1.Determination of pueratin in Jiangzhi Hugan capsule by HPLC
Wei ZHENG ; Dongjie NI ; Qing WEI ; Hui XIANG ; Lanlan HU ; Liang SUN
Journal of Pharmaceutical Practice 2017;35(3):259-261
Objective To develop a HPLC method for determination of pueratin.Methods The separation was carried out on a Waters Symmetry C18 column(4.6 mm×250 mm, 5 μm), the mobile phase was composed of acetonitrile and 1% formic acid(11∶89), the detection wavelength was set at 250 nm, the flow rate was 1.0 ml/min, the column temperature was 30 ℃ and the injection volume was 10 μl.Results The linearity was obtained over 2~40 μg/ml (r=0.999 8) for pueratin.The RSD of precision were less than 2%.The average recovery was between 98% and 103%.Conclusion This HPLC method was simple, accuracy and suitable for the quality control of Jiangzhi Hugan capsule.
2.Sensitized cytotoxic effects of dexamethasone or radiation on multiple myeloma cells by chloroquine
Hui FU ; Yulong TAO ; Min NI ; Dongjie LI ; Fuming SHEN
Journal of Pharmaceutical Practice 2016;34(3):210-214
Objective To investigate the role and possible mechanism of combination use of chloroquine (CQ) with either dexamethasone (DEX) or radiation on multiple myeloma (MM) cell line U266 .Methods Cell viability of U266 treated with CQ alone ,or CQ combined with either DEX or radiation was measured by cell counting kit-8 (cck8) .CalcuSyn method was used to assess effect of drugs interaction .Cell viability and apoptosis of U266 pre-treated with CQ were also measured by cck8 and flow cytometry after radiation .Expression of B-cellymphoma-2 (Bcl-2) in U266 cells treated by CQ combined with DEX or radiation was determined by Western blot analysis .Results Either CQ or DEX displayed a dose dependent cell proliferation in-hibitory effect on U266 cells .Cytotoxic effect of DEX (125 μmol/L) on U266 cells was enhanced and expression of Bcl-2 pro-tein in U266 cells was decreased by combining with CQ (3 .9 μmol/L) .U266 cells were sensitized to radiation and cell death was induced by CQ (1 .0 μmol/L) .Conclusion CQ could sensitize cytotoxic effect of DEX or radiation on U 266 cells ,and the former was possibly related to down-regulation of Bcl-2 protein .
3.One step assay of the four plant ingredients in Yangxue Anshen syrup by HPLC method
Ping LU ; Dongjie NI ; Wei ZHENG ; Liangjun GUO ; Xiang WANG
Journal of Pharmaceutical Practice 2021;39(3):270-273
Objective To establish a HPLC method for simultaneous determination of quercitrin, luteoloside, rutin and 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside in Yangxue Anshen syrup. Methods Waters symmetry C18 column (250 mm×4.6 mm, 5 μm) was used with 0.1% acetic acid (A) and methanol (B) as the mobile phase. Gradient elution was performed at a flow rate of 1.0 ml/min, 0-15 min, 95%-90%A; 15-35 min, 90%-70%A; 35-55 min, 70%-60%A; 55-85 min, 60%-50%A; 85-95 min, 10%A. The detection wavelengths were 256 nm and 320 nm. Column temperature was 30 ℃ and the injection volume was 10 μl. Results Quercitrin, luteoloside, rutin and 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside showed good linear relationship within the range of 10-300, 5.0-150.0, 5.0-150.0, 20.0-600.0 µg/ml(r≥0.9989), respectively. The average recovery was (96.75±1.41)%, (99.61±1.01)%, (97.18±1.96)% and(99.12±0.97)% (n=6), respectively. Conclusion The established method is simple, accurate and stable, which can be used for the simultaneous determination of 4 components in Yangxue Anshen syrup.
4.Structures of the portal vertex reveal essential protein-protein interactions for Herpesvirus assembly and maturation.
Nan WANG ; Wenyuan CHEN ; Ling ZHU ; Dongjie ZHU ; Rui FENG ; Jialing WANG ; Bin ZHU ; Xinzheng ZHANG ; Xiaoqing CHEN ; Xianjie LIU ; Runbin YAN ; Dongyao NI ; Grace Guoying ZHOU ; Hongrong LIU ; Zihe RAO ; Xiangxi WANG
Protein & Cell 2020;11(5):366-373
5.Targeting a novel inducible GPX4 alternative isoform to alleviate ferroptosis and treat metabolic-associated fatty liver disease.
Jie TONG ; Dongjie LI ; Hongbo MENG ; Diyang SUN ; Xiuting LAN ; Min NI ; Jiawei MA ; Feiyan ZENG ; Sijia SUN ; Jiangtao FU ; Guoqiang LI ; Qingxin JI ; Guoyan ZHANG ; Qirui SHEN ; Yuanyuan WANG ; Jiahui ZHU ; Yi ZHAO ; Xujie WANG ; Yi LIU ; Shenxi OUYANG ; Chunquan SHENG ; Fuming SHEN ; Pei WANG
Acta Pharmaceutica Sinica B 2022;12(9):3650-3666
Metabolic-associated fatty liver disease (MAFLD), which is previously known as non-alcoholic fatty liver disease (NAFLD), represents a major health concern worldwide with limited therapy. Here, we provide evidence that ferroptosis, a novel form of regulated cell death characterized by iron-driven lipid peroxidation, was comprehensively activated in liver tissues from MAFLD patients. The canonical-GPX4 (cGPX4), which is the most important negative controller of ferroptosis, is downregulated at protein but not mRNA level. Interestingly, a non-canonical GPX4 transcript-variant is induced (inducible-GPX4, iGPX4) in MAFLD condition. The high fat-fructose/sucrose diet (HFFD) and methionine/choline-deficient diet (MCD)-induced MAFLD pathologies, including hepatocellular ballooning, steatohepatitis and fibrosis, were attenuated and aggravated, respectively, in cGPX4-and iGPX4-knockin mice. cGPX4 and iGPX4 isoforms also displayed opposing effects on oxidative stress and ferroptosis in hepatocytes. Knockdown of iGPX4 by siRNA alleviated lipid stress, ferroptosis and cell injury. Mechanistically, the triggered iGPX4 interacts with cGPX4 to facilitate the transformation of cGPX4 from enzymatic-active monomer to enzymatic-inactive oligomers upon lipid stress, and thus promotes ferroptosis. Co-immunoprecipitation and nano LC-MS/MS analyses confirmed the interaction between iGPX4 and cGPX4. Our results reveal a detrimental role of non-canonical GPX4 isoform in ferroptosis, and indicate selectively targeting iGPX4 may be a promising therapeutic strategy for MAFLD.