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Background Minocycline possesses neuroprotective effect in a variety of animal models and clinical trials of central nervous system,but whether it works on optic nerve injury remains unclear.Objective This study aimed to observe the protective effects of minocycline on retinal ganglion cells (RGCs) in the early stage of optic nerve crush and explore its mechanism.Methods One hundred and thirty-six clean C57BL/6J mice were randomly divided into normal control group,normal saline solution group and minocycline group.The optic nerve crush injury models were induced in the left eyes of the mice in the normal saline solution group and minocycline group by a cross-action forceps for 3 seconds.Minocycline was injected intraperitoneally in the minocycline group firstly 45 mg/kg(0.4 ml) and followed by 22.5 mg/kg per day after 24 hours until sacrifice of the animals,and the equivalent volume of normal saline solution was injected in the same way in the normal saline solution group.The mice were euthanized at 4,7,11,14 days postoperatively and the left eyeballs were collected.Retinal flat mounts and DAPI staining was used to observe and compare the change of RGCs density among different groups and various time points.Apoptosis of mice RGCs were assessed by TdT-mediated dUTP nick end labeling (TUNEL).Real-time polymerase chain reaction (real-time PCR) was used to detect the expression of CD11b mRNA in retinal microglials.Results DAPI staining in retinal flat mounts showed that the average RGCs density was (77.50±2.38)/0.01 mm2 and (70.00±2.94) /0.01 mm2 in the 4th and 7th day after modeling in the normal saline solution group,and those in the minocycline group were (88.75 ± 2.36) /0.01 mm2 and (81.00 ± 3.92)/0.01 mm2,with significant differences between the two groups (t4d =-6.708,P＜0.01 ;t7d =--4.491,P＜0.01).The apoptotic RGCs were (12±1)/mm and (4±1)/mm in the normal saline solution,which were significantly more than (4±1)/mm and (1±0)/mm in the minocycline group (t4 d =12.832,P＜0.01 ; t7d =3.455,P =0.026).However,no significant difference was found in apoptotic RGCs in postoperative 11 days and 14 days between the normal saline solution group and the minocycline group (P =0.708,0.777).The expressing levels of CD11 b mRNA in the retinal microglials were significantly higher in the 4th and 7th day in the normal saline solution group than those in the minocycline group (t4 d =8.312,P＜0.01 ;t7d=5.407,P＜0.01),but were not significantly different in the 11st and 14th day after modeling between the two groups (P=0.055,0.170).Conclusions Minocycline can play a neuroprotective effect on RGCs in the early stage of optic nerve crush in mice by inhibiting microglia activation and decreasing RGCs apoptosis.
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<p><b>BACKGROUND</b>Currently, no medicine is available that can prevent or treat neural damage associated with optic nerve injury. Minocycline is recently reported to have a neuroprotective function. The aims of this study were to exarmine the neuroprotective effect of minocycline on retinal ganglion cells (RGCs) and determine its underlying mechanisms, using a mouse model of optic nerve crush (ONC).</p><p><b>METHODS</b>ONC was performed in the left eye of adult male mice, and the mice were randomly divided into minocycline-treated group and saline-treated control group. The mice without receiving ONC injury were used as positive controls. RGC densities were assessed in retinal whole mounts with immunofluorescence labeling of βIII-tubulin. Transmission electron microscopy was used to detect RGC morphologies, and Western blotting and real-time PCR were applied to investigate the expression of autophagy markers LC3-I, LC3-II, and transcriptional factors nuclear factor-κB1 (NF-κB1), NF-κB2.</p><p><b>RESULTS</b>In the early stage after ONC (at Days 4 and 7), the density of RGCs in the minocycline-treated group was higher than that of the saline-treated group. Electron micrographs showed that minocycline prevented nuclei and mitochondria injuries at Day 4. Western blotting analysis demonstrated that the conversion of LC3-I to LC3-II was reduced in the minocycline-treated group at Days 4 and 7, which meant autophagy process was inhibited by minocycline. In addition, the gene expression of NF-κB2 was upregulated by minocycline at Day 4.</p><p><b>CONCLUSION</b>The neuroprotective effect of minocycline is generated in the early stage after ONC in mice, partly through delaying autophagy process and regulating NF-κB2 pathway.</p>
Subject(s)
Animals , Male , Mice , Autophagy , Minocycline , Therapeutic Uses , NF-kappa B p52 Subunit , Metabolism , Optic Nerve Injuries , Drug Therapy , Metabolism , Retinal Ganglion Cells , MetabolismABSTRACT
Objective To explore the change of the total level of lgE and specific allergen for acute exacerbation of bronchial asthma and to offer evidence for the prevention and cure of acute exacerbation of bronchial asthma.Methods The usual inhalant and ingested antigens were chosen in asthma patients,and the allergens were detected by ELISA in sera of acute exacerbation of bronchial asthma patients.Results Total positive rate of lgE was 92％ (48/52),dust mites 61.2％ (25/52),house dust 17％ (9/52),shrimp and crab 15％ (8/52),mould 9％ (5/52).Conclusion The main allergen of acute asthma exacerbation is dust mites.House dust,shrimp and crab,and mould folloo.The rats of pollen or weed are very low.
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Objective To evaluate the immunogenic stability and hereditary stability of Neisseria meningitides serogroup W135/Y[CMCC(B)29037/CMCC(B)29028]within all the passages,which isolated from china.Methods The toxicity of the 3rd,5th,10th,15th,20th,25th and 30th passage of the Neisseria meningitidis was assayed in mice.Serological detection and biochemical detection were measured,and immunized mice subcutaneously.The antigeeicity of each passage of Neisseria meningitides serogroup W135/Y were measured by serum bactericidal test and the indirect ELISA.With the 30 passage of Neisseria meningitides serogroup W135/Y,the effect to the encephalic tissue was measured in mice.Fermented the Neisseria meningitides serogroup W135/Y with 30 passage and purified the capsular polysaccharide,then analyzed the quality respectively.Results The LD50 of the strains CMCC(B)29037/29028 of each passage was low(LD50 ≥ 109),and all the 30logical detection and all the 30 passage of the two strains were half in the tube agglutination.Glucose and maltose fermentation test were positive.Fructose,sucrose and lactose fermentation test were negative.The GMT of immunogenicity were 1114 and 2229 respectively and all the 30 passage were more than 640 and 1040 respectively.After Immunization with individual 30 passage of the Neisseria meningitides,the titer in serum bactericidal assay(SBA)and indirect ELISA were no difference.The capsular polysaccharide purified from Neisseria meningitides serogroup W135/Y met the quality standard.Conclusion Neisseria meningitides serogroup W135/Y,CMCC(B)29037/29028,used in the manufacture of the meningococcal conjugate vaccine,are stable in the toxicity,antigenicity,immunogenicity.Serological detection and biochemical detection are qulified,and the capsular polysaccharide has met the quality standard.
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Objective: To investigate the effects of different concentrations of vitamin E (VE) on angiogenesis and its possible mechanism. Methods: The rings of rat aorta were embedded in gels of collagen and cultured in a serum free medium for 28 days. Curves of microvessels growth were generated by counting the number of newly formed microvessels every day with an inverted microscope. Photos were taken at the same time. Expression of factor FVIII related antigen (FVIII RAg) in newly formed microvessels was detected by immunohistochemistry. Concentration change of FVIII R in medium was evaluated by ELISA. Results: Both 0.2 g/L and 0.1 g/L VE showed significant inhibitory effect on angiogenesis (P0.05). The inhibitory effect of VE was expressed in the phases of microvessel growth and decline (10-25 d). Conclusion: 1.Three dimensional collagen gels culture of microvessel in serum free medium can be used as a sensitive assay for study of soluble and solid phase angiogenic agonists and antagonists. 2. Both 0.1 g/L and 0.2 g/L VE inhibit angiogenesis, and VE has no toxicity and no drug resistance.