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| 壮筋养血汤介导MAPK/P38-ERK通路调控成骨成脂分化的作用机制 |
| The mechanism of Zhuangjin Yangxue Decoction regulating osteogenic and adipogenic differentiation through the MAPK/P38-ERK pathway |
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| DOI：10.3969/j.issn.1006-7108.2023.08.003 |
| 中文关键词: 壮筋养血汤 LncRNA ANRIL MAPK/ P38-ERK信号通路 成骨成脂分化 |
| 英文关键词:Zhuangjin Yangxue decoction LncRNA ANRIL MAPK/P38-ERK osteogenic and adipogenic differentiation |
| 基金项目:国家自然科学基金面上项目(82272526) |
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| 中文摘要: |
| 目的 探讨壮筋养血汤（Zhuangjin Yangxue Decoction，ZJYXD）通过LncRNA ANRIL介导的MAPK/ P38-ERK 通路调控成骨成脂分化的作用机制。方法 提取骨髓间充质干细胞（BMSCs），并采用ANRIL慢病毒转染BMSCs；制备ZJYXD含药血清用于培养转染后的BMSCs。通过CCK-8检测BMSCs增殖活力；运用碱性磷酸酶、茜素红及油红O染色检测各组细胞成骨成脂分化情况；通过qRT-PCR、Western Blot法检测成骨、成脂、MAPK/ P38-ERK信号通路相关基因及蛋白的表达。选取24只SPF级C57BL/6雄性小鼠用于制作股骨横断骨折模型，造模后给予不同转染的BMSCs治疗并给予ZJYXD。通过X-ray与Micro-CT检测对照组、对照组+ZJYXD组、ANRIL过表达组、ANRIL过表达+ZJYXD组小鼠骨愈合情况。结果 与0 %含药血清相比，6 % ZJYXD含药血清对BMSCs细胞增殖能力最强（P<0.01）。ZJYXD干预后成骨染色加深，成骨相关基因OPN、RUNX2 mRNA及蛋白表达显著升高（P<0.01）；成脂染色减弱，相关基因PPARγ、C/EBPα mRNA及蛋白表达显著降低（P<0.01）。同时ZJYXD促进了P-ERK、P38蛋白表达（P<0.01）。X-ray及Micro-CT显示ZJYXD促进了小鼠骨折后愈合。当ANRIL过表达后，ZJYXD调节成骨成脂细胞分化作用消失，无法有效的促进骨折愈合。结论 壮筋养血汤具有促进成骨分化、抑制成脂分化的作用，而这种作用通过LncRNA ANRIL调控MAPK/ P38-ERK信号通路实现。 |
| 英文摘要: |
| Objective To explore the mechanism of Zhuangjin Yangxue Decoction （ZJYXD）in regulating osteogenic and adipogenic differentiation through MAPK/P38-ERK pathway mediated by LncRNA ANRIL. Methods Bone mesenchymal stem cells (BMSCs) were extracted and transfected with ANRIL lentivirus, then ZJYXD containing serum was prepared and used to culture the transfected BMSCs. The proliferation of BMSCs was detected by CCK-8, and the osteogenic differentiation of BMSCs was detected by alkaline phosphatase, alizarin red and oil red O staining; qRT-PCR and Western blot were used to detect the expression of genes and proteins related to osteogenesis, adipogenesis and MAPK/P38-ERK signaling pathway. 24 SPF C57BL/6 male mice were used to establish the model of transverse femoral fracture, after modeling, BMSCs were treated with different transfection and ZJYXD. X-ray and micro-CT were used to detect the bone healing in different groups. Results Compared with 0% ZJYXD-containing serum, 6% ZJYXD-containing serum had the strongest proliferative effect on BMSCs (P<0.01). After ZJYXD intervention, the mRNA and protein expressions of osteogenesis related genes OPN and RUNX2 were significantly increased (P<0.01), while the mRNA and protein expressions of adipogenesis related genes PPARγ and C/EBPα were significantly decreased (P<0.01). At the same time, ZJYXD promoted the expression of P-ERK and P38 protein (P<0.01). X-ray and micro-CT showed that ZJYXD promoted fracture healing in mice. After overexpression of ANRIL, the effect of ZJYXD on osteoblast differentiation disappeared, and it could not effectively promote fracture healing. Conclusion ZJYXD can promote osteogenic differentiation and inhibit adipogenic differentiation, which is accomplished by regulating MAPK/P38-ERK signal pathway by LncRNA ANRIL. |
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