Cichoric acid targets RANKL to inhibit osteoclastogenesis and prevent ovariectomy-induced bone loss.

BACKGROUND AND AIM: Osteoporosis, a systemic metabolic bone disease, is characterized by the decline of bone mass and quality due to excessive osteoclast activity. Currently, drug-targeting osteoclasts show promising therapy for osteoporosis. In this study, we investigated the effect of cichoric acid (CA) on receptor activator of nuclear kappa-B ligand (RANKL)-induced osteoclastogenesis and the bone loss induced by ovariectomy in mice. EXPERIMENTAL PROCEDURE: Molecular docking technologies were employed to examine the interaction between CA and RANKL. CCK8 assay was used to evaluate the cell viability under CA treatment. TRAcP staining, podosome belt staining, and bone resorption assays were used to test the effect of CA on osteoclastogenesis and osteoclast function. Further, an OVX-induced osteoporosis mice model was employed to identify the effect of CA on bone loss using micro-CT scanning and histological examination. To investigate underlying mechanisms, network pharmacology was applied to predict the downstream signaling pathways, which were verified by Western blot and immunofluorescence staining. KEY RESULTS: The molecular docking analysis revealed that CA exhibited a specific binding affinity to RANKL, engaging multiple binding sites. CA inhibited RANKL-induced osteoclastogenesis and bone resorption without cytotoxic effects. Mechanistically, CA suppressed RANKL-induced intracellular reactive oxygen species, nuclear factor-kappa B, and mitogen-activated protein kinase pathways, followed by abrogated nuclear factor activated T-cells 1 activity. Consistent with this finding, CA attenuated post-ovariectomy-induced osteoporosis by ameliorating osteoclastogenesis. CONCLUSIONS AND IMPLICATIONS: CA inhibited osteoclast activity and bone loss by targeting RANKL. CA might represent a promising candidate for treating osteoclast-related diseases, such as osteoporosis.

Study of the volatile constituents in radix flemingiae macrophyllae and a substitute by gas chromatography-mass spectrometry and chemometric methods.

A combined approach of subwindow factor analysis and spectral correlative chromatography was used to analyze the volatile components in Radix Flemingiae Macrophyllae and Flemingiae Latifolia Benth, one of its substitutes. After extraction by a water distillation method, the volatile components in Radix Flemingiae Macrophyllae and Flemingiae Latifolia Benth were detected by GC-MS. Then the qualitative and quantitative analysis of the volatile components in Radix Flemingiae Macrophyllae was completed with the help of subwindow factor analysis resolving two-dimensional original data into mass spectra and chromatograms. Sixty five of 82 separated constituents in the total ion chromatogram of the volatile components in Radix Flemingiae Macrophyllae were identified and quantified, accounting for about 88.79% of the total content. Then, spectral correlative chromatography was used to extract correlative constituents in Flemingiae Latifolia Benth. Fifty one correlative components were recognized in essential oil of Flemingiae Latifolia Benth. The result proves the combined approach is powerful in the analysis of complex herbal samples. The developed method can be used to compare the sameness and differences of Radix Flemingiae Macrophyllae and its substitutes and it can also be used for quality control of Radix Flemingiae Macrophyllae.

[Study on the inhibitory effect of Reduqing Oral Liquid on respiratory syncytial virus in vitro].

OBJECTIVE: To explore the antiviral effect of Reduqing Oral Liquid on respiratory syncytial virus (RSV). METHODS: The anti-RSV effect of Reduqing Oral Liquid in Hep-2 cell culture was investigated by the cytopathic effect (CPE) inhibition assay and MTT assay, with ribavirin as the positive control drug. RESULTS: Reduqing Oral Liquid was effectively antiviral agent for RSV in a concentration-dependent manner in vitro. The median toxic concentration (TC50) was 9182.4 mg/L, the median effective concentration (EC50) was 559.8 mg/L and the treatment index (TI) was 16.4. CONCLUSION: Reduqing Oral Liquid has obvious inhibitory effect on RSV in vitro.

[Antiviral effects of an effective section of a prescription of traditional Chinese medicine on influenza virus A in vitro].

OBJECTIVE: To study the inhibitive effects of an effective section of a prescription of traditional Chinese medicine (TCM-ES) on influenza virus A FM1 strain in vitro. METHODS: The experiments were performed by microcytopathic-inhibiting-assay, Neutral Red stain and inhibiting plaque-forming units (PFU) test on MDCK cell strain. By means of observing the cytopathic effects (CPE), measuring the absorbance [D(lambda)] and counting the PFU, according to Reed-Muench assay, the TCM-ES's effective dosage of 50 percentage (EC50) and treatment index (TI) to FM1 were calculated. The inhibiting dose of 50 percentage of PFU (IC50) was also figured up. RESULTS: By CPE assay, TCM-ES'S EC50, MTC and TI to 100TCID50 FM1 strain infection were (300 +/- 18.3) mg/L, (75 +/- 6.8) mg/L and (7.1 +/- 0.7), respectively; Whereas, ribavirin's EC50, MTC and TI was (52.3 +/- 10.1) mg/L, (25 +/- 4.1) mg/L and (20.8 +/- 5.1), respectively. By Neutral Red stain assay,TCM-ES's IC50 and TI was (285.0 +/- 19.2) mg/L and (7.2 +/- 0.6), respectively; whereas ribavirin's IC50 and TI was (45.3 +/- 4. 9) mg/L and (21.2 +/- 3.1), respectively. By reducing PFU assay, the IC50 of TCM-ES and ribavirin was 300 mg/L and 50 mg/L, respectively. All the results above were almost consistent with each other (P > 0.05). CONCLUSIONS: TCM-ES assumes antiviral action on IFV-FM1 strain in a certain degree in vitro and can rebel intracellular virus. But it is worse than the positive control medicine of ribavirin and is worthy of further study.