This study aimed to investigate the effect and mechanism of Zuogui Jiangtang Qinggan Formula(ZGJTQG) on the glucolipid metabolism of type 2 diabetes mellitus(T2DM) complicated with non-alcoholic fatty liver disease(NAFLD). NAFLD was induced by a high-fat diet(HFD) in MKR mice(T2DM mice), and a model of T2DM combined with NAFLD was established. Forty mice were randomly divided into a model group, a metformin group(0.067 g·kg~(-1)), and high-and low-dose ZGJTQG groups(29.64 and 14.82 g·kg~(-1)), with 10 mice in each group. Ten FVB mice of the same age were assigned to the normal group. Serum and liver tissue specimens were collected from mice except for those in the normal and model groups after four weeks of drug administration by gavage, and fasting blood glucose(FBG) and fasting insulin(FINS) levels were measured. The levels of total cholesterol(TC), triglyceride(TG), and low-density lipoprotein(LDL) were detected by the single reagent GPO-PAP method. Very low-density lipoprotein(VLDL) was detected by enzyme-linked immunosorbent assay(ELISA). Alanine aminotransferase(ALT) and aspartate ami-notransferase(AST) were determined by the Reitman-Frankel assay. The pathological changes in the liver were observed by hematoxylin-eosin(HE) staining and oil red O staining. Real-time fluorescence-based quantitative polymerase chain reaction(real-time PCR) and Western blot were adopted to detect the mRNA and protein expression of forkhead transcription factor O1(FoxO1), microsomal triglyceride transfer protein(MTP), and apolipoprotein B(APOB) in the liver. The results showed that high-dose ZGJTQG could signi-ficantly reduce the FBG and FINS levels(P<0.05, P<0.01), improve glucose tolerance and insulin resistance(P<0.05, P<0.01), alleviate the liver damage caused by HFD which was reflected in improving liver steatosis, and reduce the serum levels of TC, TG, LDL, VLDL, ALT, and AST(P<0.05, P<0.01) in T2DM mice combined with NAFLD. The findings also revealed that the mRNA and protein expression of FoxO1, MTP, and APOB in the liver was significantly down-regulated after the intervention of high-dose ZGJTQG(P<0.05, P<0.01). The above study showed that ZGJTQG could effectively improve glucolipid metabolism in T2DM combined with NAFLD, and the mechanism was closely related to the regulation of the FoxO1/MTP/APOB signaling pathway.
Mice ; Animals ; Non-alcoholic Fatty Liver Disease/metabolism* ; Diabetes Mellitus, Type 2/metabolism* ; Liver ; Lipoproteins, LDL/metabolism* ; Signal Transduction ; Diet, High-Fat/adverse effects* ; RNA, Messenger/metabolism*

The saponins in different parts of Gynostemma pentaphyllum were analyzed via UPLC-Q-TOF-MS~E. A total of 46 saponins were identified, and the underground part had 26 saponins more than the aboveground part, most of which were trisaccharide saponins. The rat model of hyperlipidemia was established with high-fat diet. This study explored the lipid-lowering activity of total saponins in the underground part of G. pentaphyllum, so as to provide a theoretical basis for the comprehensive utilization of the underground part of G. pentaphyllum. A total of 99 healthy SD rats were randomly assigned into a blank group, a model group, a positive drug group, an aboveground total saponins group, and low-, medium-, and high-dose underground total saponins groups. Except the blank group, the other groups were fed with high-fat diet for 6 weeks. Then, the blood was collected from the orbital cavity to determine whether the modeling was successful according to the serum levels of total cholesterol(TC) and triglyceride(TG). After intragastric administration of the corresponding agents for 30 continuous days, the physical state of the rats were observed, and the body weight and liver specific gravity were measured. Furthermore, the levels of TC, TG, low-density lipoprotein cholesterol(LDL-C), high-density lipoprotein cholesterol(HDL-C), alanine transaminase(ALT), aspartate transaminase(AST), bilirubin, and total bile acids in serum, as well as the levels of superoxide dismutase(SOD), malondialdehyde(MDA), peroxidase proliferator-activated receptor(PPAR-γ) in the liver tissue, were determined. The pathological changes of liver was observed via HE staining. The results showed that the aboveground total saponins and medium-and high-dose underground total saponins can treat hepatocyte steatosis, lower TC, TG, LDL-C, ALT, AST, total bilirubin, MDA, and PPAR-γ levels, and increase HDL-C and SOD levels in the model rats. The effect tended to be more obvious with the increase in dosage. Therefore, the total saponins in the underground part of G. pentaphyllum have good pharmacological effect of reducing blood lipid, which provides a theoretical basis for the comprehensive utilization of the underground part of G. pentaphyllum.
Alanine Transaminase/analysis* ; Animals ; Aspartate Aminotransferases/analysis* ; Bile Acids and Salts/blood* ; Bilirubin/blood* ; Cholesterol, LDL/blood* ; Diet, High-Fat/adverse effects* ; Gynostemma/chemistry* ; Hypolipidemic Agents/therapeutic use* ; Lipoproteins, HDL/blood* ; Liver/metabolism* ; Malondialdehyde/analysis* ; Peroxisome Proliferator-Activated Receptors/analysis* ; Rats ; Rats, Sprague-Dawley ; Saponins/therapeutic use* ; Superoxide Dismutase ; Triglycerides/blood* ; Trisaccharides/therapeutic use*

Apoptosis ; Human Umbilical Vein Endothelial Cells ; Humans ; Lipoproteins, LDL/adverse effects* ; MicroRNAs/genetics* ; Molecular Chaperones/genetics*

Cardio-cerebral vascular disease induced by atherosclerosis is a serious cause of human health. The pathogenesis of AS is very complex,and the oxidized low-density lipoprotein( ox LDL) induced foam cells formation is considered to be the most important cytological change in AS. Based on the definition of " TCM chemical biology",we clarified the chemical composition of Ilex hainanensis,the effective substances of I. hainanensis on the activity of anti-AS were screened. Then we found that saponin BF523 had the good inhibitory effect on foam cell formation. In this research,we studied the BF523 as the research object to clarify the molecular target of the active compound of I. hainanensis by foam cell formation model. The results showed that BF523 significantly inhibited the oxidation of ox LDL-induced macrophage foaming and decreased the lipid content in macrophages. BF523 had inhibited the phagocytosis of ox LDL in macrophages by reducing the mRNA and protein levels of scavenger receptor CD36,thereby inhibiting the occurrence and development of AS. These findings not only clarified the mechanism of the inhibition of foam cell formation by saponin BF523,but also provided a useful exploration for the enrichment of the theory of " TCM chemical biology".
Atherosclerosis ; CD36 Antigens ; metabolism ; Cells, Cultured ; Foam Cells ; cytology ; drug effects ; Humans ; Ilex ; chemistry ; Lipoproteins, LDL ; adverse effects

OBJECTIVETo investigate the role of aqueous extracts of Tribulus terrestris (TT) against oxidized low-density lipoprotein (ox-LDL)-induced human umbilical vein endothelial cells (HUVECs) dysfunction in vitro.

METHODSHUVECs were pre-incubated for 60 min with TT (30 and 3 μg/mL respectively) or 10(-5) mol/L valsartan (as positive controls) and then the injured endothelium model was established by applying 100 μg/mL ox-LDL for 24 h. Cell viability of HUVECs was observed by real-time cell electronic sensing assay and apoptosis rate by Annexin V/PI staining. The cell migration assay was performed with a transwell insert system. Cytoskeleton remodeling was observed by immunofluorescence assay. The content of endothelial nitric oxide synthase (eNOS) was measured by enzyme-linked immunosorbent assay. Intracellular reactive oxygen species (ROS) generation was assessed by immunofluorescence and flow cytometer. Key genes associated with the metabolism of ox-LDL were chosen for quantitative real-time polymerase chain reaction to explore the possible mechanism of TT against oxidized LDL-induced endothelial dysfunction.

RESULTSTT suppressed ox-LDL-induced HUVEC proliferation and apoptosis rates significantly (41.1% and 43.5% after treatment for 3 and 38 h, respectively; P<0.05). It also prolonged the HUVEC survival time and postponed the cell's decaying stage (from the 69th h to over 100 h). According to the immunofluorescence and transwell insert system assay, TT improved the endothelial cytoskeletal network, and vinculin expression and increased cell migration. Additionally, TT regulated of the synthesis of endothelial nitric oxide synthase and generation of intracellular reactive oxygen species (P<0.05). Both 30 and 3 μg/mL TT demonstrated similar efficacy to valsartan. TT normalized the increased mRNA expression of PI3Kα and Socs3. It also decreased mRNA expression of Akt1, AMPKα1, JAK2, LepR and STAT3 induced by ox-LDL. The most notable changes were JAK2, LepR, PI3Kα, Socs3 and STAT3.

CONCLUSIONSTT demonstrated potential lowering lipid benefits, anti-hypertension and endothelial protective effects. It also suggested that the JAK2/STAT3 and/or PI3K/AKT pathway might be a very important pathway which was involved in the pharmacological mechanism of TT as the vascular protective agent.

Apoptosis ; drug effects ; Cell Movement ; drug effects ; Cell Survival ; drug effects ; Cytoskeleton ; drug effects ; metabolism ; Endothelium, Vascular ; drug effects ; pathology ; physiopathology ; Enzyme-Linked Immunosorbent Assay ; Fluorescent Antibody Technique ; Gene Expression Regulation ; drug effects ; Human Umbilical Vein Endothelial Cells ; drug effects ; Humans ; Lipoproteins, LDL ; adverse effects ; Nitric Oxide Synthase Type III ; metabolism ; Plant Extracts ; pharmacology ; Protective Agents ; pharmacology ; Reactive Oxygen Species ; metabolism ; Tribulus ; chemistry ; Vinculin ; metabolism ; Water ; chemistry

Ginsenoside Rb3 (GRb3) is one of the main components in plasma of Panax quinquefolius Saponin of stem and leaf (PQS), which can be into human plasma. Previous studies have found PQS has estrogen-like vascular protective effects. In the present study, we investigated the estrogen-like protective effect of GRb3 on oxidative stress and dysfunction of endothelial cells induced by oxidized low-density lipoprotein. The activities of SOD, NOS and the contents of MDA in the cell lysate were examined by enzyme method or spectrophotometry. The NO and ET-1 concentrations in the cell culture supernatant were measured by ELISA method. The iNOS and eNOS mRNA expression were measured by real time RT-PCR, while the phosphorylation levels of Akt was measured by Western blotting. The results showed that GRb3 could enhance the activity of SOD, reduce the content of MDA, increase the level of NOS, NO, ET-1 and iNOS mRNA expression while decrease the eNOS mRNA expression and the phosphorylation level of Akt. These effects were blocked by estrogen receptor antagonist ICI182780. GRb3 can play a role in protecting vascular endothelial cells by estrogen receptors, the protective mechanism is similar to 17-β estrodiol.
Cells, Cultured ; Endothelial Cells ; drug effects ; Endothelin-1 ; metabolism ; Estradiol ; analogs & derivatives ; Estrogens ; pharmacology ; Ginsenosides ; pharmacology ; Humans ; Lipoproteins, LDL ; adverse effects ; Nitric Oxide Synthase Type II ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Oxidative Stress ; Panax ; chemistry ; Phosphorylation ; Saponins ; pharmacology ; Superoxide Dismutase ; metabolism

OBJECTIVETo investigate the effects of carbon disulfide (CS(2)) inhalation on the lipid levels of ApoE knockout gene mice and C57BL/6J mice.

METHODSFifty-one male ApoE gene knockout mice were randomly divided into four groups: CS(2)-exposed normal diet group, CS(2)-unexposed normal diet group, CS(2)-exposed high-fat diet group, and CS(2)-unexposed high-fat diet group. Fifty male C57BL/6J mice were divided into four groups in the same way. The exposed groups received 1000 mg/m3 CS(2) by static inhalation (5h/d, 5d/w) for four weeks. The weight of each mouse was determined and recorded once a week. On the 14th day of exposure, six mice in each group were randomly selected to measure serum total cholesterol (TC) levels. On the 28th day of exposure, the serum levels of TC and low-density lipoprotein (LDL) in the remaining mice were measured.

RESULTSThe mean weight gain of exposed groups was less than that of the unexposed groups. On the 14th and 28th days of experiment, the TC levels of the CS2-exposed high-fat diet group were significantly higher than those of the CS(2)-unexposed high-fat diet group among ApoE knockout gene mice (P < 0.01 for both). On the 14th day of experiment, the TC levels of the CS(2)-unexposed high-fat diet group were significantly higher than those of the CS(2)-unexposed normal-diet group among C57BL/6J mice group (P < 0.05). On the 28th day of experiment, the LDL levels of the CS(2)-exposed high-fat diet group were significantly higher than those of the CS(2)-unexposed high-fat diet group among ApoE knockout gene mice (P = 0.003).

CONCLUSIONCS(2) exposure, high-fat diet, and ApoE gene knockout can elevate blood lipids in mice, thus increasing the risk of atherosclerosis.

Administration, Inhalation ; Animals ; Apolipoproteins E ; genetics ; Atherosclerosis ; Body Weight ; Carbon Disulfide ; toxicity ; Diet, High-Fat ; adverse effects ; Gene Knockout Techniques ; Lipid Metabolism ; drug effects ; Lipids ; blood ; Lipoproteins, LDL ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout

BACKGROUNDNon-alcoholic fatty liver disease (NAFLD) is a complex disorder and has been closely linked to obesity. The fat mass and obesity-associated (FTO) gene is a newly discovered gene related to obesity, which enhances oxidative stress and lipogenesis in NAFLD. The forkhead transcription factor O1 (FoxO1) is another important gene involved in NAFLD, which causes lipid disorders when insulin resistance appears in the liver. However, the interactions between FTO and FoxO1 during the pathogenesis of NAFLD have not been fully elucidated. This study was designed to identify the relationship between these two factors that are involved in the development of NAFLD.

METHODSThis study includes two parts referred to as animal and cell experiments. Twelve female SPF C57BL/6 mice were fed a high-fat diet to serve as an NAFLD animal model. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), total triglyceride (TG), total cholesterol (TC), alkaline phosphatase (ALP), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) were measured. Immunohistochemical analysis was used to detect the expression and histological localization of FTO, FoxO1, and adenosine monophosphate (AMP)-activated protein kinase (AMPK). The L02 cells were exposed to high fat for 24, 48, or 72 hours. Oil red O staining was used to detect intracellular lipid droplets. Reverse transcription-polymerase chain reaction was used for analyzing the levels of FTO and FoxO1 mRNA.

RESULTSAt the end of 10 weeks, ALP, ALT, AST, and LDL were significantly increased (P < 0.01), while TC and TG were also significantly higher (P < 0.05). In addition, HDL was significantly decreased (P < 0.05). The FTO and FoxO1 proteins were weakly expressed in the control group, but both FTO and FoxO1 were expressed significantly higher (P < 0.01) in the experimental group, and the expression of the two factors was significantly correlated. AMPK in the high-fat group showed a low level of correlation with FTO, but not with FoxO1. Oil Red O staining results showed that the cells cultured in 50% fetal bovine serum for 24, 48, or 72 hours exhibited steatosis. FTO and FoxO1 mRNA were increased in the high-fat group compared with the normal group (P < 0.01). The expression levels of FTO and FoxO1 mRNA were the highest at 48 hours (P < 0.05).

CONCLUSIONSA high-fat diet leads to higher expression of FTO, phosphorylation of FoxO1, and decreased phosphorylation of AMPK. These results suggest that the interactions between FTO and FoxO1 are closely related to the pathogenesis of NAFLD.

Alanine Transaminase ; genetics ; metabolism ; Animals ; Aspartate Aminotransferases ; genetics ; metabolism ; Cholesterol ; metabolism ; Diet, High-Fat ; adverse effects ; Disease Models, Animal ; Female ; Forkhead Transcription Factors ; genetics ; metabolism ; Lipoproteins, HDL ; metabolism ; Lipoproteins, LDL ; metabolism ; Liver ; metabolism ; Mice ; Mice, Inbred C57BL ; Non-alcoholic Fatty Liver Disease ; metabolism ; pathology ; Obesity ; metabolism ; pathology ; Triglycerides ; metabolism

OBJECTIVETo observe the effect and safety of Xiaozhi particles, integrated taohong Siwu tang and Erchen tang and Xuezhikang capsule in treating hyperlipidaemia (HLP) associated with highly active antiretroviral therapy (HAART).

METHODIn the multi-centered, randomized controlled clinical study, 180 hyperlipidaemia associated with highly active antiretroviral therapy cases were divided into the treatment group treated by Xiaozhi particles, integrated Taohong Siwu tang and Erchen tang, and the control group treated by Xuezhikang capsule. The treatment course was 12 weeks. The total cholesterol (Tch), triglyceride (TG), low density lipoprotein (LDL) and high-density lipoprotein(HDL) were observed.

RESULTAfter 12 weeks, compared with Xuezhikang capsule, the change difference of Tch, LDL, HDL in the Chinese traditional medicine formula groups of patients is significant (P < 0.05), the change of the TG has no significant difference. The effect of Tch, LDL in Xuezhikang capsule groups is better than in traditional Chinese medicine formula group,but the effect of HDL in traditional Chinese medicine formula group is better than in Xuezhikang capsule groups.

CONCLUSIONIntegrated Taohong Siwu tang and Erchen tang, Xiaozhi particles and Xuezhikang capsule can be used to control the hyperlipidaemia associated with highly active antiretroviral therapy as one of the main Chinese native medicine preparation.

Adult ; Antiretroviral Therapy, Highly Active ; adverse effects ; Cholesterol ; blood ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Humans ; Hyperlipidemias ; blood ; chemically induced ; drug therapy ; Lipoproteins, HDL ; blood ; Lipoproteins, LDL ; blood ; Male ; Triglycerides ; blood

OBJECTIVETo study the effects of high-fat plus ethanol diet on myocardial ultrastructure in rats.

METHODS40 male SD rats in seventy-eight-week old were randomly divided into four groups: group A was control group, fed with common feedstuff; group B was high-fat diet group, freely foraging high-fat feedstuff; group C was ethanol group, the rats were intragastrically administered 60% ethanol solution twice a day by 1 ml/kg; group D was high-fat diet and ethanol group, the rats freely foraged high-fat feedstuff, and ethanol solution was intragastrically administered as before. After 12 weeks, blood samples were taken through jugular vein, the concentration of blood cholesterol (TG), triglycerides (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), apolipoprotein A1 (Apo-A1), apolipoprotein B (Apo-B), and alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL) were determined. The cardiac index was also determined for all groups and the cardiac morphous were observed by high resolution Doppler ultrasound, and myocardial ultrastructure was observed by transmission electron microscope.

RESULTSAfter experiment, TG levels of groups A, B, C, D were (1.07 +/- 0.21), (2.34 +/- 0.72), (1.33 +/- 0.42) and (1.75 +/- 0.65) mmol/L, respectively (F = 8.323, P = 0.000); TC levels were (1.74 +/- 0.38), (5.66 +/- 1.74), (1.70 +/- 0.44) and (5.65 +/- 2.95) mmol/L, respectively (F = 13.670, P = 0.000); HDL levels were (0.65 +/- 0.11), (2.99 +/- 0.54), (0.52 +/- 0.13) and (2.06 +/- 0.26) mmol/L, respectively (F = 112.225, P = 0.000); LDL levels were (0.74 +/- 0.22), (1.87 +/- 0.90), (0.60 +/- 0.26) and (1.54 +/- 0.78) mmol/L, respectively (F = 7.318, P = 0.001); Apo-A1 levels were (0.25 +/- 0.10), (0.31 +/- 0.14), (0.21 +/- 0.05) and (0.36 +/- 0.11) g/L, respectively (F = 3.015, P = 0.047); Apo-B levels were (0.18 +/- 0.03), (0.11 +/- 0.04), (0.16 +/- 0.03) and (0.39 +/- 0.13) g/L, respectively (F = 15.621, P = 0.000); ALT levels were (111.25 +/- 20.18), (447.13 +/- 89.25), (173.13 +/- 44.01) and (198.25 +/- 39.81) U/L, respectively (F = 58.708, P = 0.000); AST levels were (105.50 +/- 9.99), (483.00 +/- 16.80), (120.75 +/- 5.09) and (276.88 +/- 10.48) U/L, respectively (F = 1906.624, P = 0.000);TBIL levels were (1.35 +/- 0.12), (1.66 +/- 0.18), (1.89 +/- 0.15) and (2.68 +/- 0.35)U/L, respectively (F = 55.006, P = 0.000); cardiac indexes were (3.02 +/- 0.22)%, (3.21 +/- 0.16)%, (3.26 +/- 0.26)% and (3.43 +/- 0.27)%, respectively (F = 16.150, P = 0.000). There were changes of cardiac morphous in group C and D, but not in group A and B; the myocardial ultrastructure was normal in Group A, but light to heavy changes were found in group B, C and D.

CONCLUSIONHigh-fat diet and excessive intake of ethanol significantly induce abnormal lipid metabolism. High-fat diet induces the changes of myocardial ultrastructure before cardiac morphous and electrocardiogram, and intake of ethanol changes cardiac muscle in microstructure and macroscopy. High-fat diet plus ethanol may worsen this injury farther.

Animal Feed ; Animals ; Apolipoproteins B ; blood ; Cholesterol, LDL ; blood ; Dietary Fats ; Ethanol ; adverse effects ; Hyperlipidemias ; blood ; pathology ; Lipids ; blood ; Lipoproteins, LDL ; blood ; Male ; Myocardium ; pathology ; ultrastructure ; Rats ; Rats, Sprague-Dawley