黄芩总黄酮对大鼠膝骨性关节炎软骨组织Camk2d、Ppp3r2表达的影响

Effect of total flavonoids of Scutellaria baicalensis Georgi on Camk2d and Ppp3r2 expression in cartilage of knee osteoarthritis rats

目的:探讨黄芩总黄酮对大鼠膝骨性关节炎(knee osteoarthritis,KOA)软骨组织钙/钙调蛋白依赖性蛋白激酶Ⅱδ(Camk2d)、蛋白磷酸酶3调节亚基2(Ppp3r2)表达的影响。方法:采用膝关节腔内注射木瓜蛋白酶的方法制备大鼠KOA模型,阴性对照组和模型组大鼠灌胃给予生理盐水,黄芩总黄酮低、中和高剂量组大鼠灌胃给予黄芩总黄酮,剂量分别为12.5、25、50 mg/kg,阳性对照组大鼠灌胃给予塞来昔布(10 mg/kg),连续14 d后对大鼠膝关节进行病理学观察和Makin′s评分,同时ELISA法检测膝关节软骨组织中肿瘤坏死因子-α(TNF-α)、IL-1β、IL-6含量,蛋白质印迹法检测膝关节软骨组织中Camk2d和Ppp3r2表达水平。结果:模型组大鼠膝关节软骨纤维结缔组织增生,结构不清,伴炎症细胞浸润;黄芩总黄酮低剂量组大鼠膝关节软骨纤维结缔组织增生,细胞排列无规则,炎症细胞浸润,但较模型组明显改善,随着黄芩总黄酮剂量的增加,病变改善越明显。与阴性对照组比较,模型组大鼠膝关节软骨组织Makin′s评分,TNF-α、IL-1β、IL-6和Camk2d水平明显升高,Ppp3r2水平明显降低(P均<0.05);与模型组相比,黄芩总黄酮各剂量组及阳性对照组大鼠膝关节软骨组织Makin′s评分,TNF-α、IL-1β、IL-6和Camk2d水平明显降低,Ppp3r2水平明显升高,且黄芩总黄酮各剂量组效应呈剂量依赖性(P均<0.05);黄芩总黄酮高剂量组和阳性对照组间各指标差异无统计学意义(P均>0.05)。结论:黄芩总黄酮可能通过增加软骨组织中Ppp3r2表达,降低Camk2d表达,对大鼠KOA发挥治疗作用。

Objective:To investigate the effect of total flavonoids of Scutellaria baicalensis Georgi(TFSB)on calcium/calmodulin dependent protein kinaseⅡdelta(Camk2d)and protein phosphatase 3 regulates subunit 2(Ppp3r2)expression in cartilage of knee osteoarthritis rats.Methods:The knee osteoarthritis model was prepared by injecting papain into the knee joint cavity.The rats in the negative control group and the model group were given normal saline by intragastric administration.Rats in TFSB low-dose group,middle-dose group and high-dose group were given the TFSB by gavage at the doses of 12.5,25,and 50 mg/kg.The positive control group rats were gavaged with celecoxib(10 mg/kg),and treated for 14 consecutive days.The pathological knees joint of rats was observed and Makin′s score were performed,and the TNF-α,IL-1β,IL-6,Camk2d and Ppp3r2 of knee cartilage tissue were detected by ELISA.The levels of Camk2d and Ppp3r2 protein in knee cartilage tissue were detected by Western Blotting.Results:In the model group,the knee cartilage fiber connective tissue was proliferated,and the structure was unclear,and the inflammatory cells infiltrated.The low-dose group of TFSB had hyperplasia of articular cartilage fiber connective tissue,irregular cell arrangement and inflammatory cell infiltration,but it was significantly improved compared with the model group.With the increasing of the dose of TFSB,the improvement of the lesion was more obvious.Compared with the negative control group,the Makin′s score,TNF-α,IL-1β,IL-6 and Camk2d levels in the knee cartilage tissues of the model group increased,while the Ppp3r2 level decreased(all P<0.05).After the TFSB or celecoxib treatment,compared with the model group,the Makin′s score,TNF-α,IL-1β,IL-6 and Camk2d levels in each dose group of TFSB and the positive control group decreased,and the level of Ppp3r2 increased,and the effect of each dose group of TFSB was dose-dependent(all P<0.05).The indicators between the high-dose group of TFSB and the positive control group were no significant difference(all P>0.05).Conclusion:TFSB may exert therapeutic effect in rats with knee osteoarthritis by increasing the expression of Ppp3r2 and reducing the expression of Camk2d in cartilage.