Microsoft Word _编辑二校

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1 7 8 Vol. 7 No Journal of Food Safety and Quality Aug., 2016 马玉 1, 吕建云 1, 孙丰霞 1, 耿越 1*, 杨长军 2, 王桐 (1., ; 2. ( ), ) 2 摘要 : 目的 (benign prostatic hyperplasia, BPH) 方法 6, BPH 20 mg/kg d 40 mg/kg d 80 mg/kg d (0.8 mg/kg d) 2, 3 4, IL-1 IL-6 TNF- DHT NO NOS PACP ;, 结果 3 BPH DHT PACP NOS, NO 结论 BPH, BPH, 关键词 : ; ; ; ; Preventive effect of Pinus massoniana pollen extract on benign prostate hyperplasia in mice MA Yu 1, LV Jian-Yun 1, SUN Feng-Xia 1, GENG Yue 1*, YANG Chang-Jun 2, WANG Tong 2 (1. Provincial Key Laboratory of Animal Resistant Biology, College of Life Science, Shandong Normal University, Jinan , China; 2. Yantai New Era Health Industry (Group) Co., Ltd., Yantai , China) ABSTRACT: Objective To explore the influences of masson pine pollen extract on benign prostate hyperplasia (BPH) in mice. Methods Kunming male mice were randomly divided into 6 groups, which were normal control group, BPH model group, 20 mg/kg (low), 40 mg/kg (middle), 80 mg/kg(high) masson pine pollen extract groups and Finasteride group (0.8 mg/kg d). Mice of 3 pollen groups were intragastrically administrated with pollen extract for 2 weeks in advance. Using testosterone propionate to build the BPH mice model meanwhile pollen extract and finasteride were intragastrically administrated respectively for 4 weeks continuously. At the end of experiment, the prostate gland and spleen were dissected for determination of prostate index (PI) and spleen index(si), and IL-6, IL-1β, TNF-α, PACP, DHT, NO, NOS and PACP were measured by test kits. Results Three doses of masson pine pollen extract all decreased PI, cytokines, DHT concentration, the activities of PACP and NOS, and reduced the generation of NO. Conclusion Masson pine pollen extract can significantly reduce the pathological symptoms and biochemical parameters of BPH so as to protect BPH in some extent, and the mechanism may be related to the inhibition of inflammation and antioxidant activity. KEY WORDS: masson pine pollen; extract; benign prostatic hyperplasia; inflammatory response; oxidative stress * 通讯作者 :,, gengy@sdnu.edu.cn *Corresponding author: GENG Yue, Professor, Department of Food Science and Technology, Shandong Normal University, Jinan , China. gengy@sdnu.edu.cn

3260 7 1 引言, (benign prostatic hyperplasia, BPH),,, BPH, 5α- [1],, α1- [2], [3] BPH,, [4], BPH 20 80, [5] [6],, [7],,,,,,,, 2 材料与方法 2.

2 引言, (benign prostatic hyperplasia, BPH),,, BPH, 5α- [1],, α1- [2], [3] BPH,, [4], BPH 20 80, [5] [6],, [7],,,,,,,, 2 材料与方法 2.1 材料与仪器 (masson pine pollen extract, MPPE), ( ) : (Pinus massoniana),,, ph,,, 4 (20±2) g, SPF ; ( ), (L/N: );, (L/N:121203); Elisa (IL-1 IL-6 TNF- PACP DHT) R&D (L/N:201312) NO/NOS, (L/N: ) Centrifuge 5804R ( Eppendorf ) 2, 3 MPPE,, 0.2 ml/, 3,, 5 mg/kg d,, MPPE 2.3 样品收集与处理 6,, EP, EP 1 h, 4, 2, -20, ELISA IL-1 IL-6 TNF- (dihydrotestosterone, DHT) NO NOS (prostatic acid phosphatase PACP),, (prostate index, PI) (spleen index, SI), HE (10 mg/g) 10 (mg)/ (g); (mg/g) (mg)/ (g) 2.4 数据分析 X S, SPSS, 3 结果与分析 3.1 小鼠前列腺指数和脾脏指数的变化 4, PI (P 0.01)( 1), 4 PI, PI(P 0.05 P 0.01),, SI (P 0.01), 分组设计与方法, 6 : (Normal control) (BPH model) 20 mg/kg d (Low MPPE) 40 mg/kg d(middle MPPE) 80 mg/kg d(high MPPE) (Finasteride, 0.8 mg/kg d), 6 1 PI SI (n=6) Fig. 1 Changes of PI and SI in mice of each group (n=6)

mg/kg(见图 3 一定差异, 总体呈现剂量依赖效应关系 E) MPPE 组的前列腺增生得到明显改善, Finasteride 组的腺泡结构基本正常(见图 3F) 3.4 小鼠血清中 DHT 的含量变化实验结束时, 模型组小鼠血清的 DHT 含量升高(P 0.01), 见图 4 非那雄胺(P 0.01)以及中(P 0.05) 高剂量(P 0.

3 第8期马玉, 等: 马尾松花粉提取物对小鼠前列腺肥大的预防作用血清中细胞因子含量(IL-1 IL-6 TNF- )测定到腺泡腔, 形成增生结节上皮细胞呈柱状, 细胞排列不规与空白组相比, 模型组小鼠血清中的 3 种前炎性细胞则(见图 3B) 20 mg/kg MPPE 组的前列腺增生情况较重, 上因子含量均明显增加(图 2), 3 种剂量的 MPPE 降低程度有皮增生明显(见图 3C), 40 mg/kg(见图 3D) 80 mg/kg(见图 3 一定差异, 总体呈现剂量依赖效应关系 E) MPPE 组的前列腺增生得到明显改善, Finasteride 组的腺泡结构基本正常(见图 3F) 3.4 小鼠血清中 DHT 的含量变化实验结束时, 模型组小鼠血清的 DHT 含量升高(P 0.01), 见图 4 非那雄胺(P 0.01)以及中(P 0.05) 高剂量(P 0.01)的 MPPE 均降低了血清中 DHT 的含量; 但与模型组相比, 低剂量 MPPE 组 DHT 含量变化不明显, MPPE 的作用有一定的剂量依赖性 3.5 小鼠前列腺组织中 PACP 活性的变化如图 5 所示, 与正常小鼠相比, BPH 组(P 0.01)和低剂量 MPPE 组(P 0.05)小鼠前列腺组织中的 PACP 活性都显著增加与低剂量 MPPE 不同的是, 中(P 0.05) 高剂图 2 小鼠血清中细胞因子的含量(n=6) Fig. 2 Content of cytokines in mice serum (n=6) 注: # P<0.05, ## P<0.01 vs 空白组; * P<0.05, ** P<0.01 vs 模型组量(P 0.01)MPPE 发挥了明显的保护作用, 非那雄胺的保护是最有效的 3.6 血清中 NO, NOS (inos, TNOS)的含量如图 6 所示, 与空白组相比, 模型组小鼠血清中的小鼠前列腺组织形态学检查 NO 含量显著增加(P 0.01), 2 种 NOS 的活性同样明显增小鼠前列腺组织形态学结果见图 3 正常对照组腺泡强经过 4 周的干预, 非那雄胺起到了显著的效果(P 上皮细胞呈单层, 形状规则核圆, 排列整齐(见图 3A) 0.01), 3 个松花粉组 NO 含量及 NOS 活性也明显降低(P BPH 组小鼠前列腺腺泡形状不规则, 上皮细胞绒毛状突起 0.01), 并且 3 个剂量之间无显著性差异 3.3 图 3 各组小鼠前列腺组织形态学检查(HE, 200 ) Fig. 3 Morphology results of prostate tissue in different groups of mice A: Normal control; B: BPH model; C: Low MPPE; D: Middle MPPE; E: High MPPE; F: Finasteride

tissue of each group (n=6) 6 NO NOS (n=6) Fig.

Finasteride, BPH [8-11], [12] EPA DHA PGE2

4 DHT (n=6) Fig. 4 Changes of DHT concentration in mice serum of each group(n=6) 5 PACP (n=6) Fig.5 Changes of PACP activities in mice prostate tissue of each group (n=6) 6 NO NOS (n=6) Fig. 6 Concentration changes of NO and NOS in mice serum of each group (n=6) A.NO; B. inos, TNOS 4 讨论,, BPH,, MPPE, 80 mg/kg MPPE, Finasteride, BPH [8-11], [12] EPA DHA PGE2 IL-6, Mirnikjoo [13], IL-1β, IL-6 TNF-α,, 2,, MPPE BPH,,, [14], DHT (T) 5α- DHT, DHT T 5 ; Ali [15], T DHT,

5 8, : 3263 DHT, DHT, T BPH PACP, PACP [16], PACP, MPPE PACP,, MPPE,,,,, [17] NO [18] inos, NO,,, [19,20] NO, [21] [22], NO, NO, NO,,,, BPH,,,, BPH 参考文献 [1] Carson CR, Rittmaster R. The role of dihydrotestosterone in benign prostatic hyperplasia [J]. Urology, 2003, 61(4 Suppl 1): 2 7. [2] Ripple MO, Henry WF, Rago RP, et al. Prooxidant-antioxidant shift induced by androgen treatment of human prostate carcinoma cells [J]. J Natl Cancer Inst, 1997, 89(1): [3] Veeresh BS, Veeresh B, Patil AA, et al. Lauric acid and myristic acid prevent testosterone induced prostatic hyperplasia in rats [J]. Eur J Pharmacol, 2010, 626(2 3): [4],,. [J]., 2003, (02): 3 5. ZhangL, Feng NP, Wu CL. Comparison of Chinese and western medicine in the treatment of benign prostatic hyperplasia [J]. Chin J Inf Tradit Med, 2003, (02): 3 5. [5]. [J]., 1989, (02): Qian BC. New progress in research on nutrition and pharmacology of pollen [J]. Chin J Integr Tradit Chin West Med, 1989, (02): [6],,. [J]., 2005, (05): Cong T, Zhao L, Bao SF. The effect of pine pollen on the treatment of benign prostatic hyperplasia in rats [J]. Acad J PLA Postgrad Med Sch, 2005, (05): [7],,. [J]., 1998, (11): Du XZ, Zhang JY, Wang GL, Current status and progress of drugs for the treatment of benign prostatic hyperplasia [J]. China Pharm, 1998, (11): [8] Penna G, Mondaini N, Amuchastegui S, et al. Seminal plasma cytokines and chemokines in prostate inflammation: interleukin 8 as a predictive biomarker in chronic prostatitis/chronic pelvic pain syndrome and benign prostatic hyperplasia [J]. Eur Urol, 2007, 51(2): , 533. [9] Wang W, Bergh A, Damber JE. Chronic inflammation in benign prostate hyperplasia is associated with focal upregulation of cyclooxygenase-2, Bcl-2, and cell proliferation in the glandular epithelium [J]. Prostate, 2004, 61(1): [10],,. IL-10 TNF-α [J]. ( ), 2012, (01): Sun GH, NIng FX, Xia YJ. Expression and significance of IL-10 and TNF-α in benign prostatic hyperplasia and prostate cancer [J]. J Urol Clin (ElectrVer), 2012, (01): [11],. CD8 CD20 IL-6 IL-23 [J]., 2011, (08): Zhang W, Han RF. Expression and Significance of CD8, CD20, IL-6 and IL-23 in tissue of benign prostatic hyperplasia [J]. Tianjin Med J, 2011, (08): [12],,. EPA DHA PGE_2 IL-6 IL-6 mrna [J]., 2006, (04): Xia YP, Su YX, Feng X. Effects of EPA, DHA on secretion of PGE2, IL-6 and Expression of IL-6m RNA in human monocytes [J]. Acta Nutr Sin, 2006, (04): [13] Mirnikjoo B, Brown SE, Kim HF, et al. Protein kinase inhibition by omega-3 fatty acids [J]. J Biol Chem, 2001, 276(14): [14] Hobisch A, Eder I E, Putz T, et al. Interleukin-6 regulates prostate-specific protein expression in prostate carcinoma cells by activation of the androgen receptor [J]. Cancer Res, 1998, 58(20): [15] Ali MI, Kondreddi HD, Veeresh B. Protective effect of 2-hydroxy-4-methoxy benzoic acid on testosterone induced benign prostatic hyperplasia in Wister rats [J]. Eur J Pharm, 2013, 698(1-3): [16],. T E_2 [J]., 2006, (03): Liao ZY, Jiang JL. The effect of Qianlie granute on serum PAP and T & E2 in model of prostatic hyperplasis rats [J]. Prac J Med Pharm, 2006, (03): [17] Cheeseman KH. Mechanisms and effects of lipid peroxidation [J]. Mol Aspects Med, 1993, 14(3): [18],. [J]., 2010, (09): Luo JH, Yang YB. Protective effects of resveratrol on alcohol-induced hepatic injury through nitric oxide pathway in rats [J]. Chin J Mod Appl Pharm, 2010, (09): [19] Rikitake Y, Hirata K, Kawashima S, et al. Involvement of endothelial

6 nitric oxide in sphingosine-1-phosphate-induced angiogenesis [J]. Arterioscler Thromb Vasc Biol, 2002, 22(1): [20],,,. [J]., 2009, (11): Tian YT, Zeng ZY, Chen WW, et al. The influence of polyic on blood capillary in mouse prostate carcinoma [J]. Chin Pharm Bull, 2009, (11): [21],,,. NF-κB-iNOS-NO PC12 [J]., 2009, (07): Kuang R, Sun YG, Zheng XX, et al. Lipoic acid protected PC12 cells from oxidative stress-induced damage by inhibiting the Nf-κBiNOS-NO signal pathway [J]. Chin J Mod Appl Pharm, 2009, (07): [22]. [J]., 2013, (12): Lin XT. Objective to investigate the value of oxidative stress in the pathogenesis of benign prostatic hyperplasia and prostate cancer [J]. Guide China Med, 2013, (12): 作者简介 ( 责任编辑 : 姚菲 ) 马玉, 硕士研究生, 主要研究方向为食品科学 m88y0701@sina.com 耿越, 教授, 主要研究方向为食品科学 gengy@sdnu.edu.cn

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Microsoft Word _编辑 二校

Microsoft Word _编辑二校