夏枯草颗粒对大鼠增生乳腺组织中 Bcl-2、Bax、PCNA蛋白表达的影响

目的 观察夏枯草颗粒对乳腺增生大鼠乳腺组织中B细胞淋巴瘤-2(B cell lymphoma-2, Bcl-2)、Bcl-2相关X蛋白(Bcl-2 Associated X Protein, Bax)和增殖细胞核抗原(Proliferating Cell Nuclear Antigen, PCNA)蛋白表达的影响，探究夏枯草颗粒发挥作用机制。 方法 雌性育龄期SD大鼠30只，随机数字表法分为正常对照组(n=5)和乳腺增生模型组(n=25)。模型组序贯肌注苯甲酸雌二醇及黄体酮建立乳腺增生模型。建模后将模型组用随机数字表法分为：模型对照组、三苯氧胺组、夏枯草低、中、高剂量组，共5组，每组5只。各组分别予生理盐水、三苯氧胺1.8mg·Kg-1、夏枯草颗粒0.5g·Kg-1、夏枯草颗粒1.0g·Kg-1、夏枯草颗粒3.0g·Kg-1灌胃，共4周。给药结束后，留取组织标本，观察组织学特征，使用免疫组化二步法检测组织中Bcl-2、Bax、PCNA的表达，并用免疫组织化学评分评价Bcl-2、Bax、PCNA表达强弱。结果 平均腺泡导管数：三苯氧胺组、夏枯草低、中、高剂量组与模型对照组相比，平均腺泡导管数明显减少(6.93±2.74)、(10.80±6.42)、(10.47±5.46)、(8.93±5.36)比(15.93±7.68)，均P<0.05]。平均扩张导管数：三苯氧胺组、夏枯草低、中、高剂量组与模型对照组相比，扩张导管数明显减少(2.87±1.19)、(3.13±1.25)、(3.40±1.35)、(2.00±2.39)比(8.73±3.08),均P<0.05]。PCNA、Bcl-2、Bax的免疫组织化学评分：与正常对照组相比，模型对照组大鼠乳腺组织PCNA表达增加，免疫组织化学评分(64.57±27.93) 分比 (92.61±27.36) 分, P<0.05]，Bax表达下降(87.63±25.80) 分 比 (67.08±32.13) 分 , P<0.05]。与模型对照组比较，夏枯草各剂量组PCNA表达明显减少(92.61±27.36)分比 (47.07±31.51) 分、(58.16±37.86) 分、(30.60±32.36) 分, 均P<0.05]。此外，高剂量组Bcl-2表达下降明显(101.91±16.82) 分比(71.19±22.90 ) 分, P<0.05]。结论 夏枯草颗粒具有改善大鼠乳腺增生的作用，其机制可能与下调PNCA表达、调节Bcl-2及Bax表达有关。

Effect of Prunella vulgaris on the expression of Bcl-2, Bax and PCNA protein in hyperplastic mammary gland tissues of rats

Objective To observe the effect of Prunella vulgaris granule on the expression of Bax, Bcl-2 and PCNA in hyperplasia mammary gland tissue of rats. Methods 30 female rats of childbearing age were randomly divided into normal control group (n =5) and mammary gland hyperplasia model group(n=25). The models were injected with estradiol benzoate and progestin. After successful modeling, the mammary gland hyperplasia models were randomly divided into five groups: model control group, tamoxifen group, low, middle and high dose Prunella vulgaris groups, each with 5 animals. Each group was fed with saline, tamoxifen 1.8mg·Kg-1, Prunella vulgaris granule 0.5g·Kg-1, Prunella vulgaris granule 1.0g·Kg-1 and Prunella vulgaris granule 3.0g·Kg-1 respectively for 4 weeks. After administration, the pathological specimens of rat mammary glands were obtained, the pathological features were observed, and the expressions of Bax, Bcl-2 and PCNA in the pathological tissues were detected by Enhance Labelled Polymer System. In addition, Histochemistry score (HS) was used to evaluate the expression of Bax, Bcl-2 and PCNA. Results Average catheters in acinar gland: compared with the model control group, the average decreased significantly in tamoxifen group, low, middle and high dose of Prunella vulgaris granule groups, i.e., (6.93±2.74), (10.80±6.42), (10.47±5.46), (8.93±5.36) vs. (15.93±7.68), all P<0.05]. Average expandable catheters: the number of expandable catheters decreased significantly in tamoxifen group, low, middle and high dose of Prunella vulgaris granule groups, when compared with the model control group, i.e. (2.87±1.19), (3.13±1.25), (3.40±1.35), (2.00±2.39) vs. (8.73±3.08), all P<0.05]. Histochemistry score of PCNA, Bcl-2 and Bax: compared with the normal control group, the expression of PCNA in the mammary tissue of the model control group increased, i.e., HS (64.57±27.93) points vs. (92.61±27.36) points, P<0.05]. The expression of Bax decreased, i.e., (87.63±25.80) points vs. (67.08±32.13) points, P<0.05]. Compared with the model control group, the expression of PCNA in each dose group of Prunella vulgaris granule decreased significantly, i.e., HS (92.61±27.36) vs. (47.07±31.51) points, (58.16±37.86) points, (30.60±32.36) points, all P<0.05]. In addition, the expression of Bcl-2 in the high dose group decreased, i.e., HS (101.91±16.82) points vs (71.19±22.90) points, P<0.05]. Conclusion Prunella vulgaris granule has a good effect on mammary gland hyperplasia in rats, and the mechanism is mainly related to down-regulating the expression of PNCA and regulating the expression of Bcl-2 and Bax.