基于病理表型辨析激素性与酒精性股骨头坏死的血瘀证特点

Study on characteristics of differentiation of blood stasis syndrome between steroid and alcohol induced osteonecrosis of the femoral head based on pathological phenotype

目的:观察激素性和酒精性股骨头坏死(ONFH)患者骨组织中VEGF、β-catenin、OPG、RANKL蛋白的差异表达,探讨二者血瘀证发展的特点。方法:随机选取2017年1月至2018年12月因激素性和酒精性ONFH行人工全髋关节置换术的股骨头标本各30例(激素组、酒精组),及对应患者的血清样本。应用Western Blot分别检测股骨头内坏死区、硬化区和正常区的骨组织中VEGF、β-catenin、OPG、RANKL蛋白表达水平;ELISA检测患者血清中对应的蛋白含量。结果:两组坏死区与硬化区间有死骨吸收。激素组吸收囊变区可见带血运的肉芽组织,酒精组吸收区内填充纤维组织,血运新生不明显;激素组各区骨组织VEGF、RANKL蛋白表达量、RANKL/OPG比值均高于酒精组(P<0.01),硬化区和正常区OPG蛋白表达量低于酒精组(P<0.05),坏死区和正常区β-catenin蛋白表达量低于酒精组(P<0.05),OPG/RANKL比值均低于酒精组(P<0.01)。激素组血清VEGF、RANKL浓度及RANKL/OPG比值高于酒精组(P<0.05),β-catenin、OPG浓度及OPG/RANKL比值低于酒精组(P<0.05)。结论:激素组骨吸收与血管化能力强,血瘀证可能与应用激素使毒邪内蕴,耗气伤阴,阻滞气机,运血无力有关;酒精组骨吸收与血管化能力弱,血瘀证可能与饮酒损伤脾胃,脾胃为气血生化之源,气血生化乏源,气血两虚有关。

Objective:To observe the differential expression of VEGF,β-catenin,OPG and RANKL protein in bone tissue of patients with steroid-induced osteonecrosis of the femoral head(SONFH)and alcohol-induced osteonecrosis of the femoral head(AONFH),and further analyze the characteristics of the development of blood stasis syndrome.Methods:A total of 60 femoral head specimens and serum samples of corresponding patients were randomly selected for total hip arthroplasty due to SONFH and AONFH from January 2017 to December 2018.Western Blot was used to detect the expression of VEGF,β-catenin,OPG and RANKL in necrosis area,sclerotic area and normal area of femoral head,and ELISA was used to detect the protein concentration of VEGF,β-catenin,OPG and RANKL in serum.Results:Two groups necrosis area and sclerotic area had dead bone absorption.In the steroid group,granulation tissue with learning transport could be seen in the absorption zone,while in the alcohol group,fibrous tissue was filled in the absorption zone,but the blood transport was not obvious.The expression of VEGF and RANKL protein,the ratio of RANKL/OPG in osteonecrosis area,sclerotic area and normal area of steroid group was higher than that of alcohol group(P<0.01),the expression of OPG protein in sclerotic area and normal area was lower than that of alcohol group(P<0.05),the expression ofβ-catenin protein in necrosis area and normal area was lower than that of alcohol group(P<0.05),the ratio of RANKL/OPG in each area was lower than that of alcohol group(P<0.01).The serum VEGF,RANKL concentration and RANKL/OPG ratio in steroid group were higher than those in alcohol group(P<0.05),whileβ-catenin,OPG concentration and OPG/RANKL ratio in steroid group were lower than those in alcohol group(P<0.05).Conclusion:The ability of bone absorption and vascularization is strong in steroid group,and the formation of blood stasis syndrome may be related to the application of hormone to make poison and evil accumulate,the consumption of qi to injure yin,the block of qi mechanism,and the weakness of blood circulation;the ability of bone absorption and vascularization is weak in alcohol group,and the formation of blood stasis syndrome may be related to the damage of spleen and stomach by drinking alcohol,which is the source of blood biochemistry,the lack of blood biochemistry and the deficiency of both qi and blood.