口炎清颗粒对阴虚型口腔溃疡模型大鼠的作用机制

Mechanism of Kouyanqing Granule on Oral Ulcer in Model Rats with Yin Deficiency

目的观察口炎清颗粒对阴虚型口腔溃疡模型大鼠溃疡愈合的作用,初步探讨口炎清颗粒养阴功效的作用机制。方法大鼠随机分为正常组、模型组、知柏地黄丸组(1.08 g·kg^(-1))及口炎清高、中、低剂量组(8,4,2 g·kg^(-1))。通过联合使用灌胃甲状腺素和Na OH烧灼法建立大鼠阴虚型口腔溃疡复合模型;观察大鼠的体质量、饮水量、体温等一般状况;采用ELISA法检测血浆环磷酸腺苷(cAMP)、环磷酸鸟苷(c GMP)含量,计算c AMP/c GMP比值,检测红细胞膜Na+-K+-ATP酶活性等指标;观察大鼠口腔溃疡的愈合情况;采用ELISA法检测模型大鼠口腔黏膜组织中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)、表皮生长因子(EGF)、转化生长因子-β1(TGF-β1)的含量;采用Western Blot法检测p38丝裂原活化蛋白激酶(p38MAPK)的表达。结果与正常组比较,模型组大鼠体质量减轻、饮水量增多、体温升高(P<0.05,P<0.01),大鼠溃疡修复评分显著降低(P<0.01),血浆c AMP水平、c AMP/c GMP比值及红细胞膜Na+-K+-ATP酶活性均明显升高(P<0.01),c GMP水平降低(P<0.05),口腔黏膜组织中TNF-α、IL-6含量明显升高(P<0.05),IL-10、EGF、TGF-β1含量明显降低(P<0.05,P<0.01),p38MAPK蛋白表达显著上升(P<0.05)。与模型组比较,口炎清中、高剂量组大鼠体质量明显增加(P<0.05,P<0.01);各给药组大鼠的饮水量均有所减少、体温下降(P<0.05,P<0.01),溃疡修复评分明显升高(P<0.05,P<0.01);口炎清高、中剂量组大鼠血浆c AMP水平、c AMP/c GMP比值及红细胞膜Na+-K+-ATP酶活性均明显降低(P<0.05,P<0.01),口炎清中剂量组大鼠血浆c GMP水平明显升高(P<0.05);口炎清高剂量组大鼠口腔黏膜TNF-α、IL-6含量均明显降低(P<0.05,P<0.01),口炎清高、中剂量组大鼠口腔黏膜IL-10、EGF、TGF-β1含量亦明显升高(P<0.01,P<0.05);口炎清各剂量组大鼠口腔黏膜组织匀浆中p38MAPK蛋白表达明显降低(P<0.05),且呈剂量相关性。结论口炎清颗粒能通过其"养阴"作用促进阴虚型口腔溃疡愈合;介导p38MAPK蛋白表达,抑制炎症反应,调节生长因子水平可能是口炎清颗粒发挥养阴功效的作用机制。

Objective To observe the effect of Kouyanqing granule on oral ulcer in the model rats with yin deficiency, and to investigate mechanism of the nourishing-yin effect of Kouyanqing granule. Methods The ratswere randomly divided into normal control group,model group,Zhi Bai Dihuang pills group（1.08 g·kg^（-1））,andthe high,medium and low dose of Kouyanqing granule groups（8,4,2 g·kg^（-1））. Thyroxine and Na OH were usedto induce the oral ulcer model rats with yin deficiency. The symptoms of yin deficiency including weight, waterintake,body temperature,and c AMP,c GMP,c AMP/c GMP in blood plasma,as well as Na＋-K＋-ATP enzymeactivity in erythrocyte membrane were measured. The effect of Kouyanqing on the degree of ulceration in model rats was observed. TNF-α,IL-6,IL-10,EGF,TGF-β1 contents in oral mucosa were measured by ELISA,andexpression of p38 MAPK in oral mucosa was also detected by Western Blot. Results Compared with the normalcontrol group,rats in model group showed significant decrease in weight,and increase in water intake and bodytemperature（P〈0.05,P〈0.01）. The score of ulcer repairement were reduced obviously（P〈0.01）. The modelgroup also showed significant increase in c AMP,c AMP/c GMP in blood plasma（P〈0.01）,Na＋-K＋-ATP enzymeactivity in erythrocyte membrane（P〈0.01）,TNF-α,IL-6 levels in oral mucosa（P〈0.05）,and the expressionof p38 MAPK（P〈0.05）. The levels of c GMP in blood plasma（P〈0.05） and IL-10,EGF,TGF-β1 in oralmucosa（P〈0.05,P〈0.01） were decreased obviously in model group. Compared with the model group,the highand medium dose of Kouyanqing granule showed significant increase in weight（P〈0.05,P〈0.01）. Kouyanqing granule could reduce water intake and body temperature（P〈0.05, P〈0.01）, and increase the score of ulcerrepaire（P〈0.05, P〈0.01） obviously. The high and medium dose of Kouyanqing granule could reduce c AMP,c AMP/c GMP levels,and Na＋-K＋-ATP enzyme activity（P〈0.05,P〈0.01）. The level of c GMP was increasedobviously（P〈0.05） in medium dose group of Kouyanqing granule. At the same time,the high dose of Kouyanqing granule could reduce TNF-α,IL-6 levels in oral mucosa remarkably（P〈0.05,P〈0.01）. The high and mediumdose of Kouyanqing granule could increase IL-10, EGF, TGF-β1 levels obviously（P〈0.05, P〈0.01）. Theexpression of p38 MAPK could be reduced by Kouyanqing granule（P〈0.05） in a dose-dependent manner.Conclusion Kouyanqing granule could improve the symptoms of yin deficiency. Kouyanqing granule has remarkableeffect on oral ulcer with yin deficiency. The role of Kouyanqing granule is related to the nourishing-yin effect via mechanisms which may be associated with mediating the expression of p38 MAPK to inhibit inflammatory reaction andelevating levels of EGFs.