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

21 4 2011 7 Journal of Jiangsu UniversityMedicine Edition Vol. 21 No. 4 Jul. 2011 B 1 2 1. 2120012. 215006 B ginkgolide B GB kainic acid KA NGF B NGF + KA GB + KA MTT 72 h B P < 0. 05 P < 0. 01 3 d P < 0. 05 B B P < 0. 05 B NGF P < 0. 01 P < 0. 01 B B B R971 A 1671-7783201104 - 0281-04 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 2120012. Department of Pharmacy the First Affiliated Hospital of Soochow University Suzhou Jiangsu 215006 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 < 0. 05 P < 0. 01. The cell survival ratio in kainic acid group was lower obviously than that of control group at days of threep < 0. 05. 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 < 0. 05. 2Colony differentiation ratio of embryonic rat midbrain neuronscompared with the control group the colony differentiation ratio of ginkgolide B and NGF groups were increasing obviouslyp < 0. 01 and KA can inhibit the colony differentiation ratio of embryonic rat midbrain neurons sharplyp < 0. 01. 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

282 21 1. 4 3-4 14 Wistar 75% 5 min PBS 0. 25% 37 5 min 1 - NGF 2 200 92% Bginkgolide B GB 4 10 6 /ml 10 mg /L CO 2 37 5% CO 2 24 h 1 2 ~ 3 MTT B B 1. 5 1 NSE 1. 1 Wistar SP 1 100 NSE DAB Wistar 250 g 2 1 600 0 d14 1. 2 10% DMEM 10 d NSE B MTT NSE 95% kainate KA Sigma 1. 6 MTT DMEM 1 250 1. 6. 1 B Gibco 4 10 6 /ml 96 50 μl 24 h B 4 6 B 12. 5 25 50 100 mg /L neuron specific enolase NSE72 h 4 h 5 g /L MTT SP DAB 20 μl 4 h DMSO150 μl 1. 3 15 min 490 nm = / Gilson Leica Mz12. 5 / 1. 6. 2 B Leica CO 2 Shellab 8 1 Olympus TDL-5C 2 NGF50 mg /L 3 B 50 mg /L 4 100 μmol /

4. B 283 L 5 NGF 50 mg /L+ KA100 μmol /L6 B25 50 100 mg /L+ KA100 μmol /L 1 B 96 37 5% CO 2 24 n = 6 x ± s 48 72 96 h 1. 6. 1 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 % 96 4 0. 584 ± 0. 026 100 10 6 /L 20 μlco 2 4 h 12. 5 0. 615 ± 0. 022 a 105. 3 1. 6. 2 6 10 d 25 0. 633 ± 0. 025 b 108. 4 4% 1 ~ 2 h PBS 50 0. 668 ± 0. 056 a 114. 3 2 Giemsa 100 0. 684 ± 0. 048 b 117. 1 ap < 0. 05 bp < 0. 01 50 x ± s 100% 2. 2 B 1. 8 x ± s SPSS 10. 0 P < 0. 05 SNK 96 h 32. 8% q α 0. 05 2 2. 1 B B NGF + KA 72 h B 12. 5 ~ 100 mg /L 96 h NGF50 mg /L P < 0. 05 P < 0. 01 1 B 12. 5 ~ 100 mg /L 123% 2 72 h P < 0. 05 GB + KA 72 h P < 0. 05 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 0. 365 ± 0. 04 0. 371 ± 0. 02 0. 583 ± 0. 04 0. 685 ± 0. 06 NGF 0. 372 ± 0. 03 0. 396 ± 0. 04 0. 656 ± 0. 04 a b B 0. 362 ± 0. 03 0. 391 ± 0. 02 0. 662 ± 0. 06 a b 0. 856 ± 0. 05 a b 0. 844 ± 0. 05 a b 0. 345 ± 0. 02 0. 342 ± 0. 02 0. 242 ± 0. 03 a 0. 225 ± 0. 02 a NGF + KA 0. 354 ± 0. 02 0. 369 ± 0. 05 0. 532 ± 0. 06 b 0. 575 ± 0. 07 b GB25 mg /L+ KA 0. 346 ± 0. 04 0. 363 ± 0. 05 0. 394 ± 0. 06 b 0. 426 ± 0. 04 b GB50 mg /L + KA 0. 352 ± 0. 04 0. 364 ± 0. 04 0. 527 ± 0. 05 b 0. 568 ± 0. 04 b GB100 mg /L+ KA 0. 358 ± 0. 03 0. 374 ± 0. 04 0. 539 ± 0. 05 b 0. 605 ± 0. 06 b F 0. 449 1. 062 39. 254 83. 613 P 0. 863 0. 410 0. 000 0. 000 ap < 0. 05 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. 9 ± 5. 92 a % 100 133 B 126. 6 ± 7. 20 GB + KA 134 B NGF NGF + KA GB50 mg /L+ KA P 27. 7 ± 4. 92 a 81. 6 ± 5. 95 b 77. 4 ± 6. 00 b 29. 4 86. 5 82. 1 < 0. 01 GB + KA ap < 0. 01 bp < 0. 01 Fig 1 1 Giemsa 200 Colony differentiation shape of embryonic rat midbrain neurons cultured in diferent groups 3 100 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 5-7 10 d GB + KA B B B NGF B 8-10 B NGF BDNF -3 NT-3 11-13 B 1. 72 h B 12. 5 ~ 288

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