摘要
目的探讨甘草甜素对膝关节骨性关节炎(KOA)大鼠软骨细胞凋亡及第10号染色体同源丢失性磷酸酶张力蛋白基因(PTEN)/蛋白激酶B(AKT)/环氧合酶2(COX-2)信号通路的影响。方法80只雄性SD大鼠分为假手术组、KOA组(造模)、阳性药物组(造模+灌胃6.75 mg/kg双氯芬酸钠)及低、高剂量甘草甜素组(造模+灌胃50、100 mg/kg甘草甜素),每组16只;假手术组大鼠手术不造模、后4组大鼠采用Hulth法造KOA模型,前2组大鼠术后2周灌胃生理盐水、后3组大鼠灌胃相应药物,共4周;于术前、术后2周、给药2、4周时测量各组大鼠右膝关节直径,ELISA法检测血清中白细胞介素1β(IL-1β)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)水平,HE染色法观察软骨组织形态,TUNEL染色法检测软骨组织细胞凋亡,Western blot法检测软骨组织PTEN、p-AKT/AKT、COX-2、兔抗人单克隆抗体(Bax)、活化型半胱天冬酶-3(cleaved caspase-3)蛋白表达。结果与假手术组相比,KOA组大鼠膝关节直径、血清IL-1β、IL-6、TNF-α水平及软骨组织细胞凋亡率、p-AKT/AKT、COX-2、Bax、cleaved caspase-3蛋白水平均升高(P<0.05),PTEN蛋白水平降低(P<0.05),软骨组织炎症病变重;与KOA组相比,低、高剂量甘草甜素组及阳性药物组大鼠膝关节直径、血清IL-1β、IL-6、TNF-α水平及软骨组织细胞凋亡率、p-AKT/AKT、COX-2、Bax、cleaved caspase-3蛋白水平均降低(P<0.05),PTEN蛋白水平升高(P<0.05),软骨组织炎症病变减轻,且高剂量甘草甜素组各项指标优于低剂量甘草甜素组、阳性药物组(P<0.05)。结论甘草甜素可能通过上调软骨组织中PTEN表达,抑制AKT/COX-2通路激活,减少软骨细胞凋亡,从而改善KOA病情。
Objective To investigate the effect of glycyrrhizin on chondrocyte apoptosis and phosphatase and tensin homolog(PTEN)/protein kinase B(AKT)/cyclooxygenase 2(COX-2)signaling pathway in rats with knee osteoarthritis(KOA).Methods Eighty male SD rats were divided into sham operation group(Sham),KOA group(modeling),positive drug group(modeling+gavage of 6.75 mg/kg diclofenac sodium),low-dose(modeling+50 mg/kg glycyrrhizin),and high-dose(modeling+100 mg/kg glycyrrhizin)groups.Each group had 16 rats.Except Sham group,Hulth method was performed on other 4 groups to establish KOA model.After operation,sham and KOA groups were gavaged with normal saline for 2 weeks,while positive drug group,low-and high-dose groups were gavaged with corresponding drugs for 4 weeks.The diameters of the right knee joint of the rats were measured before operation,2 weeks after operation,2 and 4 weeks after drug administration.The levels of serum interleukin 1β(IL-1β),interleukin 6(IL-6),and tumor necrosis factorα(TNF-α)were detected by ELISA.The morphology Cartilage tissues was observed by HE staining.TUNEL staining was applied to detect chondrocyte apoptosis.Western blot was used to detect the protein expression levels of PTEN,p-AKT/AKT,COX-2,Bax,activated caspase-3(cleaved caspase-3)in cartilage tissues.Results When compared to sham group,the knee joint diameter,the levels of serum IL-1β,IL-6,and TNF-α,chondrocyte apoptosis rate,the protein expression levels of p-AKT/AKT,COX-2,Bax and cleaved caspase-3 protein were increased(P<0.05),while PTEN level was decreased(P<0.05).Cartilage tissue inflammation became severe relative to sham group.When compared to KOA group,the knee joint diameter,the levels of serum IL-1β,IL-6,and TNF-α,apoptosis rate of cartilage tissue,the protein expression levels of p-AKT/AKT,COX-2,Bax,and cleaved caspase-3 were decreased in low-and high-dose glycyrrhizin groups and positive drug group(P<0.05),while PTEN level was increased(P<0.05).Cartilage tissue inflammation was alleviated relative to KOA group.The ind
作者
任天琦
刘泽霖
胡海
刘宏鹏
李小冬
REN Tianqi;LIU Zelin;HU Hai;LIU Hongpeng;LI Xiaodong(Department of Orthopedic Surgery,the Second Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150001,Heilongjiang,China)
出处
《贵州医科大学学报》
CAS
2022年第7期785-790,795,共7页
Journal of Guizhou Medical University
基金
黑龙江省中医药学会青年中医药科技创新计划项目(ZHY19-042)
黑龙江中医药大学科研基金项目(2019MS25)。
