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1.
PLoS One ; 15(6): e0234747, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32584845

RESUMO

SUMMARY OF BACKGROUND DATA: The vascular buds in the vertebral endplate (VEP) are the structural foundation of nutrient exchange in the intervertebral disc (IVD). VEGF is closely related to angiogenesis in the endplate and intervertebral disc degeneration (IDD). OBJECTIVE: To investigate the effects of static load on vascular buds and VEGF expression in the VEP and to further clarify the relation between IDD and VEGF. METHODS: IVD motion segments were harvested from rabbit lumbar spines and cultured under no-loading conditions (controls) or in custom-made apparatuses under a constant compressive load (0.5 MPa) for up to 14 days. Tissue integrity and the number of vascular buds were determined, and the concentrations and expression of Aggrecan, COL2a1, and VEGFA in the VEPs were assessed after 3, 7, and 14 days of culturing and then compared with those of fresh tissues. RESULTS: Under the constant compression, the morphological integrity of the VEPs was gradually disrupted, and immunohistochemistry results showed a significant decrease in the levels of Agg and COL2a1. During the static load, the number of vascular buds in the VEPs was gradually reduced from the early stage of culture, and ELISA showed that the constant compressive load caused a significant decrease in the VEGFA and VEGFR2 protein concentrations, which were consistent with the immunohistochemistry results. Western blot and RT-PCR results also showed that the loading state caused a significant decrease in VEGFA expression compared with that of fresh and control samples. CONCLUSIONS: Constant compression caused degeneration of the VEP as well as a decreased number of vascular buds, thereby accelerating disc degeneration. VEGFA is involved in this process. We anticipate that regulating the expression of VEGFA may improve the condition of the lesions to the vascular buds in the endplates, thus enhancing the nutritional supply function in IVD and providing new therapeutic targets and strategies for the effective prevention and treatment of IDD.

2.
Zhongguo Zhong Yao Za Zhi ; 45(3): 609-616, 2020 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-32237520

RESUMO

The aim of this paper was to investigate the key targets and mechanism of "Epimedii Folium-Paeoniae Radix Alba" in the treatment of lumbar disc herniation by means of network pharmacology. The currently recognized databases and analysis software at home and abroad were used to construct the network from drugs and diseases. The chemical components of Epimedii Folium and Paeo-niae Radix Alba were collected by using databases such as TCMSP, while their active components were determined and the action targets were predicted according to threshold screening and literature reports. The genes for lumbar disc herniation were collected by using GeneCards, OMIM, and DisGeNET databases. The drug targets were mapped to disease targets, and protein interaction network analysis for key targets, GO function enrichment analysis and KEGG signaling pathway enrichment analysis were performed. Finally, 23 active components of Epimedium Folium and 13 active components of Paeoniae Radix Alba were determined, and a total of 624 drug targets were obtained. After standardization, 214 drug targets were obtained. In addition, 306, 2 and 5 related targets of lumbar disc herniation were collected from GeneCards, OMIM, and DisGeNET database, respectively, and a total of 293 disease targets were obtained after deduplication. After the mapping of drug target and disease target, 44 common targets were obtained. PPI protein interaction network analysis showed that IL-6, TNF, AKT1, MAPK1, and VEGFA may be the core targets for the treatment of lumbar disc herniation. GO enrichment analysis identified 56 items(P<0.05), among which biological processes mainly included immune response, apoptosis, etc.; cell components mainly included extracellular space, extracellular region, etc.; molecular functions mainly included cytokine activity, metallopeptidase activity and so on. Through KEGG pathway enrichment analysis, 91 signaling pathways related to inflammation, metabolism, and senescence were identified, mainly including IL-17 signaling pathway and TNF signaling pathway and so on. "Epimedii Folium-Paeoniae Radix Alba" showed the characteristics of multi-channel and multi-target for the treatment of lumbar disc herniation. This study preliminarily explored the key targets for its role and the biological processes and signaling pathways involved. It was found that it may play a therapeutic role by affecting inflammation and immune regulation, which laid the foundation for further experimental verification.

3.
Medicine (Baltimore) ; 98(49): e18135, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31804324

RESUMO

BACKGROUND: Degenerative lumbar spondylolisthesis (DLS) is one of the common orthopedic diseases which causes low back pain in patients, which seriously affects people's daily life and work. As a method of conservative treatment of this disease, manipulation is widely used in clinical practice. We will summarize the current published evidence of manipulation in the treatment of DLS, and evaluate the effectiveness and safety of manipulation through systematic review and meta-analysis, so as to provide more reliable evidence for future clinical practice. METHODS: We will conduct a comprehensive search of the following 9 databases until January 2019: PubMed, Embase, Cochrane Library, ClinicalTrials.gov, Web of Science, Chinese National Knowledge Infrastructure, Chinese Science and Technique Journals Database, Wan Fang Database, and Chinese Biomedical Database. The 2 researchers will independently search, screen, extract data, and evaluate the quality of the literatures. The primary outcomes include clinical effectiveness, Japanese Orthopaedic Association scores, and the secondary outcomes include visual analog scale scores, symptom scores, and adverse events. Bias risk tools provided by Cochrane Collaboration will be used for literature quality assessment, and RevMan 5.3 software will be used for meta-analysis. RESULTS: The results of this study will systematically evaluate the effectiveness and safety of manipulation intervention for people with DLS, especially in improving lumbar function scores and pain scores. CONCLUSION: The systematic review of this study will summarize the current published evidence of manipulation for the treatment of DLS, which can further guide the promotion and application of it. ETHICS AND DISSEMINATION: This study does not require ethical approval and the results will be published in a peer-reviewed journal. PROSPERO REGISTRATION NUMBER: CRD42019139933.


Assuntos
Vértebras Lombares , Manipulação da Coluna/métodos , Espondilolistese/terapia , Humanos , Manipulação da Coluna/efeitos adversos , Ensaios Clínicos Controlados Aleatórios como Assunto , Projetos de Pesquisa
4.
Zhongguo Gu Shang ; 31(7): 627-634, 2018 Jul 25.
Artigo em Chinês | MEDLINE | ID: mdl-30103586

RESUMO

OBJECTIVE: To compare effect of serum of Bushen Huoxue prescription on rabbit with intervertebral disc motion segments in vitro culture. METHODS: Twenty-four New Zealand white rabbits aged from 4 to 6 months and weighted from 2.5 to 3 kg were divided into medicated group and control group, 7 in each group. Rabbits were excuted under condition of asepsis, 28 spinal motion segments were taken out in each group, and segments were loaded in spinal motion segments in vitro and cultured in culture apparatus. Nutrient solution of medicated group contain 10% serum of Bushen Huoxue prescription, and 10% blank serum in control group. Seven discs between two groups were taken out and observed by histomorphology, proteoglycan PAS/AB stining, collagen II immunohistochemical staining, AGG, Col2aI by PCR test before culture and on the 3th, 7th and 14th day after culture. RESULTS: Histomorphology results showed the formation of medicated group was better than that of control group at 1 week after culture. PAB/AB test results showed content of poteoglycan between two groups were decreased. Imunohistochemical results showed content of collagen II in medicated group were obviously increased than that of control group at 3 days, but decreased obviously at 1 week than that of control group. PCR results showed expression of Agg in medicated group was obviously decreased than that of control group, but no statistical significance between two groups on the 7 th and 14 th day. Expression of Col2aI between two groups at 3 days were increased, and medicated group increased obviously more, while there were no significant difference between two groups. CONCLUSIONS: Serum of Bushen Huoxue prescription could delay intervertebral disc degeneration at short time for it is relate with improving circulation, inhibiting inflammatory, regulating extracellular matrix, so the prescription plays an important role in early prevention and treatment for intervertebral disc degeneration.


Assuntos
Degeneração do Disco Intervertebral , Disco Intervertebral , Animais , Medicamentos de Ervas Chinesas , Matriz Extracelular , Coelhos
5.
Biomed Res Int ; 2016: 2481712, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27872846

RESUMO

The development of mechanically active culture systems helps in understanding of the role of mechanical stress in intervertebral disc (IVD) degeneration. Motion segment cultures facilitate the application and control of mechanical loads. The purpose of this study was to establish a culturing method for rabbit IVD motion segments to observe the effect of static load on the whole disc organ. Segments were cultured in custom-made apparatuses under a constant, compressive load (3 kg) for 2 weeks. Tissue integrity, matrix synthesis, and matrix gene expression profile were assessed and compared with fresh one. The results showed ex vivo culturing of samples gradually destroyed the morphology. Proteoglycan contents and gene expression were decreased and downregulated obviously. However, immunohistochemical staining intensity and collagen type II gene expression were significantly enhanced and upregulated. In contrast, these trends were reversed under constant compression. These results indicated short-term static load stimulated the synthesis of type II collagen; however, constant compression led to progressive degeneration and specifically to proteoglycan. Through this study a loading and organ-culturing system for ex vivo rabbit IVD motion segments was developed, which can be used to study the effects of mechanical stimulation on the biology of IVDs and the pathomechanics of IVD degeneration.


Assuntos
Colágeno Tipo II/biossíntese , Matriz Extracelular/metabolismo , Regulação da Expressão Gênica , Degeneração do Disco Intervertebral/metabolismo , Disco Intervertebral/metabolismo , Estresse Mecânico , Animais , Modelos Animais de Doenças , Matriz Extracelular/patologia , Disco Intervertebral/patologia , Disco Intervertebral/fisiopatologia , Degeneração do Disco Intervertebral/patologia , Degeneração do Disco Intervertebral/fisiopatologia , Coelhos , Suporte de Carga
6.
Chin Med J (Engl) ; 129(19): 2338-46, 2016 10 05.
Artigo em Inglês | MEDLINE | ID: mdl-27647194

RESUMO

BACKGROUND: The development of mechanically active culture systems helps increase the understanding of the role of mechanical stress in intervertebral disc (IVD) degeneration. Motion segment cultures allow for preservation of the native IVD structure, and adjacent vertebral bodies facilitate the application and control of mechanical loads. The purpose of this study was to establish loading and organ culture methods for rabbit IVD motion segments to study the effect of static load on the whole disc organ. METHODS: IVD motion segments were harvested from rabbit lumbar spines and cultured in no-loading 6-well plates (control conditions) or custom-made apparatuses under a constant, compressive load (3 kg, 0.5 MPa) for up to 14 days. Tissue integrity, matrix synthesis, and the matrix gene expression profile were assessed after 3, 7, and 14 days of culturing and compared with those of fresh tissues. RESULTS: The results showed that ex vivo culturing of motion segments preserved tissue integrity under no-loading conditions for 14 days whereas the static load gradually destroyed the morphology after 3 days. Proteoglycan contents were decreased under both conditions, with a more obvious decrease under static load, and proteoglycan gene expression was also downregulated. However, under static load, immunohistochemical staining intensity and collagen Type II alpha 1 (COL2A1) gene expression were significantly enhanced (61.54 ± 5.91, P = 0.035) and upregulated (1.195 ± 0.040, P = 0.000), respectively, compared with those in the controls (P < 0.05). In contrast, under constant compression, these trends were reversed. Our initial results indicated that short-term static load stimulated the synthesis of collagen Type II alpha 1; however, sustained constant compression led to progressive degeneration and specifically to a decreased proteoglycan content. CONCLUSIONS: A loading and organ culture system for ex vivo rabbit IVD motion segments was developed. Using this system, we were able to study the effects of mechanical stimulation on the biology of IVDs, as well as the pathomechanics of IVD degeneration.


Assuntos
Disco Intervertebral/metabolismo , Disco Intervertebral/fisiologia , Núcleo Pulposo/metabolismo , Núcleo Pulposo/fisiologia , Técnicas de Cultura de Órgãos/métodos , Estresse Mecânico , Animais , Regulação da Expressão Gênica , Imuno-Histoquímica , Degeneração do Disco Intervertebral/metabolismo , Degeneração do Disco Intervertebral/fisiopatologia , Masculino , Coelhos
7.
Zhongguo Gu Shang ; 28(9): 824-31, 2015 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-26647565

RESUMO

OBJECTIVE: To compare the changes of nucleus pulposus after in vitro culture of rabbit whole intervertebral disc and spinal motion segment. METHODS: Twenty-one New Zealand white rabbits which were randomly divided into organ group with 8 rabbits and segment group with 13 rabbits. Fifty intervertebral discs and 50 spinal motion segments were harvested respectively under aseptic conditions from two groups. These specimens were maintained in organ culture with hyperosmotic media (410 mOsm/kg), then 10 discs of the two groups were observed respectively by HE staining, immunohistochemistry of collagen type III, proteoglycan content and cells viability of nucleus pulposus before culture and at 3, 7, 14, 21 days after culture. RESULTS: HE staining showed the intervertebral disc tissue structure remained intact after culture of 21 days organ group and 14 days segment group,but there was severely degenerated of 21 days segment group. The intensity value of type II collagen immunohistochemical staining in the nucleus pulposus were not changed significantly between 21 days organ group and 14 days segment group (P > 0.05), but the staining of segment group at 21 days became shallower, there was significant difference compared with before each time points and organ group at 21 days (P < 0.05). PAS/AB staining of proteoglycan of nucleus pulposus showed that there were not decrease of tinting strength of two groups within 7 days, but the strength weakened slightly of two groups at 14 days, and the tinting strength became weaker at 21 days segment group, the change is more obvious than the organ group. The intensity value of fluorescence staining of nucleus pulposus cells was not changed significantly within 7 days of two groups (P > 0.05), the intensity value decreased slightly at 21 days organ group and 14 days segment group, but there were no significant difference compared with before time points (P > 0.05) however at 21 days segment group the intensity decreased as cells viability of nucleus pulposus decreased,and there was a significant difference compared with before each time points and organ group at 21 days (P < 0.05). CONCLUSION: It is not obviously degenerated of the discs of organ group cultured within 21 days and segment group cultured within 14 days, but there was significant degeneration of the intervertebral disc of segment group after cultured 21 days, so the rabbit spinal motion segment can be used on research about the biomechanics of intervertebral disc as a vitro experimental model within 14 days.


Assuntos
Disco Intervertebral/patologia , Animais , Colágeno Tipo II/análise , Feminino , Imuno-Histoquímica , Disco Intervertebral/química , Masculino , Técnicas de Cultura de Órgãos , Coelhos
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