基于p38 MAPK通路探讨盘龙七片对膝骨关节炎模型大鼠的治疗作用与可能机制  

Therapeutic Effect and Mechanism of Panlongqi Tablet in Treating KOA Rats Based on p38 MAPK Pathway

作　　者：时超[1] 谭亮[1] 孙春生 陈继平[1] 孟建国 SHI Chao;TAN Liang;SUN Chunsheng;CHEN Jiping;MENG Jianguo(Xuzhou Hospital of Traditional Chinese Medicine Affiliated to Nanjing University of Chinese Medicine,Xuzhou 221003,China;Shaanxi Panlong Pharmaceutical Group Co.,Ltd.,Xi′an 710025,China)

机构地区：[1]南京中医药大学附属徐州市中医院,江苏徐州221003 [2]陕西盘龙药业集团股份有限公司,陕西西安710025

出　　处：《中医药信息》2021年第12期19-25,共7页Information on Traditional Chinese Medicine

基　　金：陕西省重点研发计划项目(2020ZDLSF03-10)。

摘　　要：目的:探讨盘龙七片通过调控p38丝裂原活化蛋白激酶(p38 MAPK)通路对膝骨关节炎(KOA)大鼠的治疗作用。方法:60只SD大鼠随机分为正常对照组、模型组、盘龙七片组、p38 MAPK抑制剂组(SB203580组)、p38 MAPK激动剂组(茴香霉素组)和盘龙七片+茴香霉素组,每组各10只。除正常对照组外,其余各组大鼠采用膝关节穿刺注射胶原酶法制备KOA模型。造模成功后各治疗组分别给予相应药物,连续给药14 d。给药结束后观察各组大鼠行为学并评分;检测膝关节肿胀程度;采用HE染色法观察软骨组织病理变化;采用ELISA法检测软骨组织白细胞介素-1β(IL-1β)、肿瘤坏死因子(TNF-α)水平;采用免疫组化法检测磷酸化p38 MAPK(p-p38 MAPK)、磷酸化核转录因子κB(p-NF-κB)在软骨组织中的表达;采用WesternBlot法测Ⅱ型胶原(COL2)、解聚蛋白样金属蛋白酶-4(ADAMTS-4)、基质金属蛋白酶-13(MMP-13)、水通道蛋白3(AQP3)和凋亡蛋白(Caspase-3)的表达情况。结果:造模后,模型组大鼠右后肢痉挛收缩、行走困难等KOA行为学评分增加,软骨组织坏死、纤维组织增生、炎性浸润等病理损伤严重,软骨组织IL-1β及TNF-α水平升高,p38 MAPK/NF-κB/AQP3途径活化(P<0.05);盘龙七片组及SB203580组大鼠的软骨组织炎性病变及右后肢痉挛收缩、行走困难症状均明显缓解,p38 MAPK/NF-κB/AQP3通路及相关蛋白表达均受抑制(P<0.05);茴香霉素组大鼠KOA的相关症状加重(P<0.05);盘龙七片+茴香霉素组中茴香霉素则会逆转盘龙七片的上述作用(P<0.05)。结论:盘龙七片可通过抑制p38 MAPK/NF-κB/AQP3通路活化,缓解KOA大鼠软骨组织炎性损伤症状,可为盘龙七片治疗KOA提供理论依据。Objective:To explore the therapeutic effect of Panlongqi Tablet on rats with knee osteoarthritis(KOA)by regulating p38 mitogen-activated protein kinase(p38 MAPK)pathway.Methods:60 SD rats were randomly divided into the normal group,the model group,the Panlongqi Tablet group,the p38 MAPK inhibitor group(SB203580),the p38 MAPK agonist group(anisomycin),and the combination group(Panlongqi Tablet plus anisomycin),with 10 rats in each group.KOA model was prepared by knee injection of collagenase.After 14 days of continuous interventions,the scores of ethology and knee joint swelling were observed and assessed;HE method was used to detect pathological changes of cartilage tissue;ELISA method was applied to detect the levels of IL-1βand TNF-αin cartilage tissues;immunohistochemical method was used to measure the expressions of p-p38 MAPK and p-NF-κB in cartilage tissues;Western Blot method was used to detect the expressions of COL2,ADAMTS4,MMP-13,AQP3 and Caspase-3.Results:After modeling,the KOA behavioral scores were increased,such as spasticity and contraction of the right hind limbs and difficulty in walking,and severe pathological damages,such as cartilage tissue necrosis,fibrous tissue proliferation and inflammatory infiltration,could been observed;the levels of IL-1βand TNF-αin cartilage tissues were increased,p38 MAPK/NF-κB/AQP3 pathway was activated in the model group(P<0.05).The inflammatory lesions of cartilage tissues,the spasticity and contraction of right hind limb and the symptoms of walking difficulties were significant relieved,and the p38 MAPK/NF-κB/AQP3 pathway and related protein expression were inhibited in the Panlongqi Tablet group and the p38 MAPK inhibitor group(P<0.05).The symptoms of KOA were aggravated in the p38 MAPK agonist group(P<0.05).Anisomycin could reverse the above-mentioned effects of Panlongqi Tablet in the combination group(P<0.05).Conclusion:Panlongqi Tablet can inhibit the activation of p38 MAPK/NF-κB/AQP3 pathway and relieve the symptoms of inflammatory damages to the ca

关 键 词：盘龙七片 膝骨关节炎 软骨组织 p38丝裂原活化蛋白激酶通路 炎症 

分 类 号：R285.[医药卫生—中药学]