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目的:探讨龟龙坚骨汤对TNF-α诱导大鼠成骨细胞凋亡中Fas和Bcl-2表达的影响。方法:将原代培养的SD大鼠颅骨成骨细胞分为中药组和对照组,中药组给予龟龙坚骨汤含药血清培养,对照组给予常规血清培养,培养7d后,用含量为30ng/mL TNF-α刺激成骨细胞,采用流式细胞术检测细胞凋亡,应用免疫组化结合计算机图像分析技术检测细胞Fas和Bcl-2蛋白表达。结果:TNF-α刺激成骨细胞后,中药组中Bcl-2蛋白表达升高,Fas蛋白表达不明显;而对照组的Fas蛋白表达升高,Bcl-2蛋白表达不明显。在细胞凋亡率方面,中药组在早期及晚期细胞凋亡率均低于对照组,两组差异均有统计学意义(P<0.05)。结论:龟龙坚骨汤能上调Bcl-2蛋白表达,对Fas蛋白表达不明显,在一定程度上可以抑制成骨细胞的凋亡,但能否促进成骨细胞的增殖,有待下一步的研究。
Objective: To investigate the effect of Guilonglongjian decoction on the expression of Fas and Bcl-2 in rat osteoblasts induced by TNF-α. Methods: The primary cultured osteoblasts of SD rats were divided into two groups: Chinese medicine group and control group. The traditional Chinese medicine group was given serum containing Guilongjianjian decoction, while the control group was given conventional serum culture. After cultured for 7 days, / mL TNF-α stimulated osteoblasts, apoptosis was detected by flow cytometry, immunohistochemistry and computer image analysis of Fas and Bcl-2 protein expression. Results: After osteoblasts were stimulated with TNF-α, the expression of Bcl-2 protein and Fas protein were not significantly increased in the Chinese herbal medicine group. However, the expression of Fas protein in the control group increased but the expression of Bcl-2 protein was not significant. In the aspect of apoptosis rate, the apoptotic rate of the TCM group in early and late stages was lower than that in the control group, with significant difference between the two groups (P <0.05). Conclusion: Guilongjian decoction can up-regulate the expression of Bcl-2 protein but not Fas protein, which can inhibit the apoptosis of osteoblasts to a certain extent, but whether it can promote the proliferation of osteoblasts remains to be further studied .