Study on anti-hyperlipidemia effect of Linderae Radix via regulating reverse cholesterol transport / 中国中药杂志

This article aims to investigate the ameliorative effect of Linderae Radix ethanol extract on hyperlipidemia rats induced by high-fat diet and to explore its possible mechanism from the perspective of reverse cholesterol transport(RCT). SD rats were divided into normal group, model group, atorvastatin group, Linderae Radix ethanol extract(LREE) of high, medium, low dose groups. Except for the normal group, the other groups were fed with a high-fat diet to establish hyperlipidemia rat models; the normal group and the model group were given pure water, while each administration group was given corresponding drugs by gavage once a day for five weeks. Serum total cholesterol(TC), triglyceride(TG), high density lipoprotein-cholesterol(HDL-c), low density lipoprotein-cholesterol(LDL-c), alanine aminotransferase(ALT), and aspartate aminotransferase(AST) levels were measured by automatic blood biochemistry analyzer; the contents of TC, TG, total bile acid(TBA) in liver and TC and TBA in feces of rats were detected by enzyme colorimetry. HE staining was used to observe the liver tissue lesions; immunohistochemistry was used to detect the expression of ATP-binding cassette G8(ABCG8) in small intestine; Western blot and immunohistochemistry were used to detect the expression of peroxisome proliferator-activated receptor gamma/aerfa(PPARγ/α), liver X receptor-α(LXRα), ATP-binding cassette A1(ABCA1) pathway protein and scavenger receptor class B type Ⅰ(SR-BⅠ) in liver. The results showed that LREE could effectively reduce serum and liver TC, TG levels, serum LDL-c levels and AST activity, and increase HDL-c levels, but did not significant improve ALT activity and liver index; HE staining results showed that LREE could reduce liver lipid deposition and inflammatory cell infiltration. In addition, LREE also increased the contents of fecal TC and TBA, and up-regulated the protein expressions of ABCG8 in small intestine and PPARγ/α, SR-BⅠ, LXRα, and ABCA1 in liver. LREE served as a positive role on hyperlipidemia model rats induced by high-fat diet, which might be related to the regulation of RCT, the promotion of the conversion of cholesterol to the liver and bile acids, and the intestinal excretion of cholesterol and bile acids. RCT regulation might be a potential mechanism of LREE against hyperlipidemia.

circ_0000285 regulates VAMP2 through sponging miR-127 and participates in the development of ovarian cancer / 中华内分泌外科杂志

Objective:To investigate the effects of circ_0000285/miR-127/VAMP2 axis on proliferation and invasion of ovarian cancer (OC) cells.Methods:Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of circ_0000285, miR-127 and VAMP2 in tissue with OC and normal tissue respectively, and the correlation circ_0000285/miR-127/VAMP2 was analyzed by correlation analysis. Dual luciferase reporter assay was used to verify the targeting relationship between miR-127 and circ_0000285/VAMP2. Cell proliferation and invasion were subsequently detected by MTT and Transwell assay.Results:Compared with the paracancerous tissue, the expression of miR-127 was significantly decreased, while the expression of circ_0000285 and VAMP2 was increased in OC tissue. miR-127 was negatively correlated with circ_0000285 and VAMP2 ( r=-0.534 8, P<0.000 1; r=-0.376 6, P<0.000 1) . Compared with proliferative activity at 24 h, 48 h, 72 h (0.47±0.03) (0.79±0.05) (1.16±0.09) and invasion (100.00±12.33) in si-NC group, knocking down of circ_0000285 inhibited the proliferation (0.28±0.02) (0.51±0.04) (0.78±0.06) and invasion (49.22±5.08) of OC cells (all P<0.05) . Overexpression of miR-127 could also restrain proliferation, invasion of OC cells, but this effect can be partly saved by circ_0000285. The regulation circ_0000285 on OC may be realized through miR-127/VAMP2. Conclusions:circ_0000285 participates in the progression of ovarian cancer via regulating miR-127/VAMP2 axis. Down-regulating the expression of circ_0000285 inhibits the proliferation and invasion of ovarian cancer cells, circ_0000285 is expected to play a role in the targeted therapies of OC.

Correlation between lipid-lowering efficacy and components of Pericarpium Citri Reticulatae / 中国中药杂志

Based on pharmacodynamics-component correlation analysis,the best effective part of hyperlipidemia of Pericarpium Citri Reticulatae( PCR) was screened out to confirm the possible constituents with the lipid regulating effect,in order to provide a basis for the development of new drugs. Hyperlipidemia rats induced by fat emulsion were used to screen out the best hypolipidemic parts of PCR with TC,LDL-c as the index. HPLC-ESI-MS were used to analyze common constituents of the different solvent extracts from PCR. The constituents were classified and identified based on the retention time,m/z and UV spectra. And the HPLC-DAD were used to determine the contents of flavonoids( narirutin,hesperidin,didymin,nobiletin,tangeretin,3,5,6,7,8,3',4'-heptemthoxyflavone).Correlation analysis was conducted on the constituents and efficacy with the method of SPSS ANOVA bivariate correlation. Five extracts could significantly decrease the content of TC,LDL-c of hyperlipemia rats induced by fat emulsion,and the best effective part were95% ethanol and ethyl acetate. There were 19 common peaks in five different solvent extracts from PCR,and 17 flavonoids were identified and classified,including 10 polymethoxyflavonoids and 7 other flavonoids. According to the raw material quantity,the order of content of flavonones arranged from high to low: n-butyl alcohol part> 95% ethanol part> water part> ethyl acetate part> petroleum ether part; and the order of PMFs arranged from high to low: n-butyl alcohol part≈95% ethanol part≈ethyl acetate part > petroleum ether part > water part. The decreased percentage of TC,LDL-c was positively correlated with 10 common PMFs constituents,which suggested that PMFs may be the effective components for reducing blood lipid.

Anti-lung cancer mechanisms of diterpenoid tanshinone via endoplasmic reticulum stress-mediated apoptosis signal pathway / 中国中药杂志

At present, lung cancer ranks second and first respectively in the incidence and the mortality among malignant tumors. It is urgent to find new effective anti-lung cancer drugs with less side effects and relatively defined mechanisms. Endoplasmic reticulum stress (ERS)-mediated apoptosis pathway is an effective way to promote tumor cell apoptosis; diterpenoid tanshinone (DT), an effective part separated from Salviae Miltiorrhizae Radix et Rhizoma, was found to have an anti-lung cancer effect in previous studies via ERS-induced PERK-EIF2α pathway. In this paper, human lung adenocarcinoma PC9 cell line and nude mouse transplantation tumor model were applied to verify the anti-lung cancer effect of DT in vivo and in vitro, and illuminate the potential mechanism via ERS induced IRE1α/caspase 12 apoptosis pathway. The results showed that in vivo, DT could promote PC9 cell apoptosis in a concentration-dependent manner, up-regulate Bip, IRE1 and TRAF2 protein expressions in tumor tissue, reduce tumor weight and alleviate bodyweight loss. In vitro, DT inhibited the proliferation of PC9 cell line in a concentration-dependent manner, and destroyed the structure of mitochondria in PC9 cell, promoted Bax, IRE1α, Bip, TRAF2 and caspase 12 protein expressions, lower Bcl-2 protein expression in a time-dependent manner. DT shows a good effect on anti-lung cancer both in vivo and in vitro. The mechanism is related to the activation of ERS-induced IRE1α/caspase 12 apoptosis pathway and the promotion of cell apoptosis. ERS-mediated apoptosis pathway may be an important target of DT on anti-lung cancer.

Effects of lotus leaf on inflammatory factors and liver AdipoR2 expressions in rats with NAFLD induced by high fat diet and high glucose / 中国中药杂志

Lotus leaf (LL) is one of the traditional Chinese herbs which can be used for both pharmaceutical and food application, and it posses lipid regulating efficacy. To observe the effect of LL on experimental nonalcoholic fatty liver disease (NAFLD) and its potential mechanism, a NAFLD model was established by feeding SD rat with high-fat and high-glucose diet. LL was administrated to rats in experiment group at the same time. AST,ALT,Cr,BUN,GLU levels in serum were determined by automatic biochemical analyser and TNF-α,IL-6,INS,ADPN,LEP and liver NF-κB,TGF-β1 levels were determined by ELISA according to the specification of the kits. HE staining was applied for histopathological examination and RT-PCR,Western blot was applied for AdipoR2 mRNA and protein expression．Results have shown that LL could significantly decrease ALT,AST,IL-6 level in serum and NF-κB,TGF-β1 level in liver,promote adiponectin content in serum and AdipoR2 protein expression in liver and could alleviate hepatocyte lipid degeneration. These results indicating that LL has protective effect for NAFLD induced by high-fat and high-glucose diet via promoting AdipoR2 expression, improving insulin resistance and inhibiting inflammatory reaction.

Studies on HSCCC preparation technology of diterpenoidtanshinone and its antitumor activity in vitro / 中草药

To optimize the solvent system of high-speed counter-current chromatography (HSCCC) for diterpenoidtanshinone separation and define it's antitumor activity in vitro. Total diterpenoidtanshinone was made by CO2 supercritical fluid extraction (SFE), UPLC was applied to determining the peak area of tanshinone IIA and cryptotanshinone in different solvents up or under phase, and the partition coefficient K values of them were calculated. CKK-8 was used to observe the inhibitory effects of diterpenoidtanshinone on human liver cancer (QGY-7703), lung cancer (PC9, A549), gastric cancer (MKN-45, HGC-27), colon cancer (HCT116), myeloma (U266, RPMI8226), and breast cancer (MCF-7). The best solvent system for HSCCC was petroleum ether-ethyl acetate-methanol-aqua (12∶8∶ 13∶7). The yield of diterpenoidtanshinone was 8.65%. Diterpenoidtanshinone has good effect of antitumor in vitro especially on human PC9 cell. The selected solvent system is suitable for diterpenoidtanshinone separation by HSCCC, and the established HSCCC method is reliable and easy for operating. Diterpenoidtanshinone has good antitumor effect in vitro.

Relationship between ouabain and asthenozoospermia / 华中科技大学学报（医学）（英德文版）

A growing number of researches have shown that ouabain can regulate mammalian sperm function and male reproduction by modulating the sperm motility, capacitation and acrosome reaction in vitro. This study further examined the relationship between ouabain and asthenozoospermia. In this study, the rat was intraperitoneally injected with ouabain at different concentrations (low-dose ouabain group: 12.5 μg/kg body weight per day, and high-dose ouabain group: 25 μg/kg body weight per day) for 30 days to establish the asthenozoospermia model. The sperms from 60 males with normal fertility were incubated with ouabain of gradient concentrations (10(-7)-10(-2) mol/L) for 4 h. The sperm motility was evaluated under a microscope. Moreover, the endogenous ouabain (EO) level was determined in seminal plasma of mild or severe asthenozoospermia patients and males with normal fertility by competitive inhibition ELISA. The results showed that the sperm motility was significantly diminished in the rats treated with different concentrations of ouabain. The number of motile sperms (grades a and b) was decreased greatly in a time- and dose-dependent manner in 10(-5)-10(-2) mol/L ouabain groups (P0.05). Furthermore, the EO level was significantly increased in asthenozoospermia patients as compared with that in males with normal fertility (25.27±1.71 μg/L in mild asthenozoospermia patients, 26.52±1.82 μg/L in severe asthenozoospermia patients, 19.31±1.45 μg/L in normal fertility men) (P<0.01). In conclusion, rat asthenozoospermia was successfully established by intraperitoneal injection of ouabain, and 10(-5) mol/L ouabain was sufficient enough to inhibit sperm motility in vitro. Moreover, EO, a normal constituent of seminal plasma, was highly expressed in asthenozoospermia males as compared with normal fertility ones.

Relationship between ouabain and asthenozoospermia / 华中科技大学学报（医学）（英德文版）

A growing number of researches have shown that ouabain can regulate mammalian sperm function and male reproduction by modulating the sperm motility, capacitation and acrosome reaction in vitro. This study further examined the relationship between ouabain and asthenozoospermia. In this study, the rat was intraperitoneally injected with ouabain at different concentrations (low-dose ouabain group: 12.5 μg/kg body weight per day, and high-dose ouabain group: 25 μg/kg body weight per day) for 30 days to establish the asthenozoospermia model. The sperms from 60 males with normal fertility were incubated with ouabain of gradient concentrations (10(-7)-10(-2) mol/L) for 4 h. The sperm motility was evaluated under a microscope. Moreover, the endogenous ouabain (EO) level was determined in seminal plasma of mild or severe asthenozoospermia patients and males with normal fertility by competitive inhibition ELISA. The results showed that the sperm motility was significantly diminished in the rats treated with different concentrations of ouabain. The number of motile sperms (grades a and b) was decreased greatly in a time- and dose-dependent manner in 10(-5)-10(-2) mol/L ouabain groups (P<0.01), while no obvious change in sperm motility was observed in 10(-7)-10(-6)mol/L groups even for 4-h incubation (P>0.05). Furthermore, the EO level was significantly increased in asthenozoospermia patients as compared with that in males with normal fertility (25.27±1.71 μg/L in mild asthenozoospermia patients, 26.52±1.82 μg/L in severe asthenozoospermia patients, 19.31±1.45 μg/L in normal fertility men) (P<0.01). In conclusion, rat asthenozoospermia was successfully established by intraperitoneal injection of ouabain, and 10(-5) mol/L ouabain was sufficient enough to inhibit sperm motility in vitro. Moreover, EO, a normal constituent of seminal plasma, was highly expressed in asthenozoospermia males as compared with normal fertility ones.

Simultaneous determination of four glycosylflavones from Lophatherum gracile by RP-HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To develop a RP-HPLC method for simultaneous determination of orientin, isorientin, vitexin and isovitexin in Lophatherum gracile from different habitat and harvesting time.</p><p><b>METHOD</b>The HPLC method was applied and the chromatographic column was a Waters XBridge C18 column (4.6 mm x 250 mm, 5 microm). The mobile phase consisted of methanol-0.05% acetic acid (35:65). The flow rate was 1.0 mL min(-1) and the detection wavelength was set at 340 nm. The column temperature was set at 25 degrees C.</p><p><b>RESULT</b>Four components were isolated well, the linear relationships were excellent. The mean recoveries and RSD values of orientin, isorientin, vitexin and isovitexin were 103.2%, 2.1%; 101.6%, 2.7%; 98.4%, 2.3%; 99.2%, 1.8%, respectively.</p><p><b>CONCLUSION</b>The HPLC method is simple, sensitive and reliable, and can be used for the quality control of the medicinal material.</p>