独活寄生汤对膝骨关节炎模型大鼠PERK/Bip信号通路的影响  

Effect of Duhuo Jisheng Tang on PERK/Bip Signaling Pathway in Knee Osteoarthritis Model Rats

作　　者：赵林灿 李薇薇[2] 吴毅明[2] 段卫峰[3] ZHAO Lin-can;LI Wei-wei;WU Yi-ming;DUAN Wei-feng(Zhengzhou Shuqing Medical College, Zhengzhou 450000;The Third Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou 450004,China;Luoyang Orthopedic-traumatological Hospital of Henan Province,Luoyang 471000,China)

机构地区：[1]郑州澍青医学高等专科学校,郑州450000 [2]河南中医药大学第三附属医院,郑州450004 [3]河南省洛阳正骨医院,河南洛阳471000

出　　处：《中国实验方剂学杂志》2019年第9期18-24,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河南省中医药科学研究专项(2018ZY2018)

摘　　要：目的:基于蛋白激酶R样内质网激酶(PERK)/免疫球蛋白结合蛋白(Bip)信号通路探讨独活寄生汤缓解膝骨关节炎的可能机制。方法:采用寒冷刺激法建立膝骨性关节炎模型,将大鼠随机分为空白组,模型组,塞来昔布组(0.021 g·kg^(-1))和独活寄生汤低、中、高剂量组(8.37,16.72,33.48 g·kg^(-1)),空白组和模型组给予等体积的生理盐水,记录各组大鼠膝关节直径变化。采用苏木素-伊红(HE)染色观察大鼠膝关节软骨病理形态变化;酶联免疫吸附测定(ELISA)检测血清中肿瘤坏死因子-α(TNF-α),白细胞介素-1β(IL-1β),透明质酸(HA)的表达;采用实时荧光定量PCR(Real-time PCR)和蛋白免疫印迹法(Western bolt)检测软骨中PERK,Bip,半胱氨酸天冬氨酸蛋白酶-9(Caspase-9)mRNA和蛋白的表达。结果:与模型组比较,塞来昔布组和独活寄生汤高剂量组大鼠膝关节红肿明显改善,且关节直径均显著的减小(P<0.01),塞来昔布组和独活寄生汤高剂量组大鼠膝关节软骨局部裂隙缺损有明显好转。与空白组比较,模型组大鼠TNF-α,IL-1β和HA表达水平显著升高(P<0.01),大鼠软骨中PERK,Bip,Caspase-9的mRNA和蛋白表达水平明显升高(P<0.05,P<0.01);与模型组比较,塞来昔布组和独活寄生汤高剂量组大鼠血清中TNF-α,IL-1β和HA表达明显下降(P<0.05,P<0.01),塞来昔布组和独活寄生汤高剂量组大鼠软骨中PERK,Bip,Caspase-9的mRNA和蛋白表达水平明显降低(P<0.05,P<0.01)。结论:独活寄生汤能缓解膝骨关节炎模型大鼠的症状,其机制可能与调节大鼠软骨中PERK/Bip信号通路有关。Objective: To explore the possible mechanism of Duhuo Jisheng Tang in relieving knee osteoarthritis based on protein kinase R-like endoplasmic reticulum kinase( PERK)/immunoglobulin-binding protein( Bip) signaling pathway. Method: A model of knee osteoarthritis was established by cold stimulation.Rats were randomly divided into blank group,model group,celecoxib group( 0. 021 g·kg^-1),low,medium and high-dose Duhuo Jisheng Tang groups( 8. 37,16. 72,33. 48 g·kg^-1). Blank group and model group were given equal volume of physiological saline. The changes of knee joint diameter were recorded. The pathological changes of rat articular cartilage were observed by hematoxylin-eosin( HE) staining. The expressions of tumor necrosis factor-alpha( TNF-α),interleukin-1β( IL-1β) and hyaluronic acid( HA) in serum were detected by enzymelinked immunosorbent assay( ELISA). The mRNA and protein expression levels of PERK,Bip and cysteinyl as parates pecific protein-9( Caspase-9) in cartilage were detected by Real-time PCR and Western blot. Result:The knee joint redn ess and the joint diameter of celecoxib group and high-dose Duhuo Jisheng Tang group were improved,and the joint diameter was reduced significantly( P<0. 01). The local fractures of knee joint cartilage in the celecoxib group and the high-dose Duhuo Jisheng Tang group were significantly improved. Compared with blank group,the expression levels of TNF-α,IL-1β and HA were increased in model group( P< 0. 01). The gene and protein expression levels of PERK,Bip and Caspase-9 in cartilage of the model group were increased( P <0. 05,P <0. 01). Compared with model group,the expressions of TNF-α,IL-1β and HA in serum of celecoxib group and high-dose Duhuo Jisheng Tang group were decreased( P< 0. 05,P <0. 01). The mRNA and protein expression levels of PERK,Bip and Caspase-9 in cartilage of celecoxib group and high-dose Duhuo Jisheng Tang group were decreased( P< 0. 05,P <0. 01). Conclusion: Duhuo Jisheng Tang can alleviate the symptoms of knee osteoarthritis model rats,

关 键 词：独活寄生汤 骨性关节炎 蛋白激酶R样内质网激酶(PERK) 免疫球蛋白结合蛋白(Bip) 信号通路 

分 类 号：R289[医药卫生—方剂学] R684.3[医药卫生—中药学]