丹皮酚通过调节Wnt途径对强直性脊柱炎小鼠疗效及滑膜组织中RANKL和Smadl水平的影响

Effects of Paeonol on the efficacy of AS mice and the levels of RANKL and Smalll in synovial tissue by regulating Wnt pathway

目的分析丹皮酚对强直性脊柱炎(AS)小鼠治疗疗效及对Wnt通路与RANKL和Smadl水平的影响。方法取100只小鼠随机分为模型组、阳性药物组、空白组、丹皮酚组;除空白组外,其余三组均使用100μl蛋白聚糖+100μl弗氏完全佐剂建立强直性脊柱炎模型;造模成功后开始给药,阳性药物组给予柳氮磺吡啶9 mg/kg进行灌胃,丹皮酚组给与丹皮酚3 mg/kg进行灌胃,模型组及空白组给予等体积的蒸馏水。各组每天给药1次,连续进行20 d。在造模前及造模后每一周对小鼠的后足关节炎体征进行评分;采集所有小鼠关节滑膜细胞,采用PCR法检测细胞中RANKL、Smadl水平;采用Westernblot法检测细胞中碱性磷酸酶(ALP)、骨谷氨酸蛋白(BGP)、DKK1、Wnt5a水平。结果造模后模型组、阳性药物组、丹皮酚组小鼠关节炎症评分无显著差异(P>0.05),但均高于空白组(P<0.05);治疗1周、治疗2周及治疗3周时阳性药物组及丹皮酚组小鼠后足关节炎体征评分均明显低于模型组,差异有统计学意义(P<0.05);治疗后,模型组小鼠RANK、Smadl、Wnt5a水平均高于空白组,ALP、BGP、DKK1水平低于空白组(均P<0.05);阳性药物组、丹皮酚组小鼠RANK、Smadl、Wnt5a水平均明显低于模型组,滑膜细胞中ALP、BGP、DKK1水平明显高于模型组,差异均有统计学意义(均P<0.05);阳性药物组与丹皮酚组小鼠各指标比较无显著差异(P>0.05)。结论采用丹皮酚对AS小鼠治疗后可有效改善小鼠后足炎症评分,并可有效调节Wnt通路,降低滑膜组织中RANKL及Smadl水平。

Objective To analyze the therapeutic effect of Paeonol on ankylosing spondylitis(AS)mice and its effects on Wnt pathway,RANKL and Smalll levels.Methods 100 rats were randomly divided into model group,positive drug group,blank group and paeonol group.Except blank group,all the other three groups were modeled with 100μl proteoglycan+100μl Freund’s complete adjuvant.After successful modelling,the positive drug group was given 9 mg/kg sulfasalazine by gavage,the Paeonol group was given 3 mg/kg paeonol by gavage,and the model group and blank group were given equal volume distillation water.Each group was administered once a day for 20 consecutive days;the signs of hind foot arthritis in mice were scored before and every week after the establishment of the model;the synovial cells of all mice were collected and the levels of RANKL and Smadl in the cells were detected by PCR;the levels of ALP,BGP,DKK1 and Wnt5 a in the cells were detected by Western blot.Results There was no significant difference in the scores of arthritis in the model group,positive drug group and paeonol group after modeling(P>0.05),but all were higher than those in the blank group(P<0.05).After treatment,the levels of RANK,Smadl,and Wnt5 a in the model group were higher than those in the blank group,and the levels of ALP,BGP,and DKK1 were lower than those in the blank group(all P<0.05);The Wnt5 a level was significantly lower than the model group,and the levels of ALP,BGP,and DKK1 in synovial cells were significantly higher than the model group,and the differences were statistically significant(all P<0.05).There was no significant difference in ALP,BGP,DKK1 and Wnt5 a levels in membrane cells(P>0.05).Conclusion Paeonol can effectively improve the inflammation score of hind foot in AS mice,regulate Wnt pathway and reduce the levels of RANKL and Smadl in synovial tissue.