Mitochondrial dependent pathway is involved in the protective effects of carboxymethylated chitosan on nitric oxide-induced apoptosis in chondrocytes
Abstract Background Chondrocyte apoptosis activated by the mitochondrial dependent pathway serves a crucial role in cartilage degeneration of osteoarthritis (OA). In the present study, the protective effects of CMCS against sodium nitroprusside (SNP)-induced chondrocyte apoptosis were evaluated and...
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BMC
2020-01-01
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| Series: | BMC Complementary Medicine and Therapies |
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| Online Access: | https://doi.org/10.1186/s12906-019-2808-x |
| _version_ | 1818916336664838144 |
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| author | Bin He Fei Wu Xiaohai Li Yang Liu Li Fan Haohuan Li |
| author_facet | Bin He Fei Wu Xiaohai Li Yang Liu Li Fan Haohuan Li |
| author_sort | Bin He |
| collection | DOAJ |
| description | Abstract Background Chondrocyte apoptosis activated by the mitochondrial dependent pathway serves a crucial role in cartilage degeneration of osteoarthritis (OA). In the present study, the protective effects of CMCS against sodium nitroprusside (SNP)-induced chondrocyte apoptosis were evaluated and the underlying molecular mechanisms were elucidated. Methods Chondrocytes were isolated from articular cartilage of SD rats and identified by type II collagen immunohistochemistry. The chondrocytes stimulated with or without SNP to induce apoptosis, were treated by CMCS for various concentrations. The cell viability were determined by MTT and LDH assays. Cell apoptotic ratio was determined by Annexin V-FITC/PI staining. Mitochondrial membrane potential (ΔΨm) was detected by using Rhodamine123 (Rho123) staining. To understand the mechanism, the mRNA expression levels of Bcl-2, Bax, cytochrome c (Cyt c) and cleaved caspase-3 were detected by real-time PCR and western blot analysis, respectively. Results It was shown using the MTT and LDH assays that CMCS protected the viability of chondrocyte against SNP damage. Annexin V-FITC/PI and Rho123 staining showed that CMCS not only inhibited the cell apoptosis but also restored the reduction of the ΔΨm in chondrocytes. In SNP-induced chondrocytes, CMCS down-regulated the expression of Bax, Cyt c and cleaved caspase-3 but upregulated the expression of Bcl-2, as shown by real-time PCR and western blot. Conclusions Taken together, these results indicated that CMCS has the protective effect on chondrocytes against SNP-induced apoptosis, at least partly, via inhibiting the mitochondrial dependent apoptotic pathway. Thus, CMCS may be potentially used as a biological agent for prevention and treatment of OA. |
| first_indexed | 2024-12-20T00:16:33Z |
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| institution | Directory Open Access Journal |
| issn | 2662-7671 |
| language | English |
| last_indexed | 2024-12-20T00:16:33Z |
| publishDate | 2020-01-01 |
| publisher | BMC |
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| series | BMC Complementary Medicine and Therapies |
| spelling | doaj.art-c795f232297b4c42853b17d1c449bbc22022-12-21T20:00:21ZengBMCBMC Complementary Medicine and Therapies2662-76712020-01-0120111010.1186/s12906-019-2808-xMitochondrial dependent pathway is involved in the protective effects of carboxymethylated chitosan on nitric oxide-induced apoptosis in chondrocytesBin He0Fei Wu1Xiaohai Li2Yang Liu3Li Fan4Haohuan Li5Department of Orthopaedics, Renmin Hospital of Wuhan UniversityDepartment of Orthopaedics, Renmin Hospital of Wuhan UniversityDepartment of Orthopaedics, Renmin Hospital of Wuhan UniversityDepartment of Orthopaedics, Renmin Hospital of Wuhan UniversityDepartment of Orthopaedics, Renmin Hospital of Wuhan UniversityDepartment of Orthopaedics, Renmin Hospital of Wuhan UniversityAbstract Background Chondrocyte apoptosis activated by the mitochondrial dependent pathway serves a crucial role in cartilage degeneration of osteoarthritis (OA). In the present study, the protective effects of CMCS against sodium nitroprusside (SNP)-induced chondrocyte apoptosis were evaluated and the underlying molecular mechanisms were elucidated. Methods Chondrocytes were isolated from articular cartilage of SD rats and identified by type II collagen immunohistochemistry. The chondrocytes stimulated with or without SNP to induce apoptosis, were treated by CMCS for various concentrations. The cell viability were determined by MTT and LDH assays. Cell apoptotic ratio was determined by Annexin V-FITC/PI staining. Mitochondrial membrane potential (ΔΨm) was detected by using Rhodamine123 (Rho123) staining. To understand the mechanism, the mRNA expression levels of Bcl-2, Bax, cytochrome c (Cyt c) and cleaved caspase-3 were detected by real-time PCR and western blot analysis, respectively. Results It was shown using the MTT and LDH assays that CMCS protected the viability of chondrocyte against SNP damage. Annexin V-FITC/PI and Rho123 staining showed that CMCS not only inhibited the cell apoptosis but also restored the reduction of the ΔΨm in chondrocytes. In SNP-induced chondrocytes, CMCS down-regulated the expression of Bax, Cyt c and cleaved caspase-3 but upregulated the expression of Bcl-2, as shown by real-time PCR and western blot. Conclusions Taken together, these results indicated that CMCS has the protective effect on chondrocytes against SNP-induced apoptosis, at least partly, via inhibiting the mitochondrial dependent apoptotic pathway. Thus, CMCS may be potentially used as a biological agent for prevention and treatment of OA.https://doi.org/10.1186/s12906-019-2808-xCarboxymethylated chitosanOsteoarthritisChondrocytesApoptosis |
| spellingShingle | Bin He Fei Wu Xiaohai Li Yang Liu Li Fan Haohuan Li Mitochondrial dependent pathway is involved in the protective effects of carboxymethylated chitosan on nitric oxide-induced apoptosis in chondrocytes BMC Complementary Medicine and Therapies Carboxymethylated chitosan Osteoarthritis Chondrocytes Apoptosis |
| title | Mitochondrial dependent pathway is involved in the protective effects of carboxymethylated chitosan on nitric oxide-induced apoptosis in chondrocytes |
| title_full | Mitochondrial dependent pathway is involved in the protective effects of carboxymethylated chitosan on nitric oxide-induced apoptosis in chondrocytes |
| title_fullStr | Mitochondrial dependent pathway is involved in the protective effects of carboxymethylated chitosan on nitric oxide-induced apoptosis in chondrocytes |
| title_full_unstemmed | Mitochondrial dependent pathway is involved in the protective effects of carboxymethylated chitosan on nitric oxide-induced apoptosis in chondrocytes |
| title_short | Mitochondrial dependent pathway is involved in the protective effects of carboxymethylated chitosan on nitric oxide-induced apoptosis in chondrocytes |
| title_sort | mitochondrial dependent pathway is involved in the protective effects of carboxymethylated chitosan on nitric oxide induced apoptosis in chondrocytes |
| topic | Carboxymethylated chitosan Osteoarthritis Chondrocytes Apoptosis |
| url | https://doi.org/10.1186/s12906-019-2808-x |
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