Chu Man, Wu Ping, Hong Ming, Zeng Huasong, Wong Chun Kwok, Feng Yu, Cai Zhe, Lu William Weijia
Faulty of Medical Technology, Shaanxi University of Chinese Medicine, Xianyang, China.
Department of Allergy, Immunology and Rheumatology, Guangzhou Women and Children's Medical Center, Guangzhou, Guangdong, China.
J Orthop Translat. 2020 Oct 21;26:132-140. doi: 10.1016/j.jot.2020.07.008. eCollection 2021 Jan.
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).
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.
, 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.
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.
Our results provide a biological rationale for the use of LZ-SMS as a potential candidate for OA treatment.
探讨中药灵芝（LZ）和三妙散（SMS）联合透明质酸（HA）凝胶对减轻创伤性骨关节炎（OA）软骨退变的疗效。
对8周龄雌性SD大鼠右后肢内侧进行标准化的前交叉韧带切断术（ACLT），造成创伤性OA。除假手术组外，将大鼠（每组n = 5）分别关节腔内注射50 μl HA凝胶、50 μl HA凝胶+50 μg LZ - SMS、50 μl生理盐水+50 μg LZ - SMS及不做处理（ACLT组）。治疗3个月后采集肢体进行μCT扫描和组织病理学染色。使用免疫多重分析试剂盒检测血浆和滑液中的炎性细胞因子。将LZ - SMS中活性成分的假定靶点与OA已知治疗靶点相结合，构建蛋白质 - 蛋白质相互作用网络。采用基因本体论（Gene Ontology）和京都基因与基因组百科全书（KEGG）富集分析，通过DAVID生物信息学工具预测LZ - SMS在OA中的潜在靶点和信号通路。
与ACLT组和HA凝胶组相比，HA凝胶+LZ - SMS治疗使透明软骨（HC）/钙化软骨（CC）以及HC/总和（软骨总体积）的体积比更高。此外，组织学结果显示HA凝胶+LZ - SMS治疗中软骨基质、软骨生成和成骨信号升高。治疗还显著改变了软骨下骨（SCB）结构，包括骨体积分数（BV/TV）、骨小梁厚度（Tb.Th）、骨密度（BMD）、骨小梁连接密度（Conn.Dn）、骨小梁数量（Tb.N）和骨小梁间距（DA）增加，以及骨小梁分离度（Tb.Sp）和总体孔隙率（Po(tot)）显著降低，这意味着对维持胫骨SCB微观结构的稳定性具有保护作用。此外，HA + LZ - SMS组炎性细胞因子下调，抗炎细胞因子白细胞介素 - 10上调。最后，LZ - SMS中37种活性成分的64个共享靶点与OA发展的核心基因相关。LZ - SMS可能通过影响丝裂原活化蛋白激酶（MAPK）信号通路在调节炎症环境中发挥作用。
我们的研究阐明了HA凝胶+LZ - SMS在减轻软骨退变方面的保护作用，以及与创伤性OA发展相关靶点和信号通路的假定相互作用。
我们的结果为使用LZ - SMS作为OA治疗的潜在候选药物提供了生物学依据。
