Effects of ginkgolide B on the development of embryonic rat midbrain neurons

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Ἱππολύτη Βιτάλης
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1 Journal of Jiangsu UniversityMedicine Edition Vol. 21 No. 4 Jul B B ginkgolide B GB kainic acid KA NGF B NGF + KA GB + KA MTT 72 h B P < P < d P < B B P < B NGF P < P < B B B R971 A Effects of ginkgolide B on the development of embryonic rat midbrain neurons TAN Cai-hong 1 MIAO Li-yan 2 1. Department of Pharmacy the Affiliated Hospital of Jiangsu University Zhenjiang Jiangsu Department of Pharmacy the First Affiliated Hospital of Soochow University Suzhou Jiangsu China AbstractObjectiveTo study the effects of ginkgolide B on the normal neurons of embryonic rat midbrain and on the embryonic rat midbrain neurons injured by kainic acidka. MethodsThe experimental objects were divided into control positive control NGF ginkgolide BGB damage groupka NGF + KA GB + KA groups. The survival ratio and the colony differentiation ratio of embryonic rat midbrain neurons were estimated by MTT assay and microscopy respectively. Results1Survival ratio of midbrain nerve cellscompared with the control group the survival ratio of embryonic rat midbrain neurons in ginkgolide B groups were increasing significantly when it was cultured for 72 hoursp < P < The cell survival ratio in kainic acid group was lower obviously than that of control group at days of threep < The nerve cells exposed to kainic acid showed characteristic change of injury which could be relieved by the treatment of ginkgolide B and ginkgolide B showed the dose-dependent mannerp < Colony differentiation ratio of embryonic rat midbrain neuronscompared with the control group the colony differentiation ratio of ginkgolide B and NGF groups were increasing obviouslyp < and KA can inhibit the colony differentiation ratio of embryonic rat midbrain neurons sharplyp < Ginkgolide B can partly prevent the embryonic rat midbrain cells from damage by kainic acid. ConclusionGinkgolide B can promote the development of embryonic rat midbrain neurons and prevent the nerve cells from damage by kainic acid. Key wordsginkgolide Bnerve growth factorkainic acidembryonic rat midbrain nerve cell 1966

2 Wistar 75% 5 min PBS 0. 25% 37 5 min 1 - NGF % Bginkgolide B GB /ml 10 mg /L CO % CO 2 24 h 1 2 ~ 3 MTT B B NSE 1. 1 Wistar SP NSE DAB Wistar 250 g d % DMEM 10 d NSE B MTT NSE 95% kainate KA Sigma 1. 6 MTT DMEM B Gibco /ml μl 24 h B 4 6 B mg /L neuron specific enolase NSE72 h 4 h 5 g /L MTT SP DAB 20 μl 4 h DMSO150 μl min 490 nm = / Gilson Leica Mz12. 5 / B Leica CO 2 Shellab 8 1 Olympus TDL-5C 2 NGF50 mg /L 3 B 50 mg /L μmol /

3 4. B 283 L 5 NGF 50 mg /L+ KA100 μmol /L6 B mg /L+ KA100 μmol /L 1 B % CO 2 24 n = 6 x ± s h Tab 1 Effects of ginkgolide B in different concentration on the survival ratio of embryonic rat midbrain 1. 7 B neurons cultured in different groups B 3-4 D mg /L % ± /L 20 μlco 2 4 h ± a d ± b % 1 ~ 2 h PBS ± a Giemsa ± b ap < bp < x ± s 100% 2. 2 B 1. 8 x ± s SPSS P < SNK 96 h 32. 8% q α B B NGF + KA 72 h B ~ 100 mg /L 96 h NGF50 mg /L P < P < B ~ 100 mg /L 123% 2 72 h P < GB + KA 72 h P < B 125% B50 mg /L Tab 2 2 MTT n = 6 x ± s Absorbance value detected by MTT of embryonic rat midbrain neurons cultured in different groups 24 h 48 h 72 h 96 h ± ± ± ± NGF ± ± ± a b B ± ± ± a b ± a b ± a b ± ± ± a ± a NGF + KA ± ± ± b ± b GB25 mg /L+ KA ± ± ± b ± b GB50 mg /L + KA ± ± ± b ± b GB100 mg /L+ KA ± ± ± b ± b F P ap < bp < 0. 05

284 21 2. 3 P < 0. 01 B 4 h NGF + KA 3 1 10 h 3 24 h n = 6 x ± s Tab 3 Colony differentiation ratio of embryonic rat midbrain neurons cultured in different groups 10 d B NGF NGF 94. 3 ± 3. 20 125.

4 P < B 4 h NGF + KA h 3 24 h n = 6 x ± s Tab 3 Colony differentiation ratio of embryonic rat midbrain neurons cultured in different groups 10 d B NGF NGF ± ± a % B ± GB + KA 134 B NGF NGF + KA GB50 mg /L+ KA P ± a ± b ± b < GB + KA ap < bp < Fig 1 1 Giemsa 200 Colony differentiation shape of embryonic rat midbrain neurons cultured in diferent groups mg /L 50 mg /L B ginkgo biloba extract EGB 24% 6% 5 A B C M J B B B 25 ~ 100 mg /L d GB + KA B B B NGF B 8-10 B NGF BDNF -3 NT B h B ~ 288

5 Kohler GMilstein F. Continuous cultures of fused cells secreting antibody of predefined specificity J. Nature J Pahwa RJaggaiahgari SPahwa Set al. Isolation and 3O'Neill JPMachanoff RHsie AW. Phenotypic expression time of mutagen-induced 6-thioguanine resistance in therapy J. J Immunol Methods Chinese hamster ovary cells CHO /HGPRT system expansion of human natural T regulatory cells for cellular 79 expression in division-arrested cell cultures J. Environ Mutagen Sugawa HMori TImura H. Establishment of 8-azaguanine-resistant mutants from rat thyroid cell line FRTL-5 J. Mol Cell Endocrinol Littlefild JW. The use of drug-resistant markers to study the hybridization of mouse fibroblasts J. Exp Cell Res Schmitt TMZúiga-Pflücker JC. Induction of T cell development from hematopoietic progenitor cells by deltalike-1 in vitro J. Immunity Sharma RSung SSJu CYet al. Regulatory T-Cell Treghybridoma as a novel tool to study Foxp3 regulation and Treg fate J. J Autoimmun2011 May 27. Epub ahead of print J Ding Y Zeng YS Zhang W et al. The effects of ginkgolide B of various consistency on the differentiation of neu- 2. J ral stem cells J. Acta Anatomica Sinica NGF J. from leaves of Ginkgo biloba on glutamate neurotoxicity J C E 11Kusmic C Basta G Lazzcrini G et al. The effect of Ginkgo biloba in isolated ischemic /reperfused rat heart J a link between vitamin E preservation and prostaglandin biosynthesis J. J Cardiovasc Pharmacol B J bcl-2 bax J. J B Ca 2 + J Antagonistic effects of extract J J

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