三黄凝胶对大鼠耳廓复合痤疮模型p38MAPK信号通路及其关键分子的影响

Effects of Sanhuang Gel on p38MAPK Signaling Pathway and Its Key Molecules in Rat Auricle Acne Complex Model

关键词:  三黄凝胶  痤疮  p38MAPK信号通路  MCP-1  HMGB1  α-SMA

Key words:  Sanhuang gel  acne  p38MAPK signaling pathway  MCP-1  HMGB1  α-SMA

摘要:

目的 以p38MAPK信号通路为切入点，探讨三黄凝胶对大鼠耳廓复合痤疮模型的抗炎作用机制。方法 将62只Wistar雄性大鼠分为空白组与造模组。造模组复制痤疮实验动物模型(耳廓涂抹100%油酸+皮下注射痤疮丙酸杆菌菌液)，21 d后随机抽取2只做HE染色，剩余大鼠分为模型组、阳性对照组(维A酸乳膏1.48 g·d^-1)和三黄凝胶高、中、低(1.1、0.568、0.3 g·d^-1)剂量组，连续用药28 d，并在治疗第7，14，28天对各组大鼠耳廓外观进行观察评价；取右侧耳廓组织，HE染色观察组织病理学变化，RT-PCR检测p38MAPK、MKK3、MKK6 mRNA的表达，IHC和Western blotting检测耳廓组织中MCP-1、HMGB1、α-SMA蛋白的表达。结果 与空白组比较，模型组大鼠耳廓组织表皮角化严重，炎细胞浸润明显，p38MAPK、MKK3、MKK6 mRNA水平，MCP-1、HMGB1、α-SMA蛋白表达水平明显升高(P<0.05)。经药物干预后，各治疗组大鼠表观评分均有不同程度改善，三黄凝胶高、中剂量组和阳性对照组p38MAPK、MKK3、MKK6 mRNA及MCP-1、HMGB1、α-SMA蛋白水平均明显降低(P<0.05)；三黄凝胶低剂量组MKK3 mRNA和MCP-1、HMGB1蛋白表达明显下降(P<0.05)，但p38MAPK、MKK6 mRNA和α-SMA蛋白表达改变不明显。结论 三黄凝胶缓解痤疮丙酸杆菌菌液诱导的炎症反应状态，减少组织炎症损伤，可能与其干预p38MAPK蛋白反馈调节信号通路，抑制MCP-1、HMGB1、α-SMA表达有关。

Abstract:

OBJECTIVE To observe the anti-inflammatory mechanism of Sanhuang gel on rat model of auricle acne and its possible mechanism with p38MAPK signaling pathway as the entry point. METHODS Sixty-two male Wistar rats were divided into blank group and modelling group. The modeling group replicated the experimental animal model of acne(application of 100% oleic acid+subcutaneous injection of Propionibacterium acnes), and 2 rats were randomly selected for HE staining after 21 days. The remaining rats were randomly divided into model group, positive control group(Vitamin A cream 1.48 g·d^-1 ), Sanhuang gel high, medium and low-dose group(1.1, 0.568, 0.3 g·d^-1 ). They were administered for 8 weeks, and observed on the 7, 14 and 28 day of treatment. The right auricle tissues were collected, and the histopathological changes were observed by HE stain. The mRNA expressions of p38MAPK, MKK3 and MKK6 were detected by RT-PCR. The protein expressions of MCP-1, HMGB1 and α-SMA in auricle tissues were detected by IHC and Western blotting. RESULTS Compared with blank group, the epidermis of auricular tissue in model group was severely keratinized, inflammatory cell infiltration was obvious, mRNA levels of p38MAPK, MKK3, MKK6 and protein expression levels of MCP-1, HMGB1 and α-SMA were increased(P<0.05). After drug therapy intervention, auricle apparent score at different time points in each treatment group was improved to varying degrees. The mRNA levels of p38MAPK, MKK3, MKK6 and protein levels of MCP-1, HMGB1 and α-SMA in Sanhuang gel high-dose and medium-dose groups and positive control group were decreased(P<0.05), while in Sanhuang gel low-dose group, the expressions of MKK3 mRNA and MCP-1, HMGB1 protein were decreased(P<0.05), the expression of p38MAPK, MKK6 mRNA and α-SMA protein decreased not significantly. CONCLUSION The mechanism of the effect of Sanhuang gel on the inflammatory response of rat compound acne animal model may be related to its intervention of p38MAPK protein feedback regulation signaling pathway and inhibition of MCP-1, HMGB1 and α-SMA expression.