探讨白芍总苷胶囊对聚乙烯颗粒介导的小鼠颅骨骨溶解的作用

Effect of Total Glucosides of Paeony Capsules on Osteolysis of Mouse Skull Mediated by Polyethylene Particles

目的探讨白芍总苷胶囊对超高分子聚乙烯颗粒介导的小鼠颅骨骨溶解的作用及机制。方法将60只8周龄的C57BL/6J小鼠随机分为5组:假手术组(Sham组)、模型对照组(Vehicle组)、白芍总苷低剂量组(LTGP组)、白芍总苷中剂量组(M-TGP组)、白芍总苷高剂量组(H-TGP组)。造模成功后,Sham组、Vehicle组用等体积生理盐水连续灌胃2周,L-TGP、M-TGP、H-TGP组分别用含低(0.1175 g·kg^-1)、中(0.235 g·kg^-1)、高(0.47 g·kg^-1)剂量的白芍总苷药液连续灌胃2周。14 d后采集小鼠颅骨进行显微计算机断层扫描(micro CT),记录骨体积分数(BV/TV),并进行统计学分析;应用苏木精-伊红染色法(hematoxylin-eosin staining,HE)进行组织学分析;应用免疫组化技术(immunohistochemistry,IHC)检测小鼠颅骨中的白介素1(interleukin 1,IL-1)、白介素6(interleukin 6,IL-6)、白细胞分化抗原68(cluster of differentiation 68,CD68)等炎症因子的表达。结果Micro CT的结果提示,相对于Sham组,Vehicle组小鼠颅骨的BV/TV值降低,差异具有统计学意义(P<0.01),Vehicle组小鼠颅骨有明显的骨溶解现象;L-TGP、M-TGP、H-TGP组的BV/TV值均高于Vehicle组(P<0.05,P<0.01),M-TGP组的BV/TV值较L-TGP组、H-TGP组高,M-TGP组与L-TGP组的差异具有统计学意义(P<0.01),H-TGP组与L-TGP组差异无统计学意义(P>0.05)。HE染色结果提示Vehicle组颅骨骨吸收明显,骨小梁稀疏且有断裂现象,细胞形态不规则。M-TGP组颅骨骨小梁排列整齐,骨小梁未见明显断裂,炎症细胞较L-TGP、H-TGP组少。免疫组化结果提示TGP可以通过抑制CD68、IL-1、IL-6的表达,从而对破骨细胞的生成有一定的抑制作用。其中,M-TGP与L-TGP、H-TGP相比,对炎症因子的抑制作用较明显。结论白芍总苷胶囊可抑制炎症反应,抑制小鼠颅骨骨溶解,可能是治疗由炎症因子高表达引起的溶骨性疾病的有效药物,但其有效性尚需更深入的实验进行验证。

Objective To investigate the effect and mechanism of total glucosides of paeony capsules on ultrahighmolecular polyethylene(UHMPE)-mediated osteolysis of mouse skull.Methods Sixty C57BL/6J mice(aged 8 weeks)were randomly divided into 5 groups:Sham group,Vehicle group,low dose group(L-TGP group),medium dose group(M-TGP group),and high dose group(H-TGP group).After modeling,the Sham group and the Vehicle group were given normal saline for two consecutive weeks,and the L-TGP,M-TGP,H-TGP groups were given 0.2 mL of total glucosides of paeony capsules at low(0.1175 g·kg^-1),medium(0.235 g·kg^-1)and high(0.47 g·kg^-1)doses respectively for two consecutive weeks.After 14 days,the mice’s skulls were collected for Micro computed tomography(Micro CT),the bone volume fraction(BV/TV)were recorded and statistical analysis was performed.Hematoxylin-eosin staining(HE)was used to analyze the mice’s skulls,and immunohistochemistry(IHC)was also used to detect the expressions of inflammatory factors such as Interleukin 1(IL-1),Interleukin 6(IL-6),and cluster of differentiation 68(CD68)in the skulls of mice.Results Compared with Sham group,the BV/TV value in Vehicle group was significantly decreased(P<0.05).The skulls of mice in Vehicle group showed significant osteolysis,and the BV/TV values of L-TGP,M-TGP and H-TGP groups were all higher than that of Vehicle group(P<0.05),and the BV/TV value of M-TGP was the highest.The difference of BV/TV between M-TGP group and L-TGP group was statistically significant(P<0.01),while the difference of BV/TV between H-TGP group and LTGP group was not statistically significant(P>0.05).The results of HE staining indicated that the skull bone absorption was obvious in the Vehicle group,the trabeculae were sparse and fractured,and the cell morphology was irregular.The trabeculae of the skull in the M-TGP group were orderly arranged,with no obvious fracture.There were fewer inflammatory cells in the M-TGP group than in the L-TGP group and the H-TGP group.The results of IHC suggested that TGP could inhibit the expression of CD68,Il-1 and Il-6,and medium dose of TGP showed a more obvious inhibitory effect on inflammatory factors than low and high doses.Conclusion Total glucosides of paeony capsules could inhibit the inflammatory reaction and the osteolysis of the skull of mice,which may be an effective drug for the treatment of osteolytic diseases caused by the high expression of inflammatory factors,but its effectiveness needs further evidence.