隐丹参酮调节HIF-1α/BNIP3信号通路对兔膝骨关节炎模型软骨细胞自噬和凋亡的影响  

Impacts of cryptotanshinone on autophagy and apoptosis of chondrocytes in rabbit model of knee osteoarthritis by regulating HIF-1α/BNIP3 signaling pathway

作　　者：王柯[1] 叶寒露[2] WANG Ke;YE Hanlu(Department of Orthopedics and Traumatology,Wuhan Traditional Chinese Medicine Hospital,Wuhan 430014,China;Department of Endocrinology,Wuhan Traditional Chinese Medicine Hospital,Wuhan 430014,China)

机构地区：[1]武汉市中医医院骨伤科,430014 [2]武汉市中医医院内分泌科,430014

出　　处：《天津医药》2024年第4期372-378,共7页Tianjin Medical Journal

基　　金：武汉市中医药科研项目(wz22c62)。

摘　　要：目的 探究隐丹参酮调节缺氧诱导因子-1α(HIF-1α)/腺病毒E1B19kDa相互作用蛋白3(BNIP3)信号通路对兔膝骨关节炎(KOA)模型软骨细胞自噬和凋亡的影响。方法 取新西兰兔并以改良Videman法构建兔KOA模型,随机分为模型组、空载组、隐丹参酮组、HIF-1α敲低组、隐丹参酮+HIF-1α敲低组,每组9只;另取9只新西兰兔为对照组。分组干预后以Lequesne MG的膝关节级别评估法对兔膝关节临床症状(局部疼痛、步态、关节活动、关节肿胀)进行评分;HE染色检测兔膝关节软骨组织的退变情况并进行改良Mankin's评分;TUNEL染色检测兔膝关节软骨组织细胞凋亡情况;酶联免疫吸附试验(ELISA)检测兔血清炎性因子白细胞介素(IL)-6、IL-18、IL-10水平;蛋白免疫印迹实验检测兔膝关节软骨组织自噬(LC3、Beclin-1)、凋亡(Bax、Cleaved Caspase-3)和HIF-1α/BNIP3信号通路相关蛋白表达。结果 与对照组比较,模型组兔膝关节软骨组织出现明显退变症状,局部疼痛、步态、关节活动及关节肿胀评分、改良Mankin's评分、凋亡率、血清IL-18及IL-6水平、软骨组织LC3Ⅱ/LC3Ⅰ、Beclin-1、Bax、Cleaved Caspase-3、BNIP3蛋白表达水平升高,血清IL-10水平、软骨组织HIF-1α蛋白表达水平降低(P<0.05)。与模型组比较,隐丹参酮组兔膝关节软骨组织退变症状减轻,局部疼痛、步态、关节活动及关节肿胀评分、改良Mankin's评分、凋亡率、血清IL-18及IL-6水平、软骨组织LC3Ⅱ/LC3Ⅰ、Beclin-1、Bax、Cleaved Caspase-3、BNIP3蛋白表达水平降低,血清IL-10水平、软骨组织HIF-1α蛋白表达水平升高(P<0.05);HIF-1α敲低组兔膝关节软骨组织退变症状加重,局部疼痛、步态、关节活动及关节肿胀评分、改良Mankin's评分、凋亡率、血清IL-18及IL-6水平、软骨组织LC3Ⅱ/LC3Ⅰ、Beclin-1、Bax、Cleaved Caspase-3、BNIP3蛋白表达水平升高,血清IL-10水平、软骨组织HIF-1α蛋白表达水平降低(P<0.05);空Objective To investigate the impact of cryptotanshinone on autophagy and apoptosis of chondrocytes in a rabbit knee osteoarthritis(KOA)model by regulating the hypoxia inducible factor-1α(HIF-1α)/BCL2 and adenovirus E1B 19-kDa-interacting protein 3(BNIP3)signaling pathway.Methods New Zealand rabbits were selected to construct the rabbit KOA model using the improved Videman method.Rabbits were randomly divided into the model group,the empty group,the cryptotanshinone group,the HIF-1αknockdown group and the cryptotanshinone+HIF-1αknockdown group,with 9 rabbits in each group.Another 9 New Zealand rabbits were taken as the control group.After grouping and intervention,Lequesne MG knee joint level assessment method was used to score clinical symptoms of knee joints(local pain,gait,joint activity and joint swelling)of rabbits.HE staining was used to detect the cartilage degeneration of rabbit knee joint cartilage tissue,and modified Mankin's score was performed.TUNEL staining was used to detect the apoptosis of articular cartilage histiocytes of rabbits.Enzyme-linked immunosorbent assay(ELISA)method was used to detect serum levels of inflammatory cytokines interleukin-6(IL-6),IL-18 and IL-10 in rabbits.Western blot experiment was used to detect the expression of autophagy(LC3,Beclin-1),apoptosis(Bax,Cleaved Caspase-3)and HIF-1α/BNIP3 signaling pathway related proteins in knee cartilage tissue of rabbits.Results Compared with the control group,the knee joint cartilage tissue of rabbits showed obvious degenerative symptoms,the scores of local pain,gait,joint activity and joint swelling,the modified Mankin's score,apoptosis rate,levels of serum IL-18 and IL-6,and the expression of LC3Ⅱ/LC3Ⅰand Beclin-1,Bax,Cleaved Caspase-3,BNIP3 proteins in cartilage tissue were increased in the model group,and the serum level of IL-10 and the expression of HIF-1αprotein in cartilage tissue were decreased(P<0.05).Compared with the model group,the symptoms of cartilage tissue degeneration in knee joint of rabbits were alleviated,

关 键 词：隐丹参酮 骨关节炎 膝 软骨细胞 自噬 凋亡 HIF-1α/BNIP3 

分 类 号：R285.5[医药卫生—中药学]