罗哌卡因对骨关节炎疼痛大鼠炎症反应及PKA/CREB通路的影响

The Effects of Ropivacaine on Inflammatory Response and PKA/CREB Pathway in Rats with Osteoarthritis Pain

目的:探讨罗哌卡因对骨关节炎(OA)疼痛大鼠炎症反应及蛋白激酶A(PKA)/cAMP反应元件结合蛋白(CREB)通路的影响。方法:大鼠随机分为假手术组、OA组(建模)及罗哌卡因低、中、高剂量组(建模+腹腔注射0.5、1、2mg/kg罗哌卡因),每组16只,切断前交叉韧带建模2周后进行给药,给药两周。测定模型复制前、模型复制2周、给药2周时大鼠的压痛阈值,酶联免疫(ELISA)法检测血清IL-1β、IL-6、TNF-α水平,实时荧光定量PCR法测定软骨组织IL-1β、IL-6、TNF-α、PKA、CREB mRNA表达水平,Western Blot法测定软骨组织p-PKA/PKA、p-CREB/CREB水平,苏木精-伊红染色观察软骨组织病变。结果:给药2周,与假手术组相比,OA组大鼠压痛阈值显著降低(P<0.05),血清IL-1β、IL-6、TNF-α水平及软骨组织中IL-1β、IL-6、TNF-α、PKA、CREB mRNA表达水平、p-PKA/PKA、p-CREB/CREB蛋白表达水平显著升高(P<0.05),软骨组织出现明显的病变现象;与OA组相比,罗哌卡因低、中、高剂量组大鼠压痛阈值显著升高(P<0.05),血清IL-1β、IL-6、TNF-α水平及软骨组织中IL-1β、IL-6、TNF-α、PKA、CREB mRNA表达水平、p-PKA/PKA、p-CREB/CREB蛋白表达水平显著降低(P<0.05),且呈剂量依赖性,软骨组织病变减轻。结论:罗哌卡因可抑制OA疼痛大鼠PKA/CREB通路激活,降低体内炎症反应,减轻膝关节疼痛程度。

Objective:To investigate the effects of ropivacaine on the inflammatory response and protein kinase A(PKA)/cAMP response element binding protein(CREB)pathway in rats with osteoarthritis(OA)pain.Methods:Rats were randomly divided into sham operation group,OA group(modeling)and low,medium,and high dose ropivacaine groups(modeling+intraperitoneal injection of 0.5,1,2 mg/kg ropivacaine),16 in each group;the drug was administered after 2 weeks of modeling by cut off ligamenta cruciata,for two weeks.The tenderness threshold of rats was measured before model replication,2 weeks after model replication,and 2 weeks after administration.Enzyme-linked immunoassay(ELISA)method was used to detect serum IL-1β,IL-6,and TNF-αlevels,real-time fluorescent quantitative PCR method to determine the expression levels of IL-1β,IL-6,TNF-α,PKA,CREB mRNA in cartilage tissue,Western Blot to determine the levels of p-PKA/PKA and p-CREB/CREB in cartilage tissue,hematoxylin-eosin staining to observe cartilage tissue lesions.Results:After 2 weeks of administration,compared with the sham operation group,the tenderness threshold of rats in the OA group was significantly reduced(P<0.05),the serum IL-1β,IL-6,TNF-αlevels,and cartilage tissue IL-1β,IL-6,TNF-α,PKA,CREB mRNA expression levels,p-PKA/PKA,p-CREB/CREB protein expression levels were significantly increased(P<0.05),the cartilage tissue had obvious pathological phenomena;compared with the OA group,the tenderness threshold of rats in the low,medium and high dose ropivacaine groups was significantly increased(P<0.05),the serum IL-1β,IL-6,TNF-αlevels,and cartilage tissue IL-1β,IL-6,TNF-α,PKA,CREB mRNA expression levels,p-PKA/PKA,p-CREB/CREB protein expression levels were significantly reduced(P<0.05),and in a dose-dependent manner,the cartilage tissue lesions were alleviated.Conclusion:Ropivacaine can inhibit the activation of PKA/CREB pathway in rats with OA pain,reduce inflammation in the body,and reduce the degree of knee joint pain.