莪术油对大鼠乳腺癌癌前病变组织血管生成及相关调控因子表达的研究
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摘要
目的：本研究以活血化瘀中药莪术有效成份莪术油为干预药物，从大鼠机体生存状态、组织病理形态学方面评价莪术油对大鼠乳腺癌癌前病变动物模型的逆转效应，从血管生成（微血管密度MVD、VEGF）角度，应用组织病理学、免疫组化、原位杂交等检测方法，从全身、组织、细胞及分子等多个层面，探讨莪术油对大鼠乳腺癌癌前病变组织血管生成及相关调控因子VEGFmRNA表达的相关作用机制。方法：275只SD大鼠随机分为空白对照组、造模对照组、他莫昔芬组、康莱特组及莪术油小、中、大剂量组。采用二甲基苯蒽（DMBA）诱导乳腺癌癌前病变造模。实验开始后定期观察大鼠生物学行为的变化，干预治疗4周，第8～14周分4批处死动物，观察各组大鼠乳房外形、进行病理学检测、采用免疫组化法测定标本中CD34标记的微血管密度（MVD）表达情况、采用原位杂交法测定标本中VEGFmRNA表达情况。结果：1、造模对照组与空白对照组比较，大鼠死亡率、癌前病变发生率增高（P＜0.05）。莪术油大、中剂量组癌前病变发生率降低，与病模组及康莱特组比较差异显著（P＜0.05），与他莫西芬组比较无显著差异（P>0.05）。2、各实验组不同类型乳腺组织中MVD、VEGFmRNA表达阳性率呈递增趋势。3、非典型增生乳腺组中，莪术油中、大剂量组、他莫昔芬组MVD表达阳性率均明显低于造模对照组(P<0.05)，各干预组的VEGFmRNA阳性细胞阳性率均明显低于造模对照组（P<0.05），莪术油组低于康莱特和他莫昔芬组（P<0.05）。对VEGFmRNA表达阳性的非典型增生标本，莪术油大剂量组VEGFmRNA阳性细胞阳性率明显低于小剂量组（P<0.05）。结论：DMBA诱导大鼠乳腺癌癌前病变模型，能够复制出乳腺增生到癌变过程中各个阶段的病理改变。在单纯乳腺增生→乳腺癌癌前病变→乳腺癌的演变进程中，癌前病变阶段MVD、VEGFmRna表达呈现显著性增高，说明血管生成及相关调控因子能反映乳腺癌癌前病变的恶性倾向。莪术油能够显著抑制乳腺癌癌前病变大鼠乳腺血管生成及相关调控因子MVD蛋白的表达，有效的降低DMBA诱导大鼠乳腺癌前病变组织中VEGFmRNA表达强度，可能是其抑制血管生成及阻断乳腺癌发生的有效机制。
Objective: the study evaluate reversal effect of Zedoray Turmeric oil on rat breastprecancerous lesions animal models, from rat survival of the state of the body, pathologicalmorphological aspects. From angiogenesis (microvessel density MVD, VEGF), withdetection method Histopathology, immunohistochemistry, in situ hybridization, exploreeffect and mechanism of the the Zedoray Turmeric oil to reverse breast precancerouslesions and Expression of Related Regulatory Factors VEGF mRNA in animal body of rats,in systemic, tissue, cell and molecular level. Methods：275SD rats were randomly dividedinto7groups: the blankcontrol group(BC),the precancerous lesion model controlgroup(PL),the kanglaite control group(KLT),the tamoxifen control group(TAM),the lowdose group of ZTO(ZTO-L),the middle dose group of ZTO(ZTO-M) and the high dosegroup of ZTO(ZTO-H). Animal model of precancerous lesion of rats mammary wasinduced bye DMBA and was treated by ZTO,KLT and TAM for4weeks. All the animalswere killed from8th to14th week once every two weeks. Observe breast shape,pathologydetection in each group, Determinate the microvascular markered with CD34density usingmmunohistochemical methods in the specimens,examination of histopathology wasperformed and measured the expression of VEGF mRNA by in situ hybridization. Results:1. The disease model group compared with the blank control group,rat mortality andincidence of precancerous lesions increased (P <0.05). the incidence of precancerous lesionof the high-dose group and the middle dose of Zedoray Turmeric oil reduced comparedwith disease module group and KLT group showed significant difference (P <0.05),compared with the tamoxifen control group show no significant difference (P>0.05).2.The positive rate of expression of VEGF mRNA and MVD trend to increase in every groups except BC corresponding to normal，usual ductal hyperplasia(UDH),atypical ductalhyperplasia(ADH) and invasive carcinoma.3. In ADH, The positive-cell rate of MVD ofZTO-M, ZTO-H and TAM was much lower than PL(P<0.05),the positive-cell rate ofVEGF mRNA of ZTO, KLT and TAM was much lower than PL(P<0.05),and thepositive-cell rate of VEGF mRNA of ZTO was much lower than KLT and TAM(P<0.05).In the case of ADH which the expression of VEGF mRNA was positive，the positive-cellrate of VEGF mRNA of ZTO-H was was much lower than ZTO-L(P<0.05). Conclusion:The breast precancerous lesion model induced by DMBA is able to replicate thepathological changes of the various stages in the process of mammary gland hyperplasia tocarcinoma. In the evolution process of simple mammary hyperplasia→breastprecancerous lesions→breast cancer, the MVD, VEGFmRna expression presentedsignificantly higher in the stage of precancerous lesions, this show that angiogenesis andrelated regulatory factor can reflect the vicious tendency of precancerous lesions of breastcancer. edoray Turmeric Oil decrease the expression of VEGF mRNA in precancerouslesion of rats mammary induced by DMBA, which could be the mechanism to inhibitsangiogenesis and prevent the breast cancer.
Objective: the study evaluate reversal effect of Zedoray Turmeric oil on rat breastprecancerous lesions animal models, from rat survival of the state of the body, pathologicalmorphological aspects. From angiogenesis (microvessel density MVD, VEGF), withdetection method Histopathology, immunohistochemistry, in situ hybridization, exploreeffect and mechanism of the the Zedoray Turmeric oil to reverse breast precancerouslesions and Expression of Related Regulatory Factors VEGF mRNA in animal body of rats,in systemic, tissue, cell and molecular level. Methods：275SD rats were randomly dividedinto7groups: the blankcontrol group(BC),the precancerous lesion model controlgroup(PL),the kanglaite control group(KLT),the tamoxifen control group(TAM),the lowdose group of ZTO(ZTO-L),the middle dose group of ZTO(ZTO-M) and the high dosegroup of ZTO(ZTO-H). Animal model of precancerous lesion of rats mammary wasinduced bye DMBA and was treated by ZTO,KLT and TAM for4weeks. All the animalswere killed from8th to14th week once every two weeks. Observe breast shape,pathologydetection in each group, Determinate the microvascular markered with CD34density usingmmunohistochemical methods in the specimens,examination of histopathology wasperformed and measured the expression of VEGF mRNA by in situ hybridization. Results:1. The disease model group compared with the blank control group,rat mortality andincidence of precancerous lesions increased (P <0.05). the incidence of precancerous lesionof the high-dose group and the middle dose of Zedoray Turmeric oil reduced comparedwith disease module group and KLT group showed significant difference (P <0.05),compared with the tamoxifen control group show no significant difference (P>0.05).2.The positive rate of expression of VEGF mRNA and MVD trend to increase in every groups except BC corresponding to normal，usual ductal hyperplasia(UDH),atypical ductalhyperplasia(ADH) and invasive carcinoma.3. In ADH, The positive-cell rate of MVD ofZTO-M, ZTO-H and TAM was much lower than PL(P<0.05),the positive-cell rate ofVEGF mRNA of ZTO, KLT and TAM was much lower than PL(P<0.05),and thepositive-cell rate of VEGF mRNA of ZTO was much lower than KLT and TAM(P<0.05).In the case of ADH which the expression of VEGF mRNA was positive，the positive-cellrate of VEGF mRNA of ZTO-H was was much lower than ZTO-L(P<0.05). Conclusion:The breast precancerous lesion model induced by DMBA is able to replicate thepathological changes of the various stages in the process of mammary gland hyperplasia tocarcinoma. In the evolution process of simple mammary hyperplasia→breastprecancerous lesions→breast cancer, the MVD, VEGFmRna expression presentedsignificantly higher in the stage of precancerous lesions, this show that angiogenesis andrelated regulatory factor can reflect the vicious tendency of precancerous lesions of breastcancer. edoray Turmeric Oil decrease the expression of VEGF mRNA in precancerouslesion of rats mammary induced by DMBA, which could be the mechanism to inhibitsangiogenesis and prevent the breast cancer.
引文
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