■探讨中药灵芝(LZ)、三苗散(SMS)联合透明质酸(HA)凝胶减轻创伤性骨关节炎(OA)软骨退变的疗效。
■前交叉韧带横切术(ACLT)的标准化手术是从8周龄雌性SD大鼠的右后肢内侧室进行的,并导致创伤性OA。大鼠(n=5/组)接受了一次关节内注射50μlHA凝胶治疗,50μlHA-凝胶+50μgLZ-SMS,50μl盐水+50μgLZ-SMS和null(ACLT组),假组除外。治疗后3个月收获四肢进行μCT扫描和组织病理学染色。使用免疫学多重分析试剂盒检测来自血浆和滑液的炎性细胞因子。将LZ-SMS中活性化合物的假定靶标与OA的已知治疗靶标组合以构建蛋白质-蛋白质相互作用网络。通过DAVID生物信息学工具,采用基因本体论和京都基因和基因组百科全书(KEGG)富集分析来预测LZ-SMS在OA中的潜在靶标和信号通路。
■体内,HA-gel+LZ-SMS治疗导致透明软骨(HC)/钙化软骨(CC)和HC/Sum(软骨总体积)的体积比更高,与ACLT和HA凝胶组相比。此外,组织学结果显示软骨基质升高,HA-gel+LZ-SMS治疗中的软骨形成和成骨细胞信号。治疗还显著改变了软骨下骨(SCB)结构,包括BV/TV的增加,TB。Th,BMD,康恩Dn,TB。N,还有DA,以及Tb的显着降低。Sp和Po(TOT),这暗示了对维持胫骨SCB微结构稳定的保护作用。此外,HA+LZ-SMS组炎症细胞因子下调,抗炎细胞因子IL-10上调.最后,来自LZ-SMS中37种活性化合物的64个共享靶标与OA发展的核心基因相关。LZ-SMS通过影响MAPK信号通路在调节炎症环境中具有推定的作用。
■我们的研究阐明了HA-gel+LZ-SMS在减轻软骨降解以及与外伤性OA发展的靶标和信号通路的假定相互作用方面的保护作用。
■我们的结果为使用LZ-SMS作为OA治疗的潜在候选者提供了生物学原理。
UNASSIGNED: To investigate the mitigate efficacy of Chinese medicine Lingzhi (LZ) and San-Miao-San (SMS) combined with hyaluronic acid (HA)-gel in attenuating cartilage degeneration in traumatic osteoarthritis (OA).
UNASSIGNED: The standardized surgery of anterior cruciate ligament transection (ACLT) was made from the medial compartment of right hind limbs of 8-week-old female SD rats and resulted in a traumatic OA. Rats (n = 5/group) were treated once intra-articular injection of 50 μl HA-gel, 50 μl HA-gel+50 μg LZ-SMS, 50 μl of saline+50 μg LZ-SMS and null (ACLT group) respectively, except sham group. Limbs were harvested for μCT scan and histopathological staining 3-month post-treatment. Inflammatory cytokines from plasma and synovial fluid were detected using Immunology Multiplex Assay kit. The putative targets of active compounds in LZ-SMS and known therapeutic targets for OA were combined to construct protein-protein interaction network. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was adopted to predict the potential targets and signaling pathway of LZ-SMS in OA through the tool of DAVID Bioinformatics.
UNASSIGNED: In vivo, HA-gel + LZ-SMS treatment resulted in a higher volume ratio of hyaline cartilage (HC)/calcified cartilage (CC) and HC/Sum (total volume of cartilage), compared to ACLT and HA-gel groups. In addition, histological results showed the elevated cartilage matrix, chondrogenic and osteoblastic signals in HA-gel + LZ-SMS treatment. Treatment also significantly altered subchondral bone (SCB) structure including an increase in BV/TV, Tb.Th, BMD, Conn.Dn, Tb.N, and DA, as well as a significant decrease in Tb.Sp and Po(tot), which implied a protective effect on maintaining the stabilization of tibial SCB microstructure. Furthermore, there was also a down-regulated inflammatory cytokines and upregulated anti-inflammatory cytokine IL-10 in HA+LZ-SMS group. Finally, 64 shared targets from 37 active compounds in LZ-SMS related to the core genes for the development of OA. LZ-SMS has a putative role in regulating inflammatory circumstance through influencing the MAPK signaling pathway.
UNASSIGNED: Our study elucidated a protective effect of HA-gel + LZ-SMS in mitigating cartilage degradation and putative interaction with targets and signaling pathway for the development of traumatic OA.
UNASSIGNED: Our results provide a biological rationale for the use of LZ-SMS as a potential candidate for OA treatment.