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目的:研究益肾解毒方对小鼠溶骨性骨转移瘤细胞核糖核酸酶L(ribonuclease L,RNase L)蛋白及凋亡终末蛋白半胱氨酸蛋白酶-3(Caspase-3)蛋白表达的调控作用。方法:将已经获得国家专利的肺腺癌溶骨性骨转移细胞株,给予裸鼠左心室注射,制备溶骨性骨转移模型,随机分为5组,益肾解毒方高、中、低剂量(24,12,3 g·kg~(-1))组,帕米磷酸二钠组及模型组,以Micro SPECT/CT显像及病理组织学方法观察骨转移发生情况,定时定量给药并于5周后取材,通过免疫组织化学、蛋白免疫印迹法(Western blot)以及反转录聚合酶链式反应(RT-PCR)等方法研究RNase L,Caspase-3及益肾解毒方的干预作用。结果:与模型组比较,帕米磷酸二钠组、益肾解毒方高、中、低剂量组骨转移瘤细胞RNase L蛋白,mRNA表达降低(P<0.01),益肾解毒方中、低剂量组及帕米磷酸二钠组骨转移灶中Caspase-3蛋白,mRNA表达升高(P<0.01)。讨论:肺腺癌骨转移灶中RNase L高表达,而细胞凋亡过程中最主要的终末剪切酶Caspase-3表达降低,其内在机制可能与肺腺癌骨转移细胞的凋亡逃避机制相关,益肾解毒方能够降低RNase L表达,增强Caspase-3表达,一定程度上抑制了肿瘤细胞的凋亡逃避,这可能是益肾解毒方抗肿瘤的内在机制。
Objective: To study the effect of Yishenjiedu decoction on the expression of ribonuclease L (RNase L) protein and the apoptotic caspase-3 protein in mouse osteosarcoma bone metastases Regulation. Methods: The lung adenocarcinoma osteolytic bone metastasis cell line, which has obtained the national patent, was injected into the left ventricle of nude mice to prepare the osteolytic bone metastasis model and randomly divided into 5 groups: Yishenjiedu Fang high, medium and low dose (24, 12, 3 g · kg ~ (-1)) group, Pamidronate disodium group and model group. The occurrence of bone metastasis was observed by Micro SPECT / CT imaging and histopathology. After 5 weeks, the interventions of RNase L, Caspase-3 and Yishen Jiedu were studied by immunohistochemistry, Western blot and reverse transcriptase-polymerase chain reaction (RT-PCR) . Results: Compared with the model group, the expressions of RNase L protein and mRNA of bone metastasis cells in Pamidronate disodium, Yishenjiedu high, medium and low dose groups decreased (P <0.01) Caspase-3 protein and mRNA expression in bone metastases of Pamidronate group and Pamidronate group were significantly increased (P <0.01). DISCUSSION: RNase L is highly expressed in bone metastases of lung adenocarcinoma, while the expression of Caspase-3, the most important terminal cleavage enzyme, is decreased in the process of apoptosis. The intrinsic mechanism may be related to the mechanism of apoptosis escape from bone metastases of lung adenocarcinoma Yishen Jiedu Decoction can reduce RNase L expression and enhance Caspase-3 expression, to a certain extent, inhibit the tumor cells from escaping, which may be Yishenjiedu Fang anti-tumor mechanism.