【作者】 洪志明； 陈德宁； 曾杨玲； 周文彬； 丁晓虹； 王全；

【Author】 HONG Zhiming;CHEN Dening;ZENG Yangling;ZHOU Wenbin;DING Xiaohong;WANG Quan;Shenzhen TCM Hospital;Guangzhou General Hospital of Guangzhou Military Area Command of Chinese PLA;

【机构】 深圳市中医院； 广州军区广州总医院；

【摘要】 目的:观察前痛定方对慢性前列腺炎(CP)疼痛动物模型脊髓小胶质细胞p38丝裂原活化蛋白激酶(p38MAPK)信号转导通路的影响,探寻前痛定方抗炎、止痛作用的机制和靶点所在。方法:80只雄性SD大鼠随机分成4组:A组(正常组,n=20)、B组(假造模组,n=20)、C组(前痛定组,n=20)、D组(蒸馏水组,n=20)。A组正常饲养;B组采用前列腺腹侧叶注射生理盐水0.1ml假造模,C组、D组前列腺腹侧叶注射弗氏完全佐剂(FCA)0.1ml造模CP。造模后第5天开始,A组、B组、D组蒸馏水1ml/100g灌胃;C组前痛定方溶液1ml/100g灌胃,每天一次。分别于造模后第5天、第11天、第18天、34天,每组抓出5只大鼠,采用热辐射刺激仪测定大鼠反射潜伏期(TWL);脱臼处死后取出前列腺组织送病理检查;实时荧光定量反转录-聚合酶链反应(RT-PCR)法检测CP模型大鼠L5-S2段脊髓IBA-1 mRNA、p38MAPK m RNA的表达,Western Blot法检测L5-S2段脊髓p38MAPK、磷酸化p38丝裂原活化蛋白激酶(pp38MAPK)蛋白的表达。结果:造模后第5天,C组TWL值明显缩短,前列腺腺体结构破坏,腺腔扩张,多数腺腔内可见组织细胞、炎细胞和变性坏死组织等,间质水肿,小血管扩张充血,间质及腺体内灶状淋巴细胞浸润或淋巴细胞浸润呈片状分布;L5-S2段脊髓内IBA-1 mRNA、p38MAPK mRNA、pp38MAPK蛋白的表达水平明显上调。造模后第11天、第18天、第34天,C组TWL值逐渐延长,前列腺组织炎症逐渐消退,L5-S2段脊髓内IBA-1mRNA、p38MAPK mRNA、pp38MAPK蛋白的表达水平逐渐下调。结论:前列腺腹侧叶注射FCA复制CP模型成功,前痛定方对CP模型大鼠有明显的抗炎、止痛效果,下调L5-S2段脊髓内p38MAPK信号转导通路的信号转导、抑制小角质细胞的活化,可能是前痛定方发挥抗炎、止痛作用的机制和靶点所在。更多还原

【Abstract】 Objective:To observe the influence of Qiantongding Decoction on p38 MAPK signal transduction pathways of spinal microglia in CP pain animal models.Methods: 80 male SD rats were randomized into Group A(normal group, n=20), Group B(sham modeling group, n=20), Group C(Qiantongding Group,n=20) and Group D(Water Group, n=20). Group A were normally access to food. Group B were applied sham modeling, with normalsaline injection(0.1 ml) to the ventral lobes of the prostates.Group C and Group D were applied Freund’s complete adjuvant(FCA)injection(0.1 ml) to the ventral lobes of the prostates to establish CP models. From the 5 thday following surgeries, Group A, Group B and Group D were intragastrically administered 1 ml/100 g water, and rats in Group C were intragastrically administered 1 ml/100 g Qiantongding Solution, once a day. On the fifth, eleventh, eighteenth and thirty-fourth day following the surgeries, five rats in each group were radiated to the pelma with the heat radiation stimulus to determine Thermal Withdrawal Latency(TWL). After scarified, the prostates and L5-S2 medulla spinalis were removed. Prostates were observed under light microscopy. The IBA-1 mRNA and p38 MAPK mRNA protein expressions in the L5-S2 medulla spinalis of CP rats were detected with real-time fluorescence quota PCR;the p38 MAPK and pp38 MAPK protein expressions in the L5-S2 medulla spinalis were detected with Western Blotting.Results:On the 5 thday following surgeries, the TWL values in Group C were obviously decreased(P<0.05). The structures of the prostate glands were destroyed, with the glandular cavities expanded, and histocytes, inflammatory cells and degenerated and necrotic tissues filling in most of the glandular cavities,with the mesenchyma edema, small vessels expanded and congested,and lymphocyte infiltration in the mesenchyma and glands, or with patchy distribution of the lymphocyte infiltration. The expressions of IBA-1 mRNA, p38 MAPK mRNA and pp38 MAPK proteins in the L5-S2 medulla spinalis were increased significantly(P<0.05). On the 11 th, 18 thand 34 thday following surgeries, the TWL values were increased gradually. The inflammation of the prostates alleviated gradually. The expressions of IBA-1 mRNA,p38 MAPK m RNA and pp38 MAPK proteins in the L5-S2 medullaspinalis were decreased gradually. Conclusions: The CP modeling establishment with CFA injection to the ventral lobes of the prostates was successful. Qiantongding Decoction has the obvious effects of anti-inflammation and pain alleviation to the CP model rats. Qiantongding Decoction could inhibit the signal transduction of p38 MAPK signal transduction pathways,inhibit the activation of the microglia, in the L5-S2 medulla spinalis, hence performing the actions of anti-inflammation and pain alleviation.更多还原