清燥布津法对干燥综合征小鼠下颌腺AQP_5表达的影响
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摘要
目的:建立干燥综合征(Sj(?)gren's syndrome SS)动物模型,观察清燥布津法对SS小鼠模型下颌腺aquaporin-5(AQP_5)表达的影响。方法:雌性BALB/c小鼠颌下腺匀浆离心后的上清液加弗氏完全佐剂(CFA)制成乳化免疫抗原,采用多点注射法免疫同系鼠,同时注射白百破疫苗建立SS小鼠模型。以环戊硫酮为对照组,观察中药对SS小鼠模型唾液分泌、病理及下颌腺AQP_5(免疫组化法)表达的影响。结果:模型组与正常对照组比较,小鼠的体重、唾液流率下降,饮水量增加。下颌腺组织中淋巴细胞重度浸润。腺体破坏。免疫组化方法显示:AQP_5在腺体的分布和表达减少或消失。中药高剂量组与模型组比较,小鼠的体重、唾液流率增加,饮水量减少。下颌腺组织中只有淋巴细胞的轻度浸润。腺体的结构正常,无破坏。AQP_5在腺泡、闰管、导管等处均有较强表达。中药高剂量组疗效优于其他治疗组。结论:清燥布津法有明显抑制腺体破坏,改善唾液分泌功能的作用,其作用机理可能与抑制免疫炎症,促进AQP_5在小鼠下颌腺的表达有关。
Objective: To establish a experimental SjOgren's syndrome (SS) model and observe the effect of QZBJ therapeutic method on expression of aquaporin-5 (AQP5) in SS mice. Methods: BALB/c mice were injected with homologous antigen of submandibular gland tissues plus complete Freund's adjuvant ( CFA ) plus diptheria-tetanus toxoids-pertussis to enhance the immunological reaction. The same immunological way repeated twice to duplicate SS model. Treating with Anethol Trithione as control group, saliva flow rate test, histopathological findings and AQP5 expression in submandibular glands were observed. Results: Obvious losing of weight and saliva, increasing weight of drinking water were found in SS models, and there were also massive lymphocytic infiltration and reduced number of the acinar in submandibular gland of BALB/c nice, where AQP5 expression decreased or disappeared. High-dosage QZBJ Mixtrue could obviously increase weight and excretion of salivary glands while decrease drinking water. AQP5 was strongly expressed in the acinar and the secretory ducts in high-dose QZBJ Mixtrue group. Conclusion: QZBJ therapeutic method obviously inhibits lesions of salivary glands and improves functions of salivary glands. QZBJ therapeutic method might treat SS by inhibiting immunlogical inflammation and affecting expression of AQP5 in salivary glands.
Objective: To establish a experimental SjOgren's syndrome (SS) model and observe the effect of QZBJ therapeutic method on expression of aquaporin-5 (AQP5) in SS mice. Methods: BALB/c mice were injected with homologous antigen of submandibular gland tissues plus complete Freund's adjuvant ( CFA ) plus diptheria-tetanus toxoids-pertussis to enhance the immunological reaction. The same immunological way repeated twice to duplicate SS model. Treating with Anethol Trithione as control group, saliva flow rate test, histopathological findings and AQP5 expression in submandibular glands were observed. Results: Obvious losing of weight and saliva, increasing weight of drinking water were found in SS models, and there were also massive lymphocytic infiltration and reduced number of the acinar in submandibular gland of BALB/c nice, where AQP5 expression decreased or disappeared. High-dosage QZBJ Mixtrue could obviously increase weight and excretion of salivary glands while decrease drinking water. AQP5 was strongly expressed in the acinar and the secretory ducts in high-dose QZBJ Mixtrue group. Conclusion: QZBJ therapeutic method obviously inhibits lesions of salivary glands and improves functions of salivary glands. QZBJ therapeutic method might treat SS by inhibiting immunlogical inflammation and affecting expression of AQP5 in salivary glands.
引文
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[19] King LS, Agre P. Pathophysiology of the aquaporin water channels. Ann Rev Physiol. 1996, 58: 619～648
[20] Cheng A, Hock AN, Yeager M, et al. Three-dimensional organization of a human water channel. Nature, 1997, 387: 627～630
[21] Lih Lees, Jap Bk. Molecular design of aquaporin-1 water channel as revealed by electron crystallography. Nat Strut Biol. 1997, 4: 263～265
[22] Ma T, Song Y, Gillespie A, et al. Defective secretion of saliva in transgenic mice lacking aquaporin-5 water channels[J]. J Biol chem, 1999, 274 (29): 20 071-20 074
[23] Takagi K, Yamaguchi K, Sakurai T, et al. Secretion of saliva in X-irradiated rat submandibular glands[J]. Radiat Res, 2003, 159 (3): 351-360
[24] Steinfeld S, Cogan E, king LS, et al. Abnormal distribution of aquaporin-5 water channel protein in salivary glands from SjOgren' syndrome patients Lab invest, 2001, 81: 143-148
[25] Ruddy S, Harris ED Jr, Sledye CB. Kelley' textbook of rheumatology [M]. 6th ed. Philadelphia: W. B. Saunders Company, 2001: 1028
[26] Rose NR, Mackay IR. The autoimmune diseases[M]. 3 rd ed. San Diego: Academic Press, 1998: 381
[27] CuelloC, Pallladinetti P, Tedla N, et al. Chemokine expression and leucocyte infiltration SjOgren' syndrome[J]. Br J Rheumatol, 1998, 37: 779
[28] Anaya JM, Talal N. SjOgren's syndrome Comes of age[J]. Semin Arthritis Rheum, 1999, 28: 355
[29] Nakamura H, Kawakami A, Tominaga M, et al. Expression of CD40/CD40 ligand and Bcl-2 family proteins in labial salivary glands of patients with SjOgren's syndrome[J]. Lab Invest, 1999, 79: 261
[30] Saegusa K Ishimaru N, Yanagi K, et al. Treatment with anti-CD86 costimulatory molecule prevents the autoimmune lesions in murine SjOgren' syndrome through up-regulated Th2 rsponse[J]. Clin Exp Immunol, 2000, 119: 354
[31] Stott DI, Hiepe F, Hummel M, et al. Antigen driven clonal proliferation of B cells with in the target tissue of an autoimmune disease: the salivary glands of patients with SjOgren' syndrome[J]. J Clin Invest, 1998, 102: 938
[32] 李小春,甘小丹.干燥综合征患者唇腺组织中三种细胞因子的检测,中国风湿病学杂志1999;3(3):148
[33] 丁大成,董怡,李申德等.干燥综合征病人小涎腺中细胞因子的表达,中华医学杂志;1996,76(6):427
[34] Horiuchi M, Yamano L, Inoue H, et al. Possible involvement of 11-12 express by Epstein-Barr vires in SjOgren's syndrome[J]. J Clin Pathol, 1999, 52: 833
[35] Barman SR, Berra A, Sterin-Borda L. Human primary SjOgren's syndrome autoantibodies asa mediators of nitric oxide release couple to lacrimal gland muscarinic acetylcholine receptors[J]. Curt Eye Res, 1998, 17: 1135
[36] Waterman SA, Gordon TP, Rischmueller M, et al. Inhibitory effects of muscarinic receptor autoantibodies on parasympathetic neurotransmission in SjOgren's syndrome[J]. Arthritis Rheum, 2000, 43: 1647
[37] 潘文奎.口眼干燥综合征的辨证论治.辽宁中医杂志,1988;5:21
[38] 马永桢.运用滋阴法治疗干燥综合征[J].甘肃中医,1995;8(1):13～14.1
[39] 王鹏宇,王静.辨证治疗干燥综合征73例[J].浙江中医杂志,1997;32(4):164～165.
[40] 陈淑琼,孙孝洪.中医治疗干燥综合征1例体会[J].四川中医,1998;16(4):35
[41] 徐宜厚.大补地黄丸治疗11例干燥综合征报告[J].中医杂志,1990;(8):42～
[42] 傅宗翰.干燥综合征初探.中医杂志,1983,(8):4
[43] 张立亭.张鸣鹤辨证治疗干燥综合征经验[J].山东中医药大学学报,2000;24(2):120～121.[7]
[44] 孙素平,潘文萍,周翠英.试论干燥综合征以燥毒为本[J].山东中医杂志,2001;20(10):581～582.
[45] 秦长林.从干燥综合征看“燥必入血”的病变特点[J].山东中医杂志,2000;19(12):710
[46] 董振华.活血化瘀法在干燥综合征中的运用.北京中医,2001;(3):9
[47] 王燕青,刘学法,李达祥.从瘀论治干燥综合征探微.中医函授通讯,1998;17(4):16～17
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