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目的:观察地榆鞣质对骨髓抑制小鼠的DNA修复蛋白O6-甲基鸟嘌呤DNA甲基转移酶(O6-methylguanine-DNA methyltransferase,MGMT)基因及蛋白表达的影响。方法:40只昆明种小鼠按体质量随机分为正常对照组、模型组、阳性对照组和地榆鞣质组,每组10只。通过腹腔注射环磷酰胺(cyclophosphamide,CTX)诱导建立小鼠骨髓抑制模型。阳性组在CTX诱导3 d后按30μg/kg剂量经腹腔注射集落刺激因子,地榆鞣质组在CTX诱导前3 d开始按20 mg/kg剂量灌胃给予地榆鞣质水溶液,正常对照组和模型组均灌胃等体积纯净水。用药治疗结束后,收集血液和骨髓,FCM法测定骨髓细胞DNA含量,并采用反转录PCR(reverse transcriptionPCR,RT-PCR)和蛋白质印迹法测定骨髓细胞中MGMT基因及蛋白表达的变化。结果:与正常对照组比较,模型组小鼠骨髓DNA含量显著减少(P<0.01),骨髓细胞中MGMT基因及其蛋白表达也均显著减少(P<0.01)。与模型组比较,地榆鞣质可明显提高小鼠骨髓DNA含量(P<0.01),并明显促进骨髓细胞中MGMT基因及蛋白的表达(P<0.01)。结论:地榆鞣质可促进骨髓抑制小鼠骨髓细胞中MGMT基因及蛋白的表达。
Objective: To observe the effect of elm tannin on gene and protein expression of DNA repair protein O6-methylguanine-DNA methyltransferase (MGMT) in myelosuppression mice. Methods: Forty Kunming mice were randomly divided into normal control group, model group, positive control group and elm tannin group. The mouse model of myelosuppression was induced by intraperitoneal injection of cyclophosphamide (CTX). In the positive group, the colony stimulating factor was injected intraperitoneally at a dose of 30μg / kg after 3 days of induction of CTX. At 3 days before the induction of CTX, the trimethine tannin group was given gavage at a dosage of 20 mg / kg. The normal control group And the model group were fed with equal volume of pure water. After treatment, blood and bone marrow were collected. The DNA content of bone marrow cells was determined by FCM. The expression of MGMT gene and protein in bone marrow cells was determined by reverse transcription PCR (RT-PCR) and Western blotting. Results: Compared with the normal control group, the content of bone marrow DNA of model mice decreased significantly (P <0.01), and the expression of MGMT gene and protein in bone marrow cells decreased significantly (P <0.01). Compared with the model group, Burkina Faso tannin significantly increased the content of bone marrow DNA (P <0.01), and significantly promoted the expression of MGMT gene and protein in bone marrow cells (P <0.01). Conclusion: The tannin can promote the expression of MGMT gene and protein in bone marrow cells of myelosuppressed mice.