The Chinese Medicinal Formulation Guzhi Zengsheng Zhitongwan Modulates Chondrocyte Structure, Dynamics, and Metabolism by Controlling Multiple Functional Proteins

Yao, Baojin; Lu, Bocheng; Zhang, Mei; Gao, Hongwei; Leng, Xiangyang; Zhao, Daqing

doi:10.1155/2018/9847286

Cited by 4 publications

(11 citation statements)

References 52 publications

(50 reference statements)

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“…The tagged samples were pooled together and dried by vacuum centrifugation. The iTRAQ-labeled peptide mixtures were reconstituted with 4 mL strong cation exchange (SCX) buffer A (25 mM NaH 2 PO 4 in 25% acetonitrile (ACN), pH 2.7) and loaded onto a LC-20AB high-performance liquid chromatography platform (Shimadzu, Japan) equipped with a 4.6 × 250 mm Ultremex strong cation exchange column containing 5-μm particles (Phenomenex, USA) [2].…”

Section: Protein Digestion Itraq Labeling and Peptide Fractionationmentioning

confidence: 99%

“…Each fraction was further subjected to a TripleTOF 5600 mass spectrometer system (AB SCIEX, USA) equipped with a nanoACQuity UPLC system (Waters, USA) according to previously described [2]. Briefly, the peptide sample from each SCX fraction was loaded onto a nano-ACQuity UPLC BEH130 column (Waters, USA) packed with Symmetry C18 resin (Waters, USA).…”

Section: Nano-hplc-ms/ms Analysismentioning

confidence: 99%

“…The Mascot (Matrix Sciences, UK) and Proteome Discoverer software (Thermo, USA) were used for protein identification as previously described [2]. The collected raw data were converted into MGF files using the Proteome Discoverer software.…”

Section: Data Processingmentioning

confidence: 99%

“…The protein expression levels obtained by the iTRAQ analysis were further validated by the parallel reaction monitoring (PRM) assays as previously described [2]. Briefly, the spectral library was generated by the SpectroDive software (Biognosys AG, Switzerland), and the candidate proteins were obtained from the Data Dependent Acquisition (DDA) analysis mode that were generated by a Q-EXACTIVE mass spectrometer system (Thermo, USA).…”

Section: Parallel Reaction Monitoring Validationmentioning

confidence: 99%

“…(baked) and Cibotium barometz (L.) (J.Sm), is an effective formula of traditional Chinese herbal medicine and has been widely applied in the treatment of joint diseases for many years. Our previous studies have shown that the effects of GZZSZTW may be partially achieved via controlling chondrocyte proliferation and differentiation by modulating genes and proteins involved in chondrocyte structure, dynamics, and metabolism [1,2]. However, the precise molecular mechanism underlying its efficacy in treating joint diseases remains to be elucidated.…”

Section: Introductionmentioning

confidence: 99%

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Dissection of the molecular targets and signaling pathways of Guzhi Zengsheng Zhitongwan based on the analysis of serum proteomics

Yao

Liu

et al. 2019

Chin Med

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Background: Guzhi Zengsheng Zhitongwan (GZZSZTW) is an effective formula of traditional Chinese herbal medicine and has been widely applied in the treatment of joint diseases for many years. The aim of this study was to dissect the molecular targets and signaling pathways of Guzhi Zengsheng Zhitongwan based on the analysis of serum proteomics. Methods: The Chinese herbs of GZZSZTW were immersed in 5 l distilled water and boiled with reflux extraction method. The extract was filtered, concentrated and freeze-dried. The chemical profile of GZZSZTW extract was determined by high-performance lipid chromatography (HPLC). The 7-week old Sprague-Dawley (SD) rats in GZZSZTW groups were received oral administration at doses of 0.8, 1.05, and 1.3 g/kg per day and the rats in blank group were fed with drinking water. Serum samples were collected from the jugular veins. Primary chondrocyte viability was evaluated by CCK-8 assay. A full spectrum of the molecular targets and signaling pathways of GZZSZTW were investigated by isobaric tags for relative and absolute quantitation (iTRAQ) analysis and a systematic bioinformatics analysis accompanied with parallel reaction monitoring (PRM) and siRNA validation. Results: GZZSZTW regulated a series of functional proteins and signaling pathways responsible for cartilage development, growth and repair. Functional classification analysis indicated that these proteins were mainly involved in the process of cell surface dynamics. Pathway analysis mapped these proteins into several signalling pathways involved in chondrogenesis, chondrocyte proliferation and differentiation, and cartilage repair, including hippo signaling pathway, cGMP-PKG signaling pathway, cell cycle and calcium signaling pathway. Protein-protein interaction analysis and siRNA knockdown assay identified an interaction network consisting of TGFB1,

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Section: Protein Digestion Itraq Labeling and Peptide Fractionationmentioning

confidence: 99%

Section: Nano-hplc-ms/ms Analysismentioning

confidence: 99%

Section: Data Processingmentioning

confidence: 99%

Section: Parallel Reaction Monitoring Validationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Dissection of the molecular targets and signaling pathways of Guzhi Zengsheng Zhitongwan based on the analysis of serum proteomics

Yao

Liu

et al. 2019

Chin Med

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Integrated RNA-Seq Analysis Uncovers the Potential Mechanism of the “Kidney Governing Bones” Theory of TCM

Yang

Wang

Zhang

et al. 2022

Evidence-Based Complementary and Alternative Medicine

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Background. As in philosophy of traditional Chinese medicine (TCM), the theory of “kidney governing bones” has been demonstrated by a series of scientific studies. Furthermore, many groups including ours have explored the molecular mechanisms related to bone development, growth, and regeneration using modern biology technologies, such as RNA sequencing (RNA-Seq) and isobaric tags for relative and absolute quantification (ITRAQ), and have demonstrated that the underlying molecular mechanisms were highly consistent with the “kidney governing bones” theory. Objective. Kidney-yang deficiency (YD), as a pathological condition, has a passive effect on the skeleton growth; more specifically, it is a state of skeletal metabolic disorder. However, the exact molecular mechanisms related to the “kidney governing bones” theory under the control of multiple organs and systems are still unknown. Methods. In this study, we performed RNA-Seq analysis to investigate and compare the gene expression patterns of six types of tissue (bone, cartilage, kidney, testicle, thyroid gland, and adrenal gland) from YD rats and normal rats and analyzed the interaction effects controlled by multiple functional genes and signaling pathways between those tissues. Results. Our results showed that, in the state of YD, the functions of bone and cartilage were inhibited. Furthermore, multiple organs involving the reproductive, endocrine, and urinary systems were also investigated, and our results showed that YD could cause dysfunctions of these systems by downregulating multiple functional genes and signaling pathways that positively regulate the homeostasis of these tissues. Conclusion. We ensure that “kidney governing bones” was not a simple change in a single gene but the changes in complex biological networks caused by functional changes in multiple genes. This also coincides with the holistic view of TCM, which holds that the human body itself is an organic whole and the functional activities of each organ coordinate with each other.

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Deciphering the potential pharmaceutical mechanism of Guzhi Zengsheng Zhitongwan on rat bone and kidney based on the “kidney governing bone” theory

et al. 2020

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Background: Guzhi Zengsheng Zhitongwan (GZZSZTW) is an effective Chinese medicinal formulation for the treatment of osteoarthritis (OA) designed according to the "kidney governing bone" theory, which has been widely used as a golden guide for treating bone and cartilage diseases in traditional Chinese medicine. The aim of this study was to explore the molecular mechanism underlying its effects on the bone and kidney. Methods: Preparation and quality control were performed as previously described. Since GZZSZTW is orally administered in the form of pills prepared in boiled water, the Chinese materia medica (CMM) mixture of this formula was extracted with distilled water by a reflux method and was then filtered through a 0.45-μm Hollow Fiber Cartridge (GE Healthcare, USA). The filtrate was freeze-dried by a Heto PowerDry LL3000 Freeze Dryer (Thermo, USA) and stored at − 80°C. The effects of GZZSZTW on gene expression and regulation of both kidney and bone tissues were investigated using a state-of-the-art RNA-seq technology. Results: We demonstrated that GZZSZTW could enhance kidney function and suppress bone formation and resorption by modulating the activities of osteoblast and osteoclast, and might subsequently contribute to the inhibition of osteophyte formation during the process of OA. These effects might be achieved by the synergistic interactions of various herbs and their active components in GZZSZTW, which increased the expression levels of functional genes participating in kidney function, regulation, and repair, and then decreased the expression levels of genes involved in bone formation and resorption. Thus, our findings were consistent with the "kidney governing bone" theory, which has been widely used as a guide in clinical practice for thousands of years. Conclusions: This study has deepened the current knowledge about the molecular effects of GZZSZTW on bone and kidney regulation. Furthermore, this study might be able to provide possible strategies to further prevent and treat joint diseases by using traditional Chinese medicinal formulations following the "kidney governing bone" theory.

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The Chinese Medicinal Formulation Guzhi Zengsheng Zhitongwan Modulates Chondrocyte Structure, Dynamics, and Metabolism by Controlling Multiple Functional Proteins

Cited by 4 publications

References 52 publications

Dissection of the molecular targets and signaling pathways of Guzhi Zengsheng Zhitongwan based on the analysis of serum proteomics

Dissection of the molecular targets and signaling pathways of Guzhi Zengsheng Zhitongwan based on the analysis of serum proteomics

Integrated RNA-Seq Analysis Uncovers the Potential Mechanism of the “Kidney Governing Bones” Theory of TCM

Deciphering the potential pharmaceutical mechanism of Guzhi Zengsheng Zhitongwan on rat bone and kidney based on the “kidney governing bone” theory

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