Metformin inhibits estradiol and progesterone-induced decidualization of endometrial stromal cells by regulating expression of progesterone receptor, cytokines and matrix metalloproteinases.

BACKGROUND: Polycystic ovary syndrome (PCOS) is a serious threat for reproductive-aged women. Metformin has been used for the treatment of PCOS. However, its molecular mechanism in decidualization process of PCOS has not been well featured. METHODS: RT-qPCR analysis was used to detect expression patterns of progesterone receptor (PGR), estradiol receptor alpha (ERα), Cytokeratin 8 and Vimentin in endometrial tissues of PCOS and non-PCOS patients. RT-qPCR assay was also employed to determine mRNA expression of prolactin, Insulin-like growth factor-binding protein 1 (IGFBP-1), matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP9). Cytokine secretion were measured by matching ELISA kits. Protein expression of p-ERK1/2, ERK1/2, p-p38 MAPK, p38 MAPK, and PGR (PGRA and PGRB) was tested by western blot assay. RESULTS: PGR expression was upregulated in PCOS patients. Metformin alleviated estradiol (E2) and progesterone (P4) (EP)-induced decidualization of endometrial stromal cells. Abnormal cytokine secretion was observed in EP-stimulated endometrial stromal cells in the absence or presence of metfromin. Metformin suppressed EP-induced MMP-2 and MMP-9 upregulation. Metformin alleviated EP-triggered p38 MAPK inactivation and PGR (PGRA and PGRB) expression. Metfromin had no effect on ERK1/2 signaling in EP-stimulated endometrial stromal cells. CONCLUSION: Metformin alleviated EP-induced decidualization of endometrial stromal cells by modulating secretion of multiple cytokines, inhibiting expression of MMP-2 and MMP-9, activating p38-MAPK signaling and reducing PGR expression, providing a deep insight into the molecular basis of metfromin therapy for PCOS patients.

Effect of high-level spinal cord injury on myocardial energy metabolism in rats / 中华麻醉学杂志

Objective To evaluate the effect of high-level spinal cord injury (SCI) on the myocardial energy metabolism in rats.Methods Sixty healthy male Sprague-Dawley rats,weighing 250-300 g,were randomly divided into 2 groups (n=30 each) using a random number table:sham operation (group S) and SCI group.SCI was induced in anesthetized rats by dropping a 10 g weight onto C7 spinal cord from 5 cm height falling freely inside a vertical hollow glass tube.At 6,12,24,48 and 72 h after SCI,6 rats in each group were chosen and arterial blood samples were taken for measurement of serum creatine kinase (CK) and creatine kinase isoenzyme-MB (CK-MB) activities.The rats were then sacrificed and myocardial specimens were obtained for examination of myocardial ultrastructure and for determination of ATP weight ratio,levels of Na+-K+-ATPase,Ca2+-Mg2+-ATPase,non-esterified fatty acids (NEFA) and lactic acid (LD),and expression of peroxisome proliferator-activated receptor alpha (PPARα) mRNA and protein (using fluorescent quantitative PCR and Western blot).Results Compared with group S,the serum CK and CK-MB activities were significantly increased,the ATP weight ratio,activities of Na+-K+-ATPase and Ca2+-Mg2+-ATPase and levels of NEFA and LD were decreased,and the expression of PPAR-α mRNA and protein was down-regulated in SCI group.No pathological changes of myocardium were found in group S,and the pathological changes of myocardium were obvious in SCI group.Conclusion High-level SCI can lead to decrease in the myocardial energy metabolism in rats,and down-regulated expression of PPARα is involved in the mechanism.

Effect of creatine phosphate sodium on myocardial protection and calcium-sensitive receptor expression after high-level spinal cord injury / 中华创伤杂志

Objective To investigate the effect of creatine phosphate sodium on myocardial protection and calcium-sensitive receptor (CaSR) expression following high-level spinal cord injury.Methods Thirty healthy male SD rats weighing 250-300 g were assigned to sham operation,12-hour injury,24-hour injury,12-hour injury followed by a single intraperitoneal injection of creatine phosphate sodium,and 24-hour injury followed by a single intraperitoneal injection of creatine phosphate sodium according to the random number table,with 6 rats in each group.High-level spinal cord injury was induced at C7 segment by dropping a 10 g weight falling freely along the hollow glass tube from a 5 cm height.Level of blood troponin Ⅰ (cTnⅠ) was measured.Myocardial tissues were collected to study ultrastructure of myocardial cells under transmission electron microscope and CaSR expression using fluorescence quantitative PCR and Western blotting.Results cTnⅠ level was (0.031 ±0.002) U/L and (0.026 ± 0.001) U/L in 12-and 24-hour injury groups,but it was reduced to (0.023 ± 0.002) U/L and (0.018 ± 0.006) U/L at the same time point in treatment groups (P ＜ 0.05).Whereas either in injnry or treatment groups,cTnⅠ level was higher than (0.004 ± 0.002) U/L in sham operation group (P ＜ 0.05).CaSR mRNA level was (0.991 ±0.146) × 10-3 and (1.245 ±0.204) × 10-3 in 12-and 24-hour injury gronp and decreased to (0.880 ± 0.096) × 10-3 and (0.782 ± 0.138) × 10 3 at the same time point in treatment groups (P ＜ 0.05),but all were higher than (0.437 ± 0.065) × 10-3 in sham operation group (P ＜ 0.05).CaSR protein expressed in 12-and 24-hour injury group was (0.627 ±0.066) × 10 3 and (0.809 ±0.154) ×10 3 and lowered to (0.505 ±0.176) × 10-3 and (0.524 ±0.138) × 10-3 at the same time point in treatment groups,but all were higher than (0.331 ± 0.102) × 10-3 in sham operation group (P ＜ 0.05).Transmission electron microscopy demonstrated normal myocardial ultrastructure in sham operation group but impairment in injury groups,but the impairment was significantly improved in treatment groups.Conclusion Creatine phosphate sodium can decrease cTnⅠ level,attenuate the damage to myocardial ultrastructure and down-regulate CaSR after high-level spinal cord injury.

The inhibition effect of high storage temperature on the recrystallization rate during dissolution of nimodipine-Kollidon VA64 solid dispersions (NM-SD) prepared by hot-melt extrusion.

Nimodipine (NM) solid dispersions (SD) were prepared by hot-melt extrusion with polyvinylpyrrolidone/vinyl acetate copolymer (PVP-VA; Kollidon VA64) as the carrier, then the extrudate was stored at different temperatures (20°C, 40°C, 60°C) after milling. Conventional differential scanning calorimetry and powder X-ray diffraction revealed that the drug was present as an amorphous state in the carrier. The dissolution curves of NM from SD at 37°C in water showed that the rate of recrystallization for NM from SD stored at high temperatures was slower than that from room temperature. The single glass transition temperature (T(g) ) measured by modulated temperature DSC remained constant around 89°C and indicated that the amorphous state of the compound was not altered by heating. However, the C=O vibration of the amide function of Kollidon VA64 increased from 1658 to 1675 cm(-1) was observed in infrared spectra and implied the weakness of H-bonding between the compound and the polymer. The above-mentioned experiments clued to us that suitable postcooking for a short time is good for SD; however, the mechanisms of the changes were still not clear and need further investigation.

Nimodipine (NM) tablets with high dissolution containing NM solid dispersions prepared by hot-melt extrusion.

Using a mixture of Eudragit EPO and polyvinylpyrrolidone/vinyl acetate copolymer (PVP/VA) (Kollidon VA64) as carriers, a nimodipine solid dispersion (NM-SD) was prepared by hot-melt extrusion (HME) to achieve high dissolution. The dissolution profiles in 900 mL 0.1 mol/L HCl showed that the drug release of NM-SD reached 90% in 1h. Powder X-ray diffraction (PXRD) and differential scanning calorimetry (DSC) were used to characterize the state of NM. The results obtained showed that NM was in an amorphous form in the solid dispersion (SD). NM-SD tablets (NM-T-SD) were compressed by wet granulation and direct compression, respectively. The stability of NM-T-SD was examined during a 2-month storage period (40 degrees C, RH 75%). The results showed that the dissolution of NM-T-SD was slightly reduced after 2 months storage (40 degrees C, RH 75%), which implied that aging occurred to some degree. However, no NM crystals could be observed by PXRD after 2 months storage for NM-T-SD (F11) prepared by direct compression.