation ; prevention & control ; Interleukin-1beta ; metabolism ; Male ; Peptide Fragments ; toxicity ; Rats ; Rats, Sprague-Dawley ; Thiazines ; pharmacology ; therapeutic use ; Thiazoles ; pharmacology ; therapeutic use ; Transcription Factor RelA ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate the effect of acupoint injection of oxymatrine (OM) on experimental hepatocellular carcinoma and the mechanism.

METHODSThe rats of hepatocellular carcinoma induced by 2-acetoaminoflurence (2-AAF) were randomly divided into a normal control group (group N), a model group (group M), a control group of oxymatrine intraperitoneal injection (OM ip group) and a treatment group of small dose oxymatrine injection into Zusanli (OM ZSL group). At the end of 12h week, the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma-glutamyl transferase (gamma-GT) were determined. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expressions of cyclin D1 and cyclin-dependent kinase 4 (CDK4) mRNA in hepatocellular carcinoma tissues.

RESULTSThe number of cancer nodes on the surface of liver in th Om ip group and the Om ZSL group was lower than in the group M, with the serum ALT, AST, and gamma-GT levels significantly decreased (P<0. 01), and significantly inhibited expressions of cyclin D1, CDK4 mRNA (P<0. 01).

CONCLUSIONOM ip and small dose oxymatrine injection into ZSL can treat or delay hepatocarcinogenisis of hepatocellular carcinoma induced by 2-AAF. Partial mechanism of this anti-carcinoma is protecting hepatocytes possibly through improving hepatic functions, and inhibiting excessive proliferation of liver cancer cells via inhibiting the expressions of cyclin Dl, CDK4 mRNA.

Acupuncture Points ; Alanine Transaminase ; blood ; Alkaloids ; administration & dosage ; Animals ; Aspartate Aminotransferases ; blood ; Cyclin D1 ; genetics ; Cyclin-Dependent Kinase 4 ; genetics ; Injections ; Liver Neoplasms, Experimental ; drug therapy ; Male ; Quinolizines ; administration & dosage ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; gamma-Glutamyltransferase ; blood

AIMTo investigate the mechanisms of Naoyikang (Traditional Chinese Medicine) on the Alzheimer's Disease (AD) model mice induced by D-galactose (D-gal) and NaNO2.

METHODSThe mouse model was established by intraperitoneal injection of D-gal and NaNO2. The capacity of learning and memory was tested on mice with electrical maze; the content of nitric oxide (NO) and the activity of monoamine oxidase-B (MAO-B), glutathione peroxidase (GSH-PX), Na(+) -K(+) -ATP enzyme and Ca(2+) -ATP enzyme in cerebral cortex and hippocampus were assayed by biochemical methods; expression of Bax and Bcl-2 mRNA was detested by RT-PCR.

RESULTSNaoyikang could ameliorate the capacity of learning and memory of AD model mice and reduce MAO-B activity in the brain tissue and activate the activity of Na(+) -K(+) -ATP enzyme and Ca(2+) -ATP enzyme in the brain tissue and decrease the expression of Bax mRNA, but increase the expression of Bcl-2 mRNA in the model brain tissue.

CONCLUSIONNaoyikang could protect AD model mice induced by D-gal and NaNO2. It could modify the metabolism of monoamine neurotransmitter in brain through reducing MAO-B activity and protect neurons by activating the activity of Na(+) -K(+) -ATP enzyme and Ca(2+) -ATP enzyme and decrease Bax expression and increase Bcl-2 expression in the model brain tissue.

Alzheimer Disease ; chemically induced ; drug therapy ; Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Galactose ; Male ; Maze Learning ; Mice ; Mice, Inbred ICR ; Neuroprotective Agents ; therapeutic use ; Phytotherapy ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Sodium Nitrite ; Sodium-Potassium-Exchanging ATPase ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

AIM:To investigate the therapeutic effect of oxymatrine on non-small-cell lung cancer(NSCLC) A549 cells and a xenograft mouse model,and to explore the underlying molecular mechanisms. METHODS:The effect of oxymatrine on the A549 cell viability was assessed by CCK-8 assay. After the A549 cells were treated with Toll-like re?ceptor 4(TLR4)stimulator lipopolysaccharide(LPS)and oxymatrine(5,10 and 15 mmol/L),the mRNA and protein ex?pression levels of TLR4 and myeloid differentiation factor 88(MyD88)were analyzed by RT-qPCR and Western blot,re?spectively. The migration and invasion abilities of the cells were measured by Transwell assay,and the mRNA and protein expression levels of matrix metalloproteinases-2(MMP-2),MMP-9 and vascular endothelial growth factor(VEGF)were also determined. A xenograft model in nude mice was utilized to evaluate the effect of oxymatrine on tumor growth. RE?SULTS:Oxymatrine inhibited the viability of A549 cells,decreased LPS-induced expression of TLR4,MyD88,MMP-2, MMP-9 and VEGF in A549 cells,and suppressed LPS-increased migration and invasion abilities of A549 cells. In the xe?nograft model,oxymatrine both reduced tumor growth and inhibited TLR4 expression in the tumor. CONCLUSION:Oxy?matrine exerts anti-tumor properties in NSCLC in vitro and in vivo by down-regulating the TLR4/MyD88 signaling pathway, suggesting that oxymatrine can be a potential therapeutic agent for NSCLC.

OBJECTIVETo investigate foot biomechanics characteristic of patients with type 2 diabetes mellitus.

METHODSThis study was conducted among 303 patients with type 2 diabetes. The whole foot was divided into 10 regions, namely the first toe (T1); the second to fifth toes (T2-5); the first, second, third, fourth, and fifth metatarsals (M1, M2, M3, M4, and M5, respectively); midfoot (MF), and the heel medial (HM). Foot arch index, foot angle and maximum peak pressure (MPP) of the 10 regions were measured using a Footscan gait system.

RESULTSThe maximum peak pressure of 10 regions decreased in the order of M3>M2>HM>M4>HL>M1>M5>T1>ML>T2-5 for the left foot, and in the order of M3>M2>HM>M4>HL>M1>M5>T1>ML>T2-5 for the right foot. The MPP in M1 region was higher in the right than in the left foot (P<0.05). The MPP in M3, M4, M5, and MF was higher in the left than in the right foot (P<0.05). The percentage of high-risk foot (defined by a total plantar pressure ≥70 N/cm) was 34% on the left and 17.7% on the right. An increased BMI was associated with a significant increase in high-risk foot, but not for the right foot in underweight patients. Foot flat phase was extended and forefoot push-off phase shortened in stance phase in the patients. Compared with the right foot, the left foot showed a significantly increased foot arch index and increased low and high arch rates with a decreased normal arch rate. Total plantar pressure was higher in of the left high arch foot than in normal arch foot. The foot angle was significantly larger on the right than on the left. The bilateral total plantar pressures were significantly greater in male patients (P<0.05) and increased with age but were not associated with the duration of DM, foot angle, or glycosylated hemoglobin level.

CONCLUSIONDiabetic patients have obvious alterations in foot biomechanics with abnormalities of the plantar pressure, and the percentage of high-risk foot increases in overweight and obese patients, suggesting the need of body weight control in these patients when administering offloading treatment for prevention of diabetic foot ulcer.

Biomechanical Phenomena ; Diabetes Mellitus, Type 2 ; physiopathology ; Diabetic Foot ; prevention & control ; Female ; Foot ; physiopathology ; Gait ; Heel ; physiopathology ; Humans ; Male ; Obesity ; physiopathology ; Overweight ; physiopathology ; Pressure