Jiawei Yanghe Decoction-containing serum attenuates hydrogen peroxide-induced oxidative damage in osteoblasts

doi:10.12307/2026.476

Abstract

Abstract: BACKGROUND: Activation of nuclear factor erythroid 2-related factor 2 may be an effective strategy for antioxidant stress in osteoblasts and prevention of osteoporosis.
OBJECTIVE: To investigate the protective effect and mechanism of Jiawei Yanghe Decoction-containing serum on hydrogen peroxide-induced osteoblast injury.
METHODS: The mouse osteoblast cell line MC3T3-E1 was divided into five groups: control group, model group, and low-, medium-, and high-dose Jiawei Yanghe Decoction-containing serum groups. Cells in the control group were routinely cultured. Cells in the model group were treated with hydrogen peroxide to establish an oxidative stress injury model of osteoblasts. Cells in the low-, medium-, and high-dose Jiawei Yanghe Decoction-containing serum groups were treated with final volume fractions of 5%, 10%, and 15% Jiawei Yanghe Decoction-containing serum for 24 hours based on the model group. Cell status, osteogenic function, oxidative stress level, inflammatory response, and protein expression related to the nuclear factor erythroid 2-related factor 2/heme oxygenase 1 signaling pathway were detected.
RESULTS AND CONCLUSION: Compared with the control group, the model group showed significantly decreased cell viability, alkaline phosphatase activity, osteogenic mineralization capacity, antioxidant enzyme activities, and protein expression levels of nuclear factor erythroid 2-related factor 2 and heme oxygenase 1, and significantly increased apoptosis rate, reactive oxygen species levels, and levels of the inflammatory factors interleukin-6 and interleukin-1β (P < 0.05). Compared with the model group, the Jiawei Yanghe Decoction-containing serum groups showed varying degrees of improvement in the above indicators (P < 0.05). These findings indicate that Jiawei Yanghe Decoction-containing serum can restore the viability and bone formation function of MC3T3-E1 cells and alleviate oxidative stress injury by activating the nuclear factor erythroid 2-related factor 2/heme oxygenase 1 antioxidant signaling pathway.

Key words: Jiawei Yanghe Decoction, MC3T3-E1 cell, osteoblast, nuclear factor erythroid 2-related factor 2, heme oxygenase 1, osteogenic differentiation, oxidative stress, apoptosis

CLC Number:

Wu Fan, Yang Wenlong, Liu Jiangyuan, Xia Hanting, Zeng Zhikui, Yang Fengyun. Jiawei Yanghe Decoction-containing serum attenuates hydrogen peroxide-induced oxidative damage in osteoblasts[J]. Chinese Journal of Tissue Engineering Research, 2026, 30(33): 8583-8590.

Figures/Tables 5

2.1 加味阳和汤含药血清体积分数筛选结果如图1A所示，对照组与体积分数2.5%，5%，10%，15%，20%，25%加味阳和汤含药血清组细胞活力差异均有显著性意义(P < 0.05)。体积分数2.5%-20%加味阳和汤含药血清提高MC3T3-E1细胞细胞活力，其中体积分数15%加味阳和汤含药血清组最佳，而体积分数25%加味阳和汤含药血清具有一定的细胞毒性。如图1B所示，体积分数5%，10%，15%加味阳和汤含药血清均可升高过氧化氢诱导后的MC3T3-E1细胞活力(P < 0.05)。因此选择体积分数5%，10%，15%加味阳和汤含药血清进行后续实验。 2.2 细胞增殖检测结果如图2所示，与对照组比较，模型组MC3T3-E1细胞活力显著降低(P < 0.05)；与模型组比较，加味阳和汤含药血清(体积分数5%，10%，15%)组MC3T3-E1细胞活力显著升高(P < 0.05)。 2.3 细胞凋亡检测结果如图3所示，与对照组(4.68±0.32)%相比，模型组MC3T3-E1细胞凋亡率(64.13±6.98)%明显升高(P < 0.05)；而与模型组相比，加味阳和汤含药血清(体积分数5%，10%，15%)组MC3T3-E1细胞凋亡率则显著降低(P < 0.05)。 2.4 细胞成骨分化与矿化检测结果如图4所示，与对照组比较，模型组碱性磷酸酶染色阳性面积、茜素红染色阳性面积均显著降低(P < 0.05)。与模型组比较，加味阳和汤含药血清(体积分数5%，10%，15%)组碱性磷酸酶染色阳性面积、茜素红染色阳性面积均显著升高(P < 0.05)。 2.5 细胞活性氧水平如图5所示，与对照组(10.53±0.90)%比较，模型组MC3T3-E1细胞活性氧水平(75.50±6.35)%显著升高(P < 0.05)；与模型组比较，加味阳和汤含药血清(体积分数5%，10%，15%)组细胞活性氧水平显著降低(P < 0.05)。 2.6 细胞中抗氧化酶活性与炎症因子水平如图6所示，与对照组比较，模型组过氧化氢酶、超氧化物歧化酶活性均显著降低(P < 0.05)，白细胞介素6、白细胞介素1β水平均显著升高(P < 0.05)。与模型组比较，加味阳和汤含药血清(体积分数5%，10%，15%)组过氧化氢酶、超氧化物歧化酶活性均显著升高(P < 0.05)，白细胞介素6、白细胞介素1β水平均显著降低(P < 0.05)。"

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