Glechomenes A-G, diterpenoids with anti-inflammatory activities from the aerial part of Glechoma longituba.

Ten diterpenoids including six unreported abietane-type diterpenoids Glecholmenes A-F (1-6) and an undescribed labdane-type diterpenoid Glecholmene G (9), together with three known diterpenoids (7,8,10), were firstly isolated from the aerial part of G. longituba. Their structures were established mainly by nuclear magnetic resonance (NMR) and high-resolution electrospray ionization mass spectrometry (HR-ESI-MS) methods. Electronic circular dichroism (ECD) calculations and X-ray crystallographic analyses were used for the determination of their absolute configurations. The anti-inflammatory activity of all compounds was evaluated using the classical LPS-induced NO release model in RAW264.7 cells. Compound 2 displayed significant anti-inflammatory activities with IC50 values of 29.08 ± 1.40 µM (Aminoguanidine hydrochloride as the positive control, IC50 = 21.84 ± 0.48 µM).

Six polyacetylenes from Atractylodes macrocephala Koidz and their anti-colon cancer activity.

Six undescribed polyacetylenes Atracetylenes A-F (1-6) and three known ones (7-9) were isolated from the rhizomes of Atractylodes macrocephala Koidz.. The comprehensive interpretation of NMR, HR-ESI-MS, DP4+ calculations, and electronic circular dichroism (ECD) calculations resulted in the elucidation of their structures and absolute configurations. The anti-colon cancer activities of (1-9) were evaluated by assaying the cytotoxicity and apoptosis on CT-26 cell lines. Notably, 5 (IC50 17.51 ± 1.41 µM) and 7 (IC50 18.58 ± 1.37 µM) exhibited significant cytotoxicity, and polyacetylenes 3-6 showed excellent abilities to promote apoptosis of CT-26 cell lines by Annexin V-FITC/PI assay. The results demonstrated that the polyacetylenes in A. macrocephala may be prospective for the treatment of colorectal cancer.

Pentacyclic triterpenoids from spikes of Prunella vulgaris L. with thyroid tumour cell cytostatic bioactivities.

Five new triterpenoids, including four ursane types (1-4) and one oleanane type (5), together with 15 known ursane types pentacyclic triterpenoids (6-20) were isolated from the fruit spikes of Prunella vulgaris L., a traditional Chinese herbal medicine. Their structures were elucidated based on IR, HR-ESI-MS, and NMR spectroscopic data. The SW579 cell line was used to evaluate anti-thyroid cancer activities of (1-20). The results indicated that (7-9), (16), and (19) exhibited apparent inhibitory activity with IC50 values of 25.73-71.41 µM (cisplatin as positive control, IC50 14.49 ± 0.97 µM). Network pharmacology and molecular docking were also used for the prediction of the synergistic actions and the underlying mechanisms. Accordingly, four potential targets have been characterized.

Kaempferol impairs aerobic glycolysis against melanoma metastasis via inhibiting the mitochondrial binding of HK2 and VDAC1.

Metastasis is the leading cause of death in melanoma patients. Aerobic glycolysis is a common metabolic feature in tumor and is closely related to cell growth and metastasis. Kaempferol (KAM) is one of the active ingredients in the total flavonoids of Chinese traditional medicine Sparganii Rhizoma. Studies have shown that it interferes with the cell cycle, apoptosis, angiogenesis and metastasis of tumor cells, but whether it can affect the aerobic glycolysis of melanoma is still unclear. Here, we explored the effects and mechanisms of KAM on melanoma metastasis and aerobic glycolysis. KAM inhibited the migration and invasion of A375 and B16F10 cells, and reduced the lung metastasis of melanoma cells. Extracellular acidification rates (ECAR) and glucose consumption were obviously suppressed by KAM, as well as the production of ATP, pyruvate and lactate. Mechanistically, the activity of hexokinase (HK), the first key kinase of aerobic glycolysis, was significantly inhibited by KAM. Although the total protein expression of HK2 was not significantly changed, the binding of HK2 and voltage-dependent anion channel 1 (VDAC1) on mitochondria was inhibited by KAM through AKT/GSK-3ß signal pathway. In conclusion, KAM inhibits melanoma metastasis via blocking aerobic glycolysis of melanoma cells, in which the binding of HK2 and VDAC1 on mitochondria was broken.

Hypoxia Boosts Aerobic Glycolysis in Carcinoma: A Complex Process for Tumour Development.

Hypoxia, a common feature of malignant tumours, is mainly caused by insufficient oxygen supply. Hypoxia is closely associated with cancer development, affecting cancer invasion, metastasis, energy metabolism, and other pathological processes, and is not conducive to cancer treatment and prognosis. Tumour cells exacerbate metabolic abnormalities to adapt to the hypoxic microenvironment, especially to enhance aerobic glycolysis. Glycolysis leads to an acidic microenvironment in cancer tissues, enhancing cancer metastasis, deterioration, and drug resistance. Therefore, hypoxia is a therapeutic target that cannot be ignored in cancer treatment. The adaptation of tumour cells to hypoxia is mainly regulated by hypoxia-inducible factors (HIFs), and the stability of HIFs is improved under hypoxic conditions. HIFs can promote glycolysis in tumours by regulating glycolytic enzymes, transporters, and the TCA cycle. In addition, HIFs indirectly affect glycolysis by interacting with non- coding RNAs. Therefore, targeting hypoxia and HIFs is important for tumour therapy.

FBXO22, ubiquitination degradation of PHLPP1, ameliorates rotenone induced neurotoxicity by activating AKT pathway.

Parkinson's disease (PD) is a neurodegenerative disease caused by the lacking of dopaminergic neurons. Many reports have illustrated that rotenone is applied to establish the experimental model of PD, which simulates PD-like symptoms. FBXO22 is a poorly understood protein that may be involved in neurological disorders. However, little is known about FBXO22 in PD. In this study, first, SH-SY5Y cells were treated with rotenone to construct PD model in vitro. It was discovered that the FBXO22 expression was down-regulated following rotenone treatment. Additionally, overexpression of FBXO22 reduced rotenone treatment-mediated cell apoptosis in SH-SY5Y cells. In view of the ubiquitination effect of FBXO22, our study uncovered that FBXO22 bound with and degraded PHLPP1 by ubiquitination. Next, the effects of PHLPP1 on AKT pathway in PD were further explored. It was demonstrated that PHLPP1 inactivated AKT pathway through down-regulating the pAKT/AKT and pmTOR/mTOR levels. Through rescue assays, the results showed that PHLPP1 overexpression partially reversed the reduction of rotenone induced neurotoxicity caused by FBXO22 overexpression. Finally, we found that overexpression of FBXO22 alleviated rotenone-induced PD symptoms in rat model. Moreover, it was discovered that l-dopa treatment could not affect the FBXO22 expression in PD. In conclusion, findings from our work proved that FBXO22 degraded PHLPP1 by ubiquitination to ameliorate rotenone induced neurotoxicity, which attributed to activate AKT pathway. This work suggested that FBXO22 may be an effective biological marker for PD treatment.

Cryptotanshinone Inhibits ERα-Dependent and -Independent BCRP Oligomer Formation to Reverse Multidrug Resistance in Breast Cancer.

Both long-term anti-estrogen therapy and estrogen receptor-negative breast cancer contribute to drug resistance, causing poor prognosis in breast cancer patients. Breast cancer resistance protein (BCRP) plays an important role in multidrug resistance. Here, we show that cryptotanshinone (CPT), an anti-estrogen compound, inhibited the oligomer formation of BCRP on the cell membrane, thus blocking its efflux function. The inhibitory effect of CPT on BCRP was dependent on the expression level of estrogen receptor α (ERα) in ERα-positive breast cancer cells. Furthermore, ERα-negative breast cancer cells with high expression of BCRP were also sensitive to CPT because CPT was able to bind to BCRP and inhibit its oligomer formation on the cell membrane, suggesting that the high level of BCRP expression is crucial for CPT to reverse drug resistance. The combination of CPT and chemotherapeutic agents displayed enhanced anticancer effects. The results suggest that CPT is a novel BCRP inhibitor via blocking the oligomer formation of BCRP on the cell membrane. CPT is able to inhibit the activity of BCRP in an ERα-dependent and -independent manner, sensitizing breast cancer cells to chemotherapy.

Repetitive transcranial magnetic stimulation in combination with neuromuscular electrical stimulation for treatment of post-stroke dysphagia.

OBJECTIVE: This study was performed to determine whether repetitive transcranial magnetic stimulation (rTMS) combined with neuromuscular electrical stimulation (NMES) effectively ameliorates dysphagia and how rTMS protocols (bilateral vs. unilateral) combined with NMES can be optimized. METHODS: Sixty-four patients were randomly divided into four groups using a random distribution table: the sham rTMS plus NMES (Sham-rTMS/NMES), ipsilesional 10-Hz rTMS plus NMES (Ipsi-rTMS/NMES), contralesional 1-Hz rTMS plus NMES (Contra-rTMS/NMES), and bilateral rTMS plus NMES (Bi-rTMS/NMES) groups. Cortical excitability as measured by the amplitude of the motor evoked potential at the mylohyoid muscle cortical representative area, swallowing function as measured by the Standardized Swallowing Assessment, and the degree of dysphagia were evaluated at baseline, after the stimulation course, and at the 1-month follow-up. RESULTS: Bi-rTMS/NMES produced higher cortical excitability and better swallowing function recovery. Compared with NMES alone, unilateral rTMS plus NMES had additional effects on cortical excitability and rehabilitation of dysphagia, but there were no differences between the Contra-rTMS/NMES and Ipsi-rTMS/NMES groups. No adverse events occurred. CONCLUSION: The combination of rTMS with NMES was superior to NMES alone in improving the recovery of post-stroke dysphagia, and the combination of bilateral rTMS with NMES was more effective than unilateral rTMS combined with NMES.

[Effects of N-acetylcysteine upon methylglyoxal-induced damage in hippocampal neuronal cells].

OBJECTIVE: To explore the effects of N-acetylcysteine (NAC) upon the methylglyoxal (MG)-induced injury of hippocampal neuronal cells. METHODS: Primary cultures of 1-day-old SD rat hippocampal neuron were exposed to MG and/or NAC for 24 h respectively. Apoptosis was quantified by flow cytometer using annexin V-FITC and propidium iodide (PI) staining. The level of intracellular reactive oxygen species (ROS) was measured by an oxidant-sensitive dye 2,7-dichlorofluorescin diacetate (DCFH). The protein and mRNA levels of BDNF and TrkB were assayed with Western blot and real-time reverse-transcription polymerase chain reaction (RT-PCR) respectively. RESULTS: After a 24 h cell incubation with 100 micromol/L MG, the ratio of apoptotic cells in MG group (8.80 +/- 0.31)% significantly increased versus the control group (1.60 +/- 0.15)% and MG+NAC group (4.83 +/- 0.31)% respectively. The level of intracellular oxidation of MG group (10 229 +/- 946) also significantly increased versus the NAC group (2118 +/- 320), control group (4265 +/- 82), MG + NAC group (3886 +/- 415) and pretreated NAC + MG group (2997 +/- 606). MG increased the cellular levels of BDNF but decreased the TrkB mRNA and protein expression significantly. However, NAC significantly decreased the BDNF and increased the TrkB mRNA and protein expression in rat hippocampal neuron after MG induction. All these differences were considered statistically significant. CONCLUSION: MG-induced neurotoxicity in hippocampal neurons is mediated by oxidative stress and it can impair the BDNF/TrkB signal pathway. Antioxidant NAC has protective effect upon MG-induced neurotoxicity through its antiapoptotic action and its antioxidant effect on ROS level. And it works partly by activating the BDNF/TrkB signal pathway.