1.HTS model for Poly(ADP-ribose) polymerase-1 inhibitors
Chinese Pharmacological Bulletin 1987;0(01):-
Aim To introduce a high throughput screen model for PARP-1 inhibitors. Methods Setting up an assay for PARP-1 activity relies on the conversion of NAD~+into a highly fluorescent compound. The inhibitory effects of 9 280 samples(including pure organic compounds, extracts from plants and extracts from microorganism)were screened by the high throughout assay. Results 148 compounds had inhibitory effects over 70%. Ultimately, three inhibitors were identified as PARP-1 inhibitor with high activity.Conclusion The high throughput screening was a highly sensitive, inexpensive, and operationally simple assay method in identifying PARP-1 inhibitors.
2.Autophagic Cell Death in Glioma Cell Induced by Ceramide through JNK/c-Jun Pathway
Luyong ZHANG ; Miao ZHANG ; Shibu LIU ; Rui LI
Chinese Journal of Rehabilitation Theory and Practice 2015;21(8):905-912
Objective To observe the autophagy of 87-MG and U251 glioma cells induced by ceramide and explore the possible mechanism. Methods The viability and apoptosis of 87-MG and U251 cells were detected by MTT assay and flow cytometry, respectively. Autophagic- related protein expressions of LC3B /LC3A and Beclin-1 were determined by Western blotting. The activation of JNK/c-Jun signaling pathway induced by ceramide with or without the treatment of JNK specific inhibitor SP600125 was also measured. Results 24 hours after treatment of ceramide, the growth of 87-MG and U251 cells was significantly inhibited time-dependently (P<0.05); and the number of autophagic cells increased dose-dependently (P<0.05). The levels of LC3B/LC3A and Beclin-1 significantly increased after ceramide treatment (P<0.05). JNK signaling pathway was activated in the 87-MG and U251 cells and the phosphorylation of c-Jun also increased after ceramide treatment. This activation of autophagy could be reversed by the pre-treatment of SP600125. Conclusion Ceramide may induce autophagy in 87-MG and U251 glioma cells and the mechanism may be related to the activation of JNK/c-Jun signaling pathway.
3.Effect of antioxidants on lysis of red blood cells induced by saponins
Hongyi WANG ; Boyang YU ; Luyong ZHANG ; Min QIAN
Chinese Traditional and Herbal Drugs 1994;0(07):-
Object To study the hemolytic effect of total saponins from Ginseng (Gs), Radix Ophiopogonis (ROs) and Shengmaisan, a compound preparation of the two saponins, and the antihemolytic effect of antioxidants on hemolysis. Methods The percentage of hemolysis was indirectly calculated by measured cyanomethemoglobin, and morphological changes of red blood cells (RBCs) was examined by light microscope. Results Gs did not lead to hemolysis while both ROs and Shengmaisan induced lysis in a concentration dependent manner. RBCs deformation can be observed by light microscopic study. The hemolysis was partly inhibited by addition of glucose or antioxidants including mannitol and ?-tocopherol. Conclusion The intensity of hemolytic effect varied with different saponins present in the compound preparation, and may be associated with the type of saponin and partly due to its ability to enhance lipid preoxidation of RBCs membrane.
4.Effect and mechanism of Anandamide on the proliferation of human lung cancer A549 cell line
Min CHEN ; Cuifen WANG ; Lixin SUN ; Luyong ZHANG
Chinese Pharmacological Bulletin 1986;0(05):-
Aim The effect of N-arachidonoyl ethanolamide(Anandamide,AEA),on the human lung cancer cell A549 was analysed the possible mechanism was detected.Methods To assess the sensitivity of A549 to AEA,A549 cells were exposed to increasing doses of AEA with or without the antagonists to vanilloid receptor 1(VR1) and aspirin.MTT methods were employed to investigate A549 cell proliferation.Results A549 cells exhibited dose-dependent sensitivity to AEA resulting in dramatic cell death.But the effect of AEA on A549 could not be antagonized by the antagonists such as capsazepine and Ruthenium Red.However,cyclooxygenase(COX) inhibitor,aspirin,could attenuate A549 cell death caused by AEA(P
5.The comparative study of two strains on results of Local lymph node assays
Peili HU ; Luyong ZHANG ; Bo LI ; Shuxia XING
Chinese Journal of Comparative Medicine 2015;(5):54-57
Objective To analyze and compare results of local lymph node assays (LLNA) between BALB/c and CBA mice.Methods 4 chemicals (2,4-dinitrochlorobenzene, eugenol, hexyl cinnamic aldehyde and isopropanol) and 3 cosmetics were applied to the dorsum of both ears of Balb/c and CBA mice for three consecutive days.BrdU solution was injected inter-peritoneally on day 5.On day 6, the bilateral draining auricular lymph nodes were excised and made into single cell suspension, the lymph cell proliferation was measured by BrdU ELISA kit.Results 2,4-dinitrochlorobenzene, eugenol, hexyl cinnamic aldehyde and NO.3 perm agent pretended positive for both strains, EC1.6 values of three chemicals were found to be 0.08% (very strong), 4.02% (moderate), 6.68% (moderate) and 0.07% (very strong), 6.08% ( moderate), 8.89% ( moderate) for BALB/c and CBA mice respectively.Isopropanol, NO.1 and NO.2 cosmetics pretended to be non-sensitizers with SI <1.6 for both strains.Conclusion This study showed that BALB/c mouse was essentially equal to CBA for LLNA: BrdU-ELISA, which suggested that BALB/c mouse was a good alternative for CBA used in chemicals and cosmetics allergenic evaluation.
6.Inhibition of glucometabolism by a novel dehydroabielylamine derivative,DHAA-urea,in human hepatoma HepG2 cells
Jianxiang XIE ; Ling HE ; Luyong ZHANG ; Xiaoping RAO ; Zhanqian SONG
Journal of China Pharmaceutical University 2010;41(2):160-165
The effects of DHAA-urea,a novel dehydroabietylamine(DHAA) derivatives,on cell viability and glucose metabolism,in hypoxia and normoxia human hepatoma HepG2 cells were investigated.Hypoxia cells were achieved using DMEM containing high concentration of glucose without serum and pre-incubating of CoCl_2 (final concentration 150 μmol/L) for 24 h.The antiproliferation effect of DHAA-urea was measured by colorimetric MTT assay.The cellular ATP concentration,the lactate dehydrogenase(LDH) and glucose-6-phosphate dehydro genase (G6PD) activity were detected by their kits.It was shown that DHAA-urea markedly inhibited cell viability,cellular ATP level,LDH and G6PD activity in either aerobic or anaerobic circumstance in a dose-and time dependent manner.This suggested that DHAA-urea possibly inhibited HepG2 cells growth via the inhibition of glucolysis and glucolysis-dependent ATP depletion.DHAA-urea could be a promising candidate in the development of a novel class of agents used for human hepatocellular carcinoma.
7.T Subsets and Antitumor Activity of Lymphocytes Infiltrating Hunan Primary Brain Gliomas
Youjun LI ; Cheng ZHU ; Xiantao KONG ; Guangji ZHANG ; Yumin LIANG ; Luyong ZHANG
Academic Journal of Second Military Medical University 1982;0(01):-
Glioma-infiltrating lymphocytes (GIL) were isolated from 9 surgical biopsy specimens of primary brain gliomas using mechanical and enzymatic digestion and discontinuous density gtadient centrifugation. During cultured in the presence of interieukin-2 (IL-2) for a period of four weeks, GIL were expanded 48.4-fold on the averags, even up to 118-fold. GIL activated by IL-2 had specific cytorytic activity against autologous glioma cells. Analysis of T subsets of GIL freshly isolated showed that CD3+ cells were 71.0?11.9%, CD4+ cells 34.2?6.1% and CD8+ cells 37.0?7.6%. Ability of activated GIL to secrete ?-interferon (?-IFN) was significantly higher than that of freshly isolated GIL and autologous peripheral blood lymphocytes (PBL). The results suggest that GIL have many advantages for an adoptive immunotherapy of patients with brain gliomas and is a new type of antitumor immune effector.
8.Synthesis and biological activity of heterocycle-fused derivatives of pentacylic triterpenes as glycogen phosphorylase inhibitors
Xiaoan WEN ; Yingxia ZHANG ; Jun LIU ; Luyong ZHANG ; Peizhou NI ; Hongbin SUN
Journal of China Pharmaceutical University 2009;40(6):491-496
Aim: To search for novel modulators of glycogen metabolism through structural modifications of natural pentacyclic triterpenes. Methods: A series of N-heterocyclic derivatives were synthesized by fusing indole, qui-noxaline and pyrazine rings with A-ring of oleanolic and ursolic acids. The compounds were biologically evaluated for their inhibitory activity against rabbit muscle glycogen phosphorylase. Results and Conclusion: Twelve heter-ocyclic triterpene derivatives were synthesized and their structures were confirmed by IR, ~1H NMR, ~(13)C NMR and MS. Except for compound 12, all of the compounds exhibited glycogen phosphorylase inhibitory activity with IC_(50) values in the range of 14-252 μmol/L Among this series of compounds, compound 15 showed the best potency with IC_(50), of 14 μmol/L
9.Establishment of M1 Receptor Screening Model for Drugs for Alzheimer's Disease Using Radioimmunoassay Method
Luyong ZHANG ; Zhengzhou JIANG ; Zhaohui CAI ; Xiaochen ZHAO ; Ming YAN ; Xinyan LI
Journal of China Pharmaceutical University 2003;(1):41-45
AIM:To establish a screening model based on muscarinic receptor type 1(M1) for drugs against Alzheimer's disease (AD). METHOD: Human M1 receptor was expressed in HEK293 (human embryonic kidney) cells, and its activation was measured by the intracellular cAMP (cyclic AMP) level. Exogeneous 3H-cAMP was used to compete the intracellular cAMP binding sites. Acetylcholine chloride was used as a positive drug to ensure the sensitivity of this model. RESULT: HEK293 cell expressing system of human M1 receptor was established. Different concentrations of acetylcholine chloride (10-9~10-4 mol/L) activation of M1 receptor leads to an increase of intracellular cAMP 10.343×10-4~33.754×10-4 pmol/μl. CONCLUTION:This screening model has positive response to M1 receptor agonist and can be used for drug screening.
10.Effects of HZ08,a novel P-glycoprotein inhibitor, on the reversal of P-glyco-protein mediated multidrug resistance in nude mice and cytochrome P-450 ac-tivities in rat liver microsomes
Fang YAN ; Yunman LI ; Qiujuan WANG ; Weirong FANG ; Kai KANG ; Luyong ZHANG
Journal of China Pharmaceutical University 2008;(5):447-452
Aim: To evaluate the effects of HZ08, a novel P-glycoprotein inhibitor, on reversing tumor resistance of K562/ADM to adriamycin in nude mice and on the activities of cytochromes P-450 (GYP) isoforms. Methods: Nude mice bearing K562/ADM were injected at different doses of HZ08 with adriamycin for 4 weeks. The tumor weights of HZ08 treatment groups were determined and compared to those of the control and positive groups. In addition, the effects of HZ08 were examined on GYP isoforms-mediated metabolism of specific substrates by GYP isoforms in rat liver microsomes in the presence or absence of HZ08. Results: The tumor weights of HZ08 treatment groups were significantly decreased and HZ08 was a relatively potent inhibitor of CYP3A4, with no significant effects on other isoforms tested. Conclusion: HZ08 has potent effects on reversing P-glycoprotein mediated tumor multidrug resistance in rive with little influence on cytoehrome P-450 activities of rat liver.