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Objecttve To explore the roles of p53,Bax and Bcl-2 in rat lens epithelial cells apoptosis induced by ultraviolet radiation.Methods Healthy SpragueDawley rats (40,6-weeks old,150 g) were selected and divided into 5 groups randomly.After SD rats were injected 100 g · L-1 chloral hydrate (0.35 mL/100 g) intraperitoneally and pupil dilation,the eyes of SD rats were radiated 15 minutes by using UV-B (300-320 nm).The exposed animals were sacrificed at 1 day,3 days,5 days and 7 days after exposure.Both lenses from all animals were extracted.The apoptosis of rats lens epithelial cells was performed by Hoechst 33258 staining.The expression of p53,Bax and Bcl-2 in rat lens epithelial cells at mRNA level was detected by real-time PCR.The distribution and expression of p53,Bax and Bcl-2 were observed by immunohlstochemistry.Results The apoptosis of lens epithelial cells was aggravated after exposure.The expression of p53 and Bax at mRNA and protein level was increased after UV exposure (P ＜ 0.01).But for Bcl-2 the expression both at mRNA and protein level was decreased after a 1 day-exposure(P ＜0.01) and was increased after UV exposure of 5 days and 7 days(P ＜ 0.01).The mRNA expression level of p53 was positively correlated with the Bax (r =0.952,P ＜ 0.01).The expression of p53 and Bax protein were increased after UV exposure,and transferred from cytoplasm to nucleus;While the expression of Bc1-2 protein was decreased at 1 day after UV exposure,and increased gradually at 3 days,5 days and 7 days.Conclusion The apoptotic process of rat lens epithelial cells induced by ultraviolet may be related with p53 regulating the expression of Bax/Bcl-2.
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The cultivation of general practitioner is a key to improve China's community-based medical and health care system.Zunyi Medical University,in the process of cultivating tuition-free students with the mode of serving and staying in the countryside after their graduation,always upholds the principles of being oriented by post competence,being led by medical humanities,optimizing featured courses about general practitioner,carrying out students-centered teaching methodology,focusing on training students' autonomous learning ability,and fully building the practical teaching system,so as to strive to cultivate high-quality talents in the field of general medicine to make them better serve the rural community-based medical institutions.
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Background Various studies demonstrated that the apoptosis of lens epithelial cells(LECs) is associated with the overexpression of the c-myc gene in LECs induced by galactose.Inhibiting the abnormal expression of the c-myc gene in LECs is an effective approach to mitigate the pathogenesis and development of cataract.Objective The goal of this study is to investigate the inhibitory effects of c-myc antisense oligodeoxynucleotide(c-myc ASODN) on the apoptosis of LECs in the eye with galactose-induced cataract.Methods Galactose-induced cataract models were established by the retrobulbar injection of 0.2 mL of 20% galactose once per day.Lipo-antisense oligodeoxynucleotide(Lipo-ASODN,0.2 mL) was retrobulbarly injected 4 hours after the injection of galactose at one-day intervals.The animals were sacrificed and lenses were obtained to evaluate the apoptosis of LECs and the effect of c-myc ASODN on LECs apoptosis induced by galactose was examined by TUNEL assay after 7,14 and 24 days.The ultrastructural changes of LECs were examined under the transmission electron microscopy(TEM).Results A significant difference in the apoptotic rate of LECs was found among the 7 day,14 day and 24 day groups(F_(7 days)=3 418.495,P＜0.01;F_(14 days)=1137.555,P＜0.01;F_(24 days)=2198.871,P＜0.01).The apoptotic rate of LECs in the galactose group was markedly higher than that in the normal saline solution group 7 days,14 days and 24 days after the experiment(P＜0.01).The apoptotic rate of LECs in the galactose+lipo+ ASODN group significantly declined in comparison to the galactose group after 7 days,14 days and 24 days(P＜0.05).TUNEL assay showed the condensation,breakage and irregularity of the nuclei of apoptotic cells in the galactose group.The destruction of the ultrastructure of the cells and organelles were observed under the transmission electron microscope.Conclusion Galactose induces apoptosis of LECs in cataractogenesis.C-myc ASODN inhibits apoptosis of LECs induced by galactose.
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Objective To explore the relationship between galactose-induced apoptosis of lens epithelial cells and cataractogenesis.Methods Galactose was injected retrobulbarly into Wistar rats.The opacity of lens was examined by slit lamp.The expression of C-MYC was measured by flow cytometer.The apoptosis of lens epithelial cells was observed by TUNEL techniques.Results More than 50 percent of lenses were in the Ⅱ stage of cataractogenesis on the(7 d) after galactose induction;Eighty-seven percent of lenses were in the Ⅳ-Ⅴ stage on the(14 d),and 100 percent of lenses were in the Ⅳ-Ⅴ stage on the(24 d).The rate of apoptotic cell on the(7 d) and on the(14 d) after galactose induction was respectively 5.6%～8.4% and 30.2%～41.8%,and 60% apoptotic cells were observed on the(24 d).The levels of C-MYC expression in lens epithelial cells induced by galactose on the(7 d) and(14 d) were higher than that of normal lenses,but the level of C-MYC expression backed to normal level on the(24 d).Conclusion It was suggested that there are some apoptosis of the lens epithelial cells during galactose-induced cataractogenesis and that the apoptosis of the lens epithelial cells might be caused by higher expression of C-MYC.
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Objective To investigate the effects of antisense oligodeoxynucleotide c-myc (c-myc ASODN) on the growth of rabbit lens epithelial cells (LECs) induced by galactose. Methods Based on the translation initiation region of the second extron of c-myc gene, oligonucleotides were synthesized and modified with phosphorothioate. Lens epithelial cells were treated with galactose and c-myc ASODN. The effects of c-myc ASODN on the growth of LECs, cultured in high concentration of galactose, were observed by cytometry and MTT. Results The results demonstrated that c-myc ASODN at the doses of 2.5-10.0 ?mol/L inhibited the growth of LECs induced by galactose in a dose-dependent manner. Conclusion These results indicate that c-myc ASODN can inhibit the growth of LEC induced by galactose.
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Objective Using the soaking seedling method to introduce wild Gastrodia elata DNA into potato plantlets and analyze gastrodin of transformed Solanum tuberosum.Methods After the tuber grown up,the solution of gastrodin was extracted from the potatoes which were transformed by wild G.(elata) DNA.The transformed plants were scaned by ultraviolet.PCR was used to analyze GAFP gene by SDS-PAGE.TLC was used to analyze the gastrodin of transformed S.tuberosum.Results(1)The 21 transformed plants had obvious absorption peak at 220 nm in 200 transformed S.tuberosunr.(2)The fragment of wild G.elata GAFP gene was found by PCR from 5 transformed plants.(3)The protein expressions in transgenic S.tuberosum were obviously different from the normal S.tuberosum;a band was detected in transgenic S.tuberosum and wild G.elata,but wasn't detected in normal S.tuberosum.(4)The expression of the gastrodin was detected by TLC from three transformed plants. Conclusion It is feasible to use the soaking seedling method to introduce exogenous DNA into plants.