1.Effects of Bortezomib on proliferation, cell cycle and activation of NF-?B of non-small cell lung cancer cells
Medical Journal of Chinese People's Liberation Army 1983;0(05):-
Objective To study the effect of Bortezomib on proliferation, apoptosis, cell cycles and activation of NF-?B of non-small cell lung cancer cells (NSCLC) in vitro. Methods The inhibitory action of Bortezomib on cellular growth was determined by MTT. The effects of Bortezomib on cell cycle and apoptosis were assessed by flow cytometry. The influence of Bortezomib on the expressions of NF-?B, I?B and Bcl-2 were detected with Western blotting. Results The inhibitory effects of Bortezomib on the proliferation of NSCLC cells showed a time-and concentration-dependent manner. The growth of NSCLC cells was arrested at G2/M stage after treatment with Bortesomib at 25nmol/L for 48h. Basal expression of NF-?B was found to exist in all the 6 cell lines, with NF-?B expression in nucleus showing an inverse correlation with I?B expression in cytoplasm. Bortezomib threw no significant influence on the basal expression of NF-?B, but significantly blocked the TNF-?-induced nuclear translocation of NF-?B and down-regulated the expression of anti-apoptosis protein Bcl-2 in a time-and concentration-dependent manner. Conclusion With NF-?B-dependent pathway, Bortezomib may inhibit the proliferation of NSCLC cells and induce apoptosis.
2.Simultaneous Determination of Liquiritin and Glycyrrhizic Acid in Erxieting Granule by HPLC
Qingxia CHU ; Haijun QIN ; Junling LIU ; Hui ZHANG ; Bo YU ; Yuhan MA ; Yazhong ZHANG
China Pharmacist 2017;20(4):743-745
Objective:To establish an HPLC method for the simultaneous determination of liquiritin and glycyrrhizic acid in Erxieting granule.Methods:A TechMate C18-ST(250 mm×4.6 mm,5 μm) column with a DAD detector was used.The mobile phase consisted of acetonitrile (A) and 0.05% phosphoric acids in water (B) with gradient elution.The flow rate was 1.0 ml·min-1 and the detection wavelength was 237 nm.The sample size was 5 μl and the column temperation was room temperatence.Results:Linear calibration curves were obtained within the range of 10.32-51.62 mg·L-1 for liquiritin and 79.40-397.00 mg·L-1for glycyrrhizic acid.The average spiked recovery of liquiritin and glycyrrhizic acid was 98.10(RSD=1.0%,n=6)and 97.15(RSD=1.8%,n=6),respectively.Conclusion:The method is accurate,reproducible and stable,and can be used for the quality control of Erxieting granule.
3.Mechanism of Flemiphilippinin D Regulating Inflammatory Response in CIA Rats Through TLR2/MyD88/NF-κB Signaling Pathway
Qingxia QIN ; Lianhua HE ; Mei WEI ; Huan HE ; Liuhui MA ; Ting PANG ; Lan ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(17):134-141
ObjectiveTo observe the effect of Flemiphilippinin D on collagen-induced arthritis (CIA) in rats and explore its mechanism. MethodForty rats were randomly divided into normal group, CIA group, methotrexate (MTX) group (1.35 mg·kg-1), low-dose Flemiphilippinin D group (1.5 mg·kg-1), and high-dose Flemiphilippinin D group (3.0 mg·kg-1), with eight rats in each group. Except for the normal group, the CIA model was induced by type Ⅱ collagen. Each group was given corresponding liquid medicine or normal saline, once a week in the MTX group, and once a day in the Flemiphilippinin D groups for a total of 28 days. The arthritis score and joint swelling degree of rats were experimentally recorded. Pathological changes in the ankle joint of rats were observed by hematoxylin-eosin (HE) staining. Serum levels of inflammatory cytokines interleukin (IL)-1β, IL-6, IL-8, and tumor necrosis factor (TNF)-α were detected by enzyme-linked immunoabsorbent assay (ELISA), and the mRNA expression of Toll-like receptor 2 (TLR2), myeloid differentiation factor 88 (MyD88), and nuclear transcription factor-κB (NF-κB) p65 were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR), and the protein expressions of TLR2, MyD88, and NF-κB p65 were detected by Western blot. ResultCompared with the normal group, the ankle joint of the CIA group was significantly swollen, and the clinical score of arthritis and the degree of joint swelling were significantly increased (P<0.01). The ankle joint tissue structure was significantly damaged, and the levels of inflammatory factors IL-1β, IL-6, IL-8, and TNF-α in serum were significantly increased (P<0.01). The mRNA levels and protein levels of TLR2, MyD88, and NF-κB p65 were significantly increased(P<0.01). Compared with the CIA group, arthritis clinical score and joint swelling of rats in each administration group were significantly reduced (P<0.05, P<0.01), and the pathological changes in the ankle joint were significantly improved. The contents of serum IL-1β, IL-6, IL-8, and TNF-α were significantly decreased (P<0.05, P<0.01). The mRNA levels and protein levels of TLR2, MyD88, and NF-κB p65 in the ankle joint were significantly decreased (P<0.05, P<0.01). ConclusionTo a certain extent, Flemiphilippinin D can reduce the expression of inflammatory factors in rheumatoid arthritis rats and play a good therapeutic effect. It works perhaps by inhibiting the activation of the TLR2/MyD88/NF-κB signaling pathway and thus shows an anti-inflammatory effect.