Silencing of Myh3 Gene by siRNA Inhibits Glycolysis in C2C12 Cells / 中国生物化学与分子生物学报

The Myh3 (myosin heavy chain 3) gene is a marker gene of muscle cell differentiation and regulates the utilization of energy in muscle cells, but whether it affects the glycolysis process of muscle cells in different states is rarely reported. In this paper, the expression patterns of Myh3 and glycolysis-related genes Pkm (M-type pyruvate kinse), Prkag3 (protein kinase adenosine monophosphate-activated γ3-subunit), and Gsk3β (glycogen synthase kinase-3β) were studied by the qRT-PCR (quantitative-Real-Time-PCR) method using C2C12 cells at different stages of myoblast and adipogenic differentiation as models. It was found that in the process of myoblast differentiation of C2C12 cells, the relative expression trends of Myh3 and glycolysis genes Prkag3 and Pkm were basically the same, and the relative expression levels first increased, reached the peak on the second day of differentiation, and then decreased; glycogen synthase the expression trend of the inhibitory gene Gsk3β was relatively stable. In the process of adipogenic differentiation of C2C12 cells, the relative expression trend of Myh3 and glycolysis genes Prkag3 and Pkm remained basically the same, and the relative expression gradually increased, reaching the highest value on the 8th day of differentiation; glycogen synthase inhibitory gene Gsk3β expression remained stable. In the myogenic differentiation state of C2C12 cells, qRT-PCR and Western blotting were used to detect the effects of interfering Myh3 on the mRNA and protein expressions of glycolysis-related genes Pkm, Prkag3, and Gsk3β. The results showed that after interfering with Myh3, the mRNA expressions of glycolysis genes Pkm and Prkag3 were significantly decreased (P 0.05). The protein levels of Myh3 and Pkm were significantly lower than those in the blank group and NC group. Under the adipogenic differentiation state of C2C12 cells, after interfering with Myh3, the mRNA levels of glycogen synthase inhibitor Gsk3β and glycolysis gene Prkag3 were significantly increased (P<0.01), and the mRNA level of glycolysis gene Pkm was decreased; the protein levels of Myh3 and Pkm in the Myh3 interference group were also lower than those in the blank group and NC group. Based on the above studies, there are significant differences in the levels of glycolysis in C2C12 cells in the myogenic and adipogenic states, and the expression patterns of Myh3 and glycolysis genes are similar. Further results showed that Myh3 suppression could inhibit the glycolysis of C2C12 cells in the myogenic state without affecting the glycogen synthesis. Unlike in the myogenic state, interfering expression of Myh3 in the adipogenic state of C2C12 cells inhibited both glycogen synthesis and glycolysis.

Metabonomics research strategy based on microdialysis technique / 中国中药杂志

Metabonomics is the branch of systems biology. It has been widely used in the fields of diagnostic markers discovery, disease prognosis, drug action mechanism, drug efficacy and toxicity evaluation, traditional Chinese medicine syndromes differentiation. There are shortcomings in the conventional metabonomics research. Microdialysis technology is a new type of biosampling technology, and metabonomics research based on microdialysis technology is in the ascendant. In view of the particularity of microdialysis technology and its great differences from traditional sampling and pretreatment methods, the metabonomics process based on microdialysis technology has certain similarities with traditional metabonomics research, and its basic process has some particularity. Advantages and basic strategies of metabonomics research by microdialysis technology are systematically summarized for researchers' reference.

Plasma Lipidomics of Coronary Heart Disease Patient with Syndrome of Intermingling of Phlegm and Static Blood Based on UPLC-Q/TOF-MS / 中国实验方剂学杂志

<b>Objective::To study on the plasma lipidomics characteristics of coronary heart disease (CHD) patients with syndrome of of intermingling of phlegm and static blood, and to find differential lipid metabolites between them and healthy volunteers. <b>Method::The plasma samples from CHD patients with syndrome of intermingling of phlegm and static blood and healthy volunteers of the same age were collected. The plasma lipidomics was carried out by UPLC-Q/TOF-MS. The plasma samples were detected under positive and negative ion modes, and the primary and secondary mass spectrometry datas were collected simultaneously, and the <italic>m</italic>/<italic>z</italic> ranges were 100-2 000 and 50-2 000, respectively. The lipidomics model was established by orthogonal partial least squares discriminant analysis (OPLS-DA). Differential lipid metabolites were identified based on multivariate statistics. <b>Result::OPLS-DA model could obviously distinguish CHD patients with syndrome of intermingling of phlegm and static blood and healthy volunteers. A total of 15 plasma differential lipid metabolites were identified, such as C16 sphinganine, phytosphingosine, <italic>N</italic>, <italic>N</italic>-dimethyl-safingol, 2-hydroxyphytanic acid, orotinichalcone, PC[18∶2(2<italic>E</italic>, 4<italic>E</italic>)/0∶0], PC(0∶0/16∶0), epitestosterone sulfate, etiocholanolone sulfate, PS[22∶1(11<italic>Z</italic>)/0∶0], PC[16∶0/20∶4(5<italic>E</italic>, 8<italic>E</italic>, 11<italic>E</italic>, 14<italic>E</italic>)], PC[19∶1(9<italic>Z</italic>)/17∶2(9<italic>Z</italic>, 12<italic>Z</italic>)], PC(16∶0/0∶0), PC(18∶0/0∶0), PS[15∶1(9<italic>Z</italic>)/22∶1(11<italic>Z</italic>)]. <b>Conclusion::There are significant differences in plasma lipid characteristics between CHD patients with syndrome of intermingling of phlegm and static blood and healthy volunteers. The plasma differential lipid metabolites are helpful for the accurate differentiation of CHD patients with syndrome of intermingling of phlegm and static blood.

Brain lipidomics of intervention effect of Salviae Miltiorrhizae Radix et Rhizoma and Chuanxiong Rhizoma on ischemic stroke based on UPLC-Q-TOF-MS technique / 中国中药杂志

The aim of this paper was to investigate the intervention effect of Salviae Miltiorrhizae Radix et Rhizoma and Chuanxiong Rhizoma on brain lipid metabolism in rats with ischemic stroke. The ischemic stroke rat model was established by middle cerebral artery occlusion( MCAO) method. The brain tissues were collected after the last administration with Salviae Miltiorrhizae Radix et Rhizoma and Chuanxiong Rhizoma decoction lyophilizate. UPLC-Q-TOF-MS was used to carry out the brain lipidomics study. The lipidomics data were processed with the OPLS-DA model to find out the lipid regulation effect of Salviae Miltiorrhizae Radix et Rhizoma combined with Chuanxiong Rhizoma on ischemic stroke. The results showed that Salviae Miltiorrhizae Radix et Rhizoma and Chuanxiong Rhizoma decoction lyophilized powder can significantly alleviate brain lipidomics profiles in middle cerebral artery occlusion model rats. Eleven differential lipid metabolites in ischemic stroke model were identified. In this experiment,the protective effects of Salviae Miltiorrhizae Radix et Rhizoma and Chuanxiong Rhizoma decoction lyophilized powder on cerebral ischemia injury was verified,which might be related to the regulation of abnormal lipid metabolism.

Study of pharmacokinetics of nicotine in local brain by using microdialysis and stable labeled isotope / 药学学报

The paper is to report the study of pharmacokinetics of transdermal administered nicotine in the brain of freely moving rat by using microdialysis with stable labeled isotope as internal standard. The pharmacokinetic behavior of nicotine in Sprague Dawley rat brain was investigated after intranasal administration (3.75 mg). Brain fluid samples were collected by intracerebral microdialysis with DL-nicotine as internal standard. Concentrations of nicotine and DL-nicotine in the sample were measured by HPLC-MS/MS. Main pharmacokinetic parameters were calculated and analyzed by Das 2.0 pharmacokinetic software. The recovery of nicotine and the delivery of DL-nicotine were the same. The fate of absorption and distribution was two compartment model and the values of t1/2alpha was 170.31 min, t1/2beta was 263.30 min and the AUC(0-infinity) was 2.75 x 10(5) microg x L(-1) min separately. DL-nicotine can be used to calibrate the recovery of nicotine, and the new method of stable isotope microdialysis can be used to study the pharmacokinetics of freely moving rat. It will make sense for the treatment of addiction of tobacco and provide a new thought for the research of pharmacokinetics-pharmacodynamic combination.

Absorption of extractive Rhizoma Coptidis in rat everted gut scas / 中国中药杂志

<p><b>OBJECTIVE</b>Using in vitro everted gut seas to research the intestinal absorption of the extractive Rhizoma Coptidis at the different intestinal section and the different density.</p><p><b>METHOD</b>Berberine (BER) and palmatine (PAL) which are representative compositions of the extractive Rhizoma Coptidis in everted gut seas are detected by HPLC, and calculated the absorption parameter to describe the character of absorption.</p><p><b>RESULT</b>The absorption of BER and PAL is linearity in different intestine and different dose, and the square of coefficient correlation exceed 0.9, which consistent with zero order rate process. The K(a) of BER and PAL increases along with the raised dosage of the extractive Rhizoma Coptidis (P < 0.05), indicated it is the passive absorption. The absorption of BER and PAL in the jejunum is the most quick, the ileum and colon are slower.</p><p><b>CONCLUSION</b>In the different dosage of extractive Rhizoma Coptidis, the absorption of BER and PAL Conforms to the zero order rate process at the different intestine, and is the passive absorption.</p>

Determination in vitro of rat plasma protein binding rate of sinomenine by using microdialysis method / 药学学报

<p><b>AIM</b>To determine in vitro the rat plasma protein binding rate by using microdialysis method.</p><p><b>METHODS</b>The binding rate was determined by using microdialysis probe as sampling tools and zero-net flux method as calibrating method. The regression equation was made by the difference of concentrations between the dialysis sample and the perfusate. The x-intercept of regression equation was the free drug concentration (Cf). The plasma protein binding rate was calculated by using the following equation: f = ( C0 - Cf)/C0.</p><p><b>RESULT</b>The binding rate was kept relatively stable in the studied concentration range.</p><p><b>CONCLUSION</b>It is feasible that the plasma protein binding rate can be determined by using microdialysis method.</p>