Lin28A alleviates ovariectomy-induced osteoporosis through activation of the AMP-activated protein kinase pathway in rats.

AIM: To investigate the role of Lin28A in ovariectomy-induced osteoporosis and to elucidate the underlying molecular mechanism. METHODS: Bilateral ovariectomy was conducted to generate an ovariectomy (OVX) rat model. Western blotting was performed to assess the relative expression levels of Lin28A, osteocalcin (OCN), runt-related transcription factor 2 (RUNX2), adenosine monophosphate-activated protein kinase (AMPK) and phosphorylated AMPK (p-AMPK) proteins. Enzyme-linked immunosorbent assays were performed to detect the serum levels of calcium, E2, alkaline phosphatase (ALP) and interleukin (IL)-1ß. Three-point bending test was used to assess biomechanical parameters of left femoral diaphysis. Hematoxylin and eosin (HE) staining was conducted to detect the trabecular structure of bone tissue. Dihydroethidium assay kit was used to measure the intracellular reactive oxygen species (ROS) level in osteoclasts. Alizarin red staining revealed the calcium deposit in bone marrow stromal cells (BMSC). RESULTS: The expression levels of Lin28A, OCN, RUNX2, AMPK and p-AMPK proteins were significantly decreased in OVX rats. The serum levels of calcium, E2, ALP and IL-1ß were significantly declined in OVX rats. Biomechanical parameters of left femoral diaphysis were significantly decreased in OVX rats. OVX-induced trabecular abnormalities. ROS level was dramatically increased in the bone tissue of OVX rats, and calcium deposit was dramatically decreased in BMSC cells of OVX rats. These effects induced by OVX could be prevented by overexpression of Lin28A. CONCLUSION: Lin28A alleviates ovariectomy-induced osteoporosis through activation of AMPK pathway in rats.

Bruguiera gymnorrhiza (L.) Lam. Fruit Accelerates Healing in Gastric Injury via the Regulation of the NF-κB Pathway.

Objective: The present study aimed at the anti-inflammatory and antioxidant effects of the extract of Bruguiera gymnorrhiza (L.) Lam. fruit (BGF) on the gastric injury. Materials and Methods: The chemical components in the extract of BGF were used in UPLC/Q-Orbitrap analysis. 60 SD rats were randomized into six groups: normal group (MC), ethanol-injured control group (EC), omeprazole group, and three groups with different doses (50, 100, and 200 mg/kg) of BGF. After continuous administration for seven days, the stomachs of rats were taken out to observe the pathological gastric tissue changes; inflammatory factors and oxidative stress markers in the stomach tissues were measured. Western blot (WB) analyses were conducted to explore the mechanism of BGF on gastric tissue and RAW 246.7 cells with excessive inflammation. Results: BGF enhanced gastric mucosal protection by improving the mucosal blood flow of the stomach and significantly decreased inflammatory factors and oxidative stress markers. Moreover, BGF significantly reduced the expression of p-NF-κB p65. Consistently, BGF demonstrated similar effects on LPS-induced RAW 264.7 cells as it did in vivo. Conclusion: BGF could accelerate the healing of gastric injury by exerting antioxidant and anti-inflammatory effects and maintaining mucosal integrity.

Determination of Dapoxetine Hydrochloride in Human Plasma by HPLC-MS/MS and Its Application in a Bioequivalence Study.

Dapoxetine is used for the treatment of premature ejaculation. The present study developed an HPLC-MS/MS method to determine the levels of dapoxetine in human plasma processed using simple protein precipitation. Dapoxetine-d7 was selected as the internal standard. The established method was performed using a mass spectrometer equipped with an electrospray ionization source in multiple positive ion reactions to monitor the mode using the precursor-to-product ion transitions of m/z 306.2-157.2 and m/z 313.2-164.2 for dapoxetine-d7 and dapoxetine, respectively. The method was evaluated based on its selectivity, linearity, limit of quantification, precision, accuracy, matrix effects, dilution integrity, stability, and extraction recovery. As a result of the model used in the present study, the validated linear ranges of dapoxetine were determined to be 2.00~1000 ng/mL in plasma, and the selectivity, precision, accuracy, dilution integrity, stability, and extraction recovery met the accepted standard. No matrix interference was observed. The method was successfully validated and applied to pharmacokinetic studies in healthy Chinese volunteers during the fasting and postprandial periods, respectively.

Lupeol inhibits the proliferation and migration of MDA-MB-231 breast cancer cells via a novel crosstalk mechanism between autophagy and the EMT.

Triple-negative breast cancer is the most aggressive type of breast cancer, with a poor prognosis, while effective treatment options are limited. In this study, the anti-tumor effect of lupeol, a natural triterpenoid, toward breast cancer cells and the underlying mechanisms were examined. We firstly predict the primary pathways of lupeol inhibited to TNBC by a network pharmacology approach, which indicated that lupeol may inhibit TNBC via multiple signaling pathways. In addition, experimental data showed that lupeol exhibited outstanding anti-proliferative and anti-metastatic abilities in vitro and in vivo. Additional intrinsic mechanism studies revealed that lupeol might induce autophagy by inhibiting the Akt-mTOR pathway, and activating an autophagy inhibited epithelial-mesenchymal transition (EMT). This study demonstrated that lupeol could inhibit TNBC cells by inducing autophagy, suggesting lupeol as a potential treatment alternative or as a dietary supplement for TNBC, as well as offering novel insights into the anti-cancer effect of lupeol.

Metabolomics and integrated network pharmacology analysis reveal attenuates cardiac hypertrophic mechanisms of HuoXin pill.

ETHNOPHARMACOLOGICAL RELEVANCE: Cardiac hypertrophy (CH) is maladaptive and contributes to the pathogenesis of heart failure. Huoxin pill (HXP), a Chinese herbal prescription, is widely applied in the treatment of cardiovascular disease (CAD). Its mechanism, however, is unclear. AIM OF THE STUDY: This study investigated the mechanism of action for Huoxin pill in the treatment of CH, an important stage of CAD. MATERIALS AND METHODS: A total of 60 rats were injected with isoprenaline (ISO) to establish a model of CH. Echocardiography and histopathologic evaluation were performed to evaluate the disease severity, whereas ELISAs were conducted to determine the expression of oxidative stress. Network pharmacology and metabolomic analyses were conducted to identify the key compounds, core targets and pathways that mediate the effects of HXP against CH. Western blotting and immunohistochemistry were used to test apoptosis protein levels. RESULTS: HXP administration in ISO-treated rats decreased hypertrophy indices, alleviated cardiac pathological damage, and downregulated oxidative stress levels when compared to those of rats subjected to ISO treatment only. Moreover, network pharmacology results suggested that the PI3K-Akt pathway is a main mechanism by which HXP inhibits cardiac hypertrophy, and experimental verification showed that HXP inhibited cardiomyocyte apoptosis via activation of the PI3K-Akt pathway. The results of metabolomic analysis identified 21 differential metabolites between the HXPH group and ISO group, which were considered to be metabolic biomarkers of HXP in the treatment of CH. Among them, 6 differential metabolites were significantly upregulated, and 15 were significantly downregulated. CONCLUSIONS: The present study presents an integrated strategy for investigating the mechanisms of HXP in the treatment of CH and sheds new light on the application of HXP as a traditional Chinese medicine.