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摘要:
目的 探讨中药复方筋脉通对糖尿病大鼠背根神经节尼氏染色及NADPH氧化酶、诱导型一氧化氮合酶(inducible nitric oxide synthase, iNOS)表达的影响。 方法 70只雄性Sprague Dawley大鼠腹腔注射链脲佐菌素诱导糖尿病大鼠模型后, 随机分为模型组、筋脉通大、中、小剂量组和维生素C对照组, 每组14只, 同时设正常对照组(n=10);筋脉通大、中、小剂量组分别按成人剂量20倍、10倍、5倍、维生素C组按成人剂量10倍灌胃给药, 模型组和正常对照组予等量蒸馏水灌胃; 16周后对各组背根神经节切片行尼氏染色, 免疫组化法检测NADPH氧化酶p22 phox亚基和iNOS的表达。 结果 模型组神经元胞体中尼氏体缺失, 筋脉通大、中剂量组尼氏体改变明显轻于模型组; 筋脉通各剂量组NADPH氧化酶p22 phox亚基、iNOS表达均明显低于模型组(P < 0.05)。 结论 糖尿病大鼠周围感觉神经节存在氧化应激反应, 筋脉通可减轻氧化应激损伤, 保护神经元。 Abstract:Objective To investigate the influence of Chinese compound formula Jinmaitong on the Nissl's staining of dorsal root ganglion (DRG) of diabetic rats and on the expressions of NADPH oxidase and inducible nitric oxide synthase (iNOS). Methods Dabetic rat models were induced after the intraperitoneal injection of streptozotocin (STZ) to 70 male Sprague Dawley rats. These rats were then divided into five groups (n=14):model group (placebo), Jinmaitong high-dose group (20-fold dose recommended for human), Jinmaitong middle-dose group (10-fold dose recommended for human), Jinmaitong low-dose group (5-fold dose recommened for human), and vitamin C group (10-fold dose recommened for human). Meanwhile, 10 rats without STZ intervention were set as normal group. Drugs (or placebo) were intragastically administered once per day. Sixteen weeks later, DRG tissues were isolated for Nissl's staining as well as immunohistochemistry staining to detect the expressions of NADPH oxidase p22phox and iNOS. Results Nissl's bodies depleted obviously in model group. In Jinmaitong high-dose and middle-dose groups, the morphologic abnormality of Nissl's bodies was slight. Compared to model group, the expression of NADPH p22 phox and iNOS in all Jinmaitong groups reduced significantly (P < 0.05). Conclusions The study gives evidence that oxidative stress exists in peripheral sensory nerve ganglion of diabetic rats. Jinmaitong can counteract oxidative stress and protect neurons. -
图 1 尼氏体在正常对照组(A)、STZ诱导的糖尿病模型组(B)、维生素C组(C)及筋脉通小剂量组(D)、中剂量组(E)、大剂量组(F)大鼠DRG组织中的形态学变化(尼氏染色，× 400)
STZ:链脲佐菌素; DRG:背根神经节
图 2 NADPH氧化酶p22phox亚基在正常对照组(A)、STZ诱导的糖尿病模型组(B)、维生素C组(C)及筋脉通小剂量组(D)、中剂量组(E)、大剂量组(F)大鼠DRG组织中的表达结果(免疫组化染色，× 200)
STZ、DRG:同图 1
图 3 诱导型一氧化氮合酶在正常对照组(A)、STZ诱导的糖尿病模型组(B)、维生素C组(C)及筋脉通小剂量组(D)、中剂量组(E)、大剂量组(F)大鼠DRG组织中的表达结果(免疫组化染色，× 200)
STZ、DRG:同图 1
表 1 各组大鼠血糖变化(mmol /L，x±s)
组别 n 干预前 用药后 4周 8周 12周 16周 正常对照组 10 5. 0 ± 0. 7 5. 1 ± 0. 3 5. 4 ± 0. 8 6. 1 ± 0. 1 5. 4 ± 0. 4 糖尿病模型组 11 21. 1 ± 3. 1* 21. 1 ± 1. 8* 23. 2 ± 1. 0* 23. 9 ± 2. 4* 21. 9 ± 0. 7* 维生素C组 12 21. 3 ± 3. 1* 20. 7 ± 1. 4* 23. 8 ± 0. 5* 23. 8 ± 0. 5* 22. 9 ± 1. 1* 筋脉通 小剂量组 12 20. 8 ± 3. 2* 21. 2 ± 1. 9* 22. 9 ± 0. 9* 22. 8 ± 1. 6* 22. 3 ± 0. 7* 中剂量组 13 21. 0 ± 3. 1* 22. 8 ± 1. 7* 23. 5 ± 0. 8* 23. 3 ± 2. 5* 21. 2 ± 1. 8* 大剂量组 12 20. 8 ± 3. 3* 22. 4 ± 2. 1* 21. 9 ± 0. 7* 23. 4 ± 1. 1* 22. 1 ± 0. 7* 与正常对照组比较，* P＜0. 01 
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表 2 各组大鼠体重变化(g，x±s)
组别 n 造模后 用药后 4周 8周 12周 16周 正常对照组 10 257. 2 ± 20. 2 532. 5 ± 10. 5 578. 3 ± 24. 4 604. 1 ± 33. 1 641. 2 ± 15. 8 糖尿病模型组 11 256. 4 ± 17. 3 303. 6 ± 10. 1* 321. 5 ± 9. 3* 431. 1 ± 6. 8* 468. 1 ± 7. 2* 维生素C组 12 257. 1 ± 33. 8 301. 2 ± 9. 6* 329. 7 ± 8. 3* 449. 2 ± 10. 7* 477. 2 ± 8. 9* 筋脉通 小剂量组 12 258. 1 ± 22. 1 284. 9 ± 10. 1* 326. 5 ± 10. 4* 426. 2 ± 8. 5* 459. 5 ± 10. 7* 中剂量组 13 256. 2 ± 18. 8 303. 3 ± 10. 7* 324. 5 ± 10. 9* 452. 6 ± 9. 3* 480. 6 ± 12. 3* 大剂量组 12 257. 9 ± 23. 9 299. 9 ± 16. 6* 341. 7 ± 14. 3* 442. 2 ± 13. 6* 467. 8 ± 14. 8* 与正常对照组比较，* P＜0. 01
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表 3 各组大鼠干预16周后机械痛阈值(g，x±s)
组别 n 机械痛阈值 正常对照组 10 78. 69 ± 8. 13 糖尿病模型组 11 35. 32 ± 12. 06* 维生素C组 12 59. 49 ± 3. 42*† 筋脉通 小剂量组 12 52. 87 ± 6. 46*† 中剂量组 13 74. 69 ± 9. 26†‡ 大剂量组 12 61. 71 ± 1. 95* † 与正常对照组比较，* P＜0. 01;与糖尿病模型组比较，†P＜0. 01;与维生素C组比较，‡P＜0. 01
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表 4 各组大鼠DRG组织中NADPH氧化酶p22phox亚基和iNOS蛋白表达(n = 5，x±s)
组别 p22phox亚基表达IOD值 iNOS表达IOD值 正常对照组 6. 17 ± 1. 86 12. 51 ± 1. 33 糖尿病模型组 64. 57 ± 1. 58* 71. 91 ± 2. 74* 维生素C组 50. 09 ± 6. 19* † 52. 88 ± 5. 64* †† 筋脉通 小剂量组 31. 24 ± 2. 96* ††‡ 49. 24 ± 2. 61* †† 中剂量组 27. 09 ± 3. 46*††‡‡ 35. 86 ± 2. 00*††‡ 大剂量组 10. 48 ± 2. 70††‡‡ 24. 68 ± 1. 77*††‡‡ DRG:背根神经节; iNOS:诱导型一氧化氮合酶; IOD:积分吸光度; 与正常对照组比较，* P＜0. 01;与糖尿病模型组比较，†P＜0. 05，†† P＜0. 01;与维生素C组比较，‡P＜0. 05，‡‡P＜0. 01与正常对照组比较，* P＜0. 01;与糖尿病模型组比较，†P＜0. 01;与维生素C组比较，‡P＜0. 01
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