Regulation and Role of Adiponectin Secretion in Rat Ovarian Granulosa Cells.

Adiponectin is an important adipokine involved in glucose and lipid metabolism, but its secretion and potential role in regulating glucose utilization during ovarian development remains unclear. This study aims to investigate the mechanism and effects of follicle-stimulating hormones (FSHs) on adiponectin secretion and its following impact on glucose transport in the granulosa cells of rat ovaries. A range of experimental techniques were utilized to test our research, including immunoblotting, immunohistochemistry, immunofluorescence, ELISA, histological staining, real-time quantitative PCR, and transcriptome analysis. The immunohistochemistry results indicated that adiponectin was primarily located in the granulosa cells of rat ovaries. In primary granulosa cells cultured in vitro, both Western blot and immunofluorescence assays demonstrated that FSH significantly induced adiponectin secretion within 2 h of incubation, primarily via the PKA signaling pathway rather than the PI3K/AKT pathway. Concurrently, the addition of the AdipoR1/AdipoR2 dual agonist AdipoRon to the culture medium significantly stimulated the protein expression of GLUT1 in rat granulosa cells, resulting in enhanced glucose absorption. Consistent with these in vitro findings, rats injected with eCG (which shares structural and functional similarities with FSH) exhibited significantly increased adiponectin levels in both the ovaries and blood. Moreover, there was a notable elevation in mRNA and protein levels of AdipoRs and GLUTs following eCG administration. Transcriptomic analysis further revealed a positive correlation between the expression of the intraovarian adiponectin system and glucose transporter. The present study represents a novel investigation, demonstrating that FSH stimulates adiponectin secretion in ovarian granulosa cells through the PKA signaling pathway. This mechanism potentially influences glucose transport (GLUT1) and utilization within the ovaries.

Decoding Noncoding RNAs: Role of MicroRNAs and Long Noncoding RNAs in Ocular Neovascularization.

Ocular neovascularization is a pathological sequel of multiple eye diseases. Based on the anatomical site into which the abnormal neovessels grow, ocular neovascularization can be categorized into corneal neovascularization, choroidal neovascularization, and retinal neovascularization. Each category is intractable, and may lead to blindness if not appropriately treated. However, the current therapeutic modalities, including laser photocoagulation, vitrectomy surgery, and anti-VEGF drugs, raise concerns due to limited efficacy, damage on retinal parenchyma and vasculature, and the patients' unresponsiveness to the treatments. Therefore, the in-depth study on pathogenesis of and the search for novel therapeutic targets to the ocular neovascularization are needed. During the last 10 years or so, a large number of literatures have emerged indicating a critical role of noncoding RNAs, particularly microRNAs (miRNAs) and long noncoding RNAs (lncRNAs), in the pathogenesis and regulation of the ocular neovascularization. This review summarizes the current understanding of the biosynthesis and functions of the miRNAs and lncRNAs, the regulation of the miRNAs and lncRNAs in neovascular eye diseases, as well as the roles of these noncoding RNAs in the disease models of ocular neovascularization, in the hope that it could provide clues for the pathogenesis of and molecular targets to the ocular neovascularization.

[Simultaneous determination of trillin and desgalactotigonin contents in Solanum lyratum by HPLC-ELSD].

OBJECTIVE: To establish a HPLC-ELSD method for simultaneous determination of trillin and desgalactotigonin contents in Solanum lyratum. METHOD: A Diamonsil C18 column (4.6 mm x 250 mm, 5 microm) was adopted, with the mobile phase consisting of acetonitrile-10 mmol x L(-1) ammonium acetate (52: 48). The temperature was 25 degrees C, the flow rate was set at 0.6 mL x min(-1), and the sample size is 20 microL. The temperature of drift tubes and gas flow rate of the detector were set at 95 degrees C and 2.3 L x min(-1), respectively. RESULT: With in the linear ranges of 20-200 mg x L(-1) and 10-100 mg x L(-1), trillin and desgalactotigonin show a good linear relationship. The average recovery was 99.4% (RSD 0.90%) for trillin and 100.3% (RSD 1.1%) for desgalactotigonin. CONCLUSION: The method is so accurate and easily reproducible that it is suitable for the quality control of S. lyratum medicinal materials.

Simultaneous determination of four alkaloids in Solanum lyratum Thunb by UPLC-MS/MS method.

Four alkaloids, strychnine, soladulcidine, solamargine and (3-O-beta-D-glucopyranosyl-(1 --> 2)beta-D-glucopyranosyl-(1 --> 4)-beta-D-galactopyranoside-(25xi)-solanidan-3beta,23beta-diol)(abbreviation, glycoalkaloid A) were isolated from Solanum lyratum Thunb. The structures were elucidated by NMR and measuring physicochemical properties. Then a novel and rapid method using an ultra-performance liquid chromatography coupled with mass spectrometry was developed and validated for the simultaneous determination of these compounds. An acquit UPLC BEH C18 column (2.1 mm x 50 mm, 1.7 microm) was used. Acetonitrile and 0.1% formic acid were adopted as mobile phase. Detection was performed on a Waters Micromass Quattro Premier tandem quadrupole mass spectrometer in the positive ion mode using an electrospray source. The multiple reaction monitoring (MRM) mode was used to detect the target compounds. The established method showed a good linearity (R2 > 0.999 0) over the investigated concentration ranges, good inter-day and intra-day precisions (less than 2%) and good recoveries (from 99.8% to 100.1%) for all four target compounds. Compared to previous methods employing conventional high performance liquid chromatography (HPLC) separation, the ultra-high-pressure liquid chromatography-tandem mass spectrometry achieved preferable chromatographic parameters and significantly increased sample throughput.

Simultaneous determination of five nucleosides and nucleobases of Rehmannia glutinosa Libosch. by high performance liquid chromatography.

This study is to establish a method for simultaneously determination of five nucleosides and nucleobases, including hypoxanthine, uridine, adenine, guanosine and adenosine in Rehmannia glutinosa Libosch. which was collected from different regions in China. A Diamonsil C18 column (250 mm x 4.6 mm, 5 microm) was used. Acetonitrile and 0.04 mol L(-1) potassium dihydrogen phosphate solution were adopted as mobile phase with gradient elution. The flow rate was 1 mL min(-1) and column temperature was 30 degrees C. The detection wavelength was at 254 nm. The method had good linearity over the range of 1.0 - 16.0 microg mL(-1) (r2 = 0.999 8), 5.0 - 80.0 microg mL(-1) (r2 = 0.999 8), 1.0 - 16.0 microg mL(-1) (r2 = 0.999 5), 1.25 - 20.0 microg mL(-1) (r2 = 0.999 8) and 1.0 - 16.0 microg mL(-1) (r2 = 0.999 8) for hypoxanthine, uridine, adenine, guanosine and adenosine, respectively. The average recoveries were between 98.8% and 100.7%. The content of hypoxanthine, uridine, adenine, guanosine and adenosine in Rehmannia glutinosa Libosch. from different regions was significantly different. This established method was sensitive and reliable for the quantification of five chemical constituents in Rehmannia glutinosa Libosch.

[Pharmacokinetics and relative bioavailability of KC-404 sustained release tablets in healthy volunteers].

OBJECTIVE: To study the pharmacokinetics and relative bioavailability of KC-404 sustained release tablets and capsules in healthy volunteers. METHODS: The concentration of KC-404 in serum was determined by HPLC method after a single oral dose (20 mg) of tablet or capsule was administered to 19 healthy male volunteers respectively in an open randomized cross-over test. RESULTS: After being processed by 3P87 pharmacokinetics program, the experiment data showed that the pharmacokinetic parameters of the tablets and the capsules were AUC0-->infinity: 740.12 ng.h/ml and 724.04 ng.h/ml; tmax: 5.37 h and 5.11 h; Cmax: 46.98 ng/ml and 46.29 ng/ml; T1/2: 7.4 h and 7.0 h; MRT0-->infinity: 14.78 h and 14.39 h respectively. There were no significant differences in AUC, tmax, Cmax, T1/2 and MRT0-->infinity between these two preparations (P > 0.05). The relative bioavailability of KC-404 sustained release tablets was 102.6%. CONCLUSION: These two preparations are bioequivalent.