蜜炙远志降低胃粘膜损伤的机理研究

【作者】 刘贤武；

【导师】 王建；

【作者基本信息】 成都中医药大学 ， 中药学， 2008， 博士

【摘要】 目的:阐明蜜炙远志降低胃粘膜损伤的生物学机理,拟从分子水平揭示中药炮制降低胃肠毒性的科学内涵,对指导临床安全合理使用远志具有实用价值和学术意义。方法:应用药理学、胃肠电生理学、分子生物学等方法和手段,探索了生远志及其皂苷与蜜远志对胃粘膜损伤和保护机制。①采用甲基橙法考察了各受试药对小鼠胃排空的影响。②采用BL-420生物机能实验系统考察了对大鼠胃电生理的影响。③采用微循环实验观测了对小鼠胃粘膜血液循环的影响。④采用放免法测定了大鼠胃组织的P物质（SP）、生长抑素（SS）。⑤采用荧光分光光度法测定大鼠胃组织的前列腺素E₂（PGE₂）;采用定磷法测定了大鼠胃组织钙离子三磷酸腺苷(Ca⁺⁺-ATPase)酶活性。⑥采用RT-PCR法考察了大鼠胃粘膜环氧合酶2（COX-2）、钙调素（CaM）、转化生长因子α（α-TGF）、肠三叶因子（ITF）表达水平。结果:①生远志组与总皂苷组均能显著抑制小鼠胃排空（P＜0.01）,并能显著抑制胃电快波、慢波频率（P＜0.01,P＜0.05）;蜜远志则对小鼠胃排空、胃电快慢波均无显著影响。②生远志组能显著降低大鼠胃组织SP和SS的含量（P＜0.05）;而蜜远志组与总皂苷则显著增加胃组织SP含量,降低SS含量,皂苷尤为显著（P＜0.05）。③生远志组能显著降低小鼠胃粘膜血液流速、使毛细血管收缩、减少单位面积的血管数、减少单位面积的每秒血流量（P＜0.05）;蜜远志则能显著增加小鼠胃粘膜的血液流速、单位面积的每秒血流量（P＜0.05）。④生远志组及其皂苷组、蜜远志组有降低大鼠胃组织前列腺素E₂趋势,但无统计学意义。蜜远志组能显著增强大鼠胃粘膜COX-2的表达（P＜0.05）,而生远志及其总皂苷对COX-2表达无明显影响。⑤生远志能显著降低大鼠胃组织Ca⁺⁺-ATP酶活力（P＜0.05）,蜜远志及皂苷对其无显著影响。但蜜远志能显著增强钙调素基因的表达（P＜0.05）,生远志与皂苷组对其影响不明显。⑥蜜远志能显著增强大鼠胃粘膜ITF的表达（P＜0.05）,并能上调胃粘膜α-TGF基因表达,生远志对其无显著影响。结论:生远志10g/kg及其总皂苷5g/kg均能导致胃粘膜损伤,而蜜炙远志10g/kg则能降低粘膜损伤,其机理可能与:①通过增加胃电快波频率,促使胃组织中SP物质释放,从而增加胃排空,进而防止胃粘膜损伤。②通过增加胃粘膜血液流速、扩张毛细血管平滑肌、提高每秒血流量,从而改善胃粘膜微循环,提高胃粘膜的抗损伤和修复能力。③通过增强胃组织Ca²⁺-ATP酶活,消除细胞“钙超载”对粘膜的损害。④促进上皮细胞的分裂,加强对损伤部位的上皮整复。⑤增强胃粘膜肠三叶因子、α-TGF的表达,以增强对胃粘膜的保护和损伤修复等机制有关。综上,蜜炙远志通过促进胃粘膜微循环、增加胃组织局部激素释放、促使相关酶活性、增强粘膜保护因子的表达等多层次、多途径来实现胃粘膜的保护及损伤修复。更多还原

【Abstract】 Objective:Expound the biological mechanism of Radix Polygalae reducing the gastric mucosa damage by honey-fried,reveal the scientific connotation of traditional Chinese medicine reducing the stomach and intestines toxic by concoct in molecular level,It has great practical value and the academic significance of using Radix Polygalae reasonably and safely.Methods:Application method of Pharmacology、stomach and intestines electrophysiology、molecular biology and so on.Explored the crude Radix P.and its saponins and the honey-stir-baked Radix P.to the gastric mucosa damage and the protection mechanism,We did the following researches:①use helianthin to research gastric emptying;②use BL-420 biologic analyzer to observe and record the electrophysiology and gastric microcirculation;③use RIA to determine SP、SS;④use fluorescence pectrophotometriy to determine PGE₂,test enzyme activity of Ca⁺⁺-ATPase by the colorimetric determination of phosphorus;⑤use RT-PCR to study the expression of COX-2、CaM、α-TGF and ITF.Result:①Crude Radix P.and its saponins can obviously inhibit gastric motility（P＜0.01）,remarkably both inhibit stomach’s fast wave and slow wave（P＜0.01,P＜0.05）,but Honey-stir-baked Radix P.can not;②Crude Radix P.can obviously decrease SP and SS（P＜0.05）,Honey-stir-baked Radix P.can remarkably increase SP and decrease SS;③Crude Radix P.can remarkably cause gastric mucosal microcycle barrier（P＜0.05 ）;④Both Crude Radix P.and Honey-stir-baked Radix P. have not influence to PGE2;Honey-stir-baked Radix P.can enhance cox-2 expression （P＜0.05）;⑤Crude Radix P.can obviously decrease the activity of Ca⁺⁺-ATPase in gastric tissue and Honey-stir-baked Radix P.enhance CaM expression（P＜0.05）;⑥Honey-stir-baked Radix P.can enhanceα-TGF and ITF expression（P＜0.05）.Conclusion:Crude Radix P.（10g/kg） and its saponins（5g/kg） can damage the gastric mucosa,but Honey-stir-baked Radix P.can reduce this damage,The biological mechanism of Honey-stir-baked Radix P.reducing the gastric mucosal damage may related to the following factors:①Honey-stir-baked Radix P.can obviously increase SP of gastric tissue and improve the suppression of Crude Radix P. to stomach’s fast wave,Thus improves the gastric motility;②Increase gastric mucosal blood flow,enhances the anti-damage and repair ability;③Improve the suppression of Crude Radix P.to the activity of Ca⁺⁺-ATPase in gastric tissue, eliminates the cell "calcium overload" to the mucous membrane harm;④The promotion epithelial cell’s fission and strengthening restitution of damaging region;⑤Enhancea-TGF and ITF expression,increase the gastric mucosa protection and damage repair.In brief,Honey-stir-baked Radix P.realize multi-level,the multi-ways to the gastric mucosa protection and the damage repair mainly through the promoting gastric mucosa microcycle,enhancing stomach organization local hormone release、urged the related enzyme activity、urged the gastric mucosal protection factor expression and so on.更多还原