1.Astragalus prevents photoreceptor cell apoptosis induced by N-methyl-N-nitrosourea in rats
Jinnan YANG ; Taizhen CUI ; Ying ZHAO ; Shengying LI ; Heqin ZHAN
Chinese Journal of Tissue Engineering Research 2006;10(39):177-179
BACKGROUND: Retinitis pigmentosa (RP) is a non-inflammatory, bilaterally progressive, retinal degeneration characterized by loss of photoreceptor cells via an apoptotic mechanism, and it eventually leads to blindness.Research shows that the traditional Chinese medicines of Astragalus has great prospect on blocking the progression of RP disease.OBJECTIVE: To observe the protective effect of Astragalus on N-methylN-nitrosourea (MNU)-induced retinal damage in Sprague-Dawley (SD) rats and provide the optimal treatment for RP in humans.DESIGN: Randomized controlled experiment.SETTING: School of Pharmaceutical Sciences, Xinxiang Medical College.MATERIALS :The experiment was completed in Pharmacological Laboratory of Zhongshan Ophthalmic Centre, Sun Yat-sen University between March to December 2004. Totally 114 female SD rats were purchased from the Animal Center of Zhongshan Medical College, Sun Yat-sen University.MNU was purchased from Sigma Company of America. Astragalus injection was purchased from Chengdu Diao Jiuhong Pharmaceutical Factory (Batch No. Z99060535, 2 mL/ampoule, main ingredient: Astragalus).METHODS: Among 114rats, 30 were for morphometric analysis of retinal layers, 30 were for detection of apoptosis and 54 were for detection of NF-κB p65 activity. All of them were randomly divided into different groups and each group had 6 rats. Astragalus at doses of 2.5, 5 and 10 g/kg were injected intraperitoneally into 47-day rats once a day. Meanwhile, a single intraperitoneal injection of 60 mg/kg MNU was given to 50-day rats in model and Astragalus groups. At different intervals after MNU treatment,the animals were sacrificed. Retinal damage was evaluated based on retinal thickness, the apoptotic index of the photoreceptor cells was detected by TUNEL labeling and the DNA-binding activity of NF-κB p65 was analyzed according to transcription factor assay kit.MAIN OUTCOME MEASURES: Comparison of retinal thickness, apoptotic index and the activity of nuclear NF-κB p65.RESULTS: Totally 114 rats entered the result analysis. Pretreatment with Astragalus could dose-dependently suppress MNU-induced photoreceptor cell loss and decreased the apoptotic index. Astragalus at dose of 10 g/kg also time-dependently up-regulated the activity of nuclear NF-κB p65.However, protective effect of Astragalus on MNU-induced central retinal damage was not found.CONCLUSION: Astragalus partially protects against MNU-induced retinal damage by up-modulating the activity of nuclear NF-κB p65 to inhibit apoptosis of photoreceptor cells in a dose-dependent manner.
2.Chemical constituents in rhizome of Pterocypsela elata and activity of lactuside B against brain ischemia
Heqin ZHAN ; Lanqing GUO ; Jianmin CUI ; Fulin YAN
Chinese Traditional and Herbal Drugs 1994;0(05):-
Objective To investigate the major chemical composition in rhizome of Pterocypsela elata and explore the activity of lactuside B against brain ischemia.Methods The compounds were isolated and purified by silica gel column chromatography and elucidated by spectroscopic experiments;The model of partial brain ischemia was used to detect water content,MDA and SOD levels in brain tissue in order to observe the activity of lactuside B against brain ischemia.Results Ten compounds were obtained and established as lactuside B(1),11?,13-dihydrolactucin acetate(2),?-sitosterol(3),daucosterol(4),(24R)-5?-stigrnast-7,22(E)-dien-3?-ol(5),3,3',4-trimethoxylellagic acid(6),?-amyrin(7),oleanolic acid(8),n-hexacosanic acid(9),and stearic acid(10).Lactuside B was a key component,and it's yield was 0.15%.Contents of water and MDA level in the brain tissue were significantly decreased,and the SOD content notably increased in all groups of lactuside B.Conclusion Ten compounds are all isolated from this plant for the first time.Compound 1 is a key component which possesses obvious activity against brain ischemia.
3.Inhibitory effect of miR-205 targeted YES1 on proliferation of A549 cells
Yuan CHENG ; Yongzhan ZHEN ; Xiaofang HAO ; Pengyu WU ; Yanan XIONG ; Zhiyong LIU ; Heqin CUI
Journal of Jilin University(Medicine Edition) 2014;(3):493-498
Objective To detect the expression levels of the miR-205 in lung cancer tissue and A549 cells and its targeted gene YES1 using qRT-PCR and dual fluorescence protein repoter assay system,and to explore the possible mechanism of miR-205 to inhibit the proliferation of lung cancer A549 cells.Methods The expression levels of miR-205 in 10 cases of lung cancer tissue and adjacent normal lung tissue were detected with qRT-PCR.The cell growth curve and colony formation assay were used to determine the proliferation rate of A549 cells after transfected by miR-205 mimics and control mimics.The sequences of YES1 3′UTR (untranslated region)and mutation target sites of YES1 3′UTR were inserted into the plasmid which expressed green fluorescence protein (pcDNA3/EGFP) respectively to construct the green fluorescence protein plasmids of YES1-3′UTR and mut-YES1-3′UTR. There were six groups in the study:YES1-3′UTR, YES1-3′UTR and miR-205 mimics, YES1-3′UTR and control mimics,mut-YES1-3′UTR, mut-YES1-3′UTR and miR-205 mimics, mut-YES1-3′UTR and control mimics;after the plasmids expressed red fluorescent protein (pDsRed2-N1 )were cotransfected into A549 cells,the extracted protein was detected with fluorescence spectrophotometer.Results Compared with adjacent normal lung tissue,the expression levels of miR-205 in lung cancer tissue and A549 cells were decreased (P<0.05 );the proliferation rate of A549 cells in miR-205 mimics group was lower than that in control mimics group (P<0.05). The fluorescence protein expression level in YES1-3′UTR and miR-205 mimics co-transfected group was lower than that in YES1-3′UTR and control mimics co-transfected group, the difference was statistically significant (P<0.01).The number of cell colony formation of A549 cells in highly expressed YES1 group was higher than that in cell control group (P<0.05).Conclusion MiR-205 may inhibit the proliferation of A549 cells through regulating of the expression of YES1 directly.miR-205 and YES1 are potential therapeutic targets for the biological treatment of tumor.