麝香配伍乳香促虎杖提取物治疗慢性非细菌性前列腺炎的动物实验研究

Musk and carterii birdw enhance the effect of polygonum extract on chronic non-bacterial prostatitis:An animal experimental study

目的:观察麝香、乳香配伍组合对虎杖提取物治疗慢性非细菌性前列腺炎模型大鼠前列腺组织病理及炎症因子表达的影响。方法:53只健康雄性Wistar大鼠,5只用于大鼠前列腺蛋白提纯液的制备,48只随机分为麝香配伍乳香+虎杖提取物组、虎杖提取物、模型组、正常对照组共4组。除正常对照组外,应用大鼠前列腺蛋白提纯液辅以免疫佐剂制备实验性慢性非细菌性前列腺炎大鼠模型,造模60 d时正常对照组、模型组予生理盐水,虎杖提取物、麝香配伍乳香+虎杖提取物组分别予生药量虎杖1.575 g/(kg.d)、麝香0.021 g/(kg.d)、乳香1.05 g/(kg.d)按组别选用连续灌胃,各组给药14 d时处死,检测指标包括病理学,ELISA法检测前列腺组织匀浆TNF-α、IL-1β、IL-6、IL-8水平,RT-PCR及Western印迹方法检测炎症因子MCP-1(CCL2)、CCR2 mRNA及蛋白表达。结果:麝香配伍乳香+虎杖提取物组前列腺组织结构改善,无明显炎性细胞浸润,虎杖提取物组可见炎性细胞浸润。麝香配伍乳香+虎杖提取物组明显降低前列腺组织炎症因子(TNF-α:11.04±4.07、IL-1β:16.94±4.26、IL-6:110.08±28.42、IL-8:26.28±7.36,pg/ml),优于单纯虎杖提取物组(TNF-α:63.21±21.37、IL-1β:41.32±14.62、IL-6:177.64±42.65、IL-8:96.37±37.61,pg/ml),差异有统计学意义(P<0.05,P<0.01)。麝香配伍乳香+虎杖提取物组大鼠前列腺组织炎症趋化因子MCP-1(CCL2)及其受体CCR2的mRNA(MCP-1:0.32±0.17;CCR2:0.28±0.11)及蛋白(MCP-1:0.28±0.15;CCR2:0.11±0.04)表达水平与模型组(MCP-1 mRNA:1.15±0.39;MCP-1蛋白:0.93±0.34;CCR2 mRNA:0.83±0.26;CCR2蛋白:0.93±0.34)比较显著降低(P<0.01),显著优于虎杖提取物组(MCP-1 mRNA:0.65±0.27;MCP-1蛋白:0.56±0.22;CCR2 mRNA:0.78±0.24;CCR2蛋白:0.25±0.09),差异有统计学意义(P<0.05,P<0.01)。结论:麝香乳香配伍组合的应用具有促进虎杖提取物对慢性前列腺炎的治疗,降低炎症反应程度,从而促进前列腺组织结构修复的作用。

Objective： To observe the effects of musk and carterii birdw on the pathology and the expressions of inflammatory cytokines in chronic non-bacterial prostatitis（CNBP） rats treated with polygonum extract.Methods： Five male Wistar rats were used for the preparation of SC purified prostate protein solution,and another 48 randomly divided into four groups： polygonum extract,polygonum extract ＋ musk and carterii birdw,CNBP model control and normal control.CNBP models were established by injecting SC purified prostate protein solution and Freund＇s complete adjuvant.At 60 days after modeling,the CNBP model control and normal control rats were treated with normal saline,and the other two groups with polygonum extract and polygonum extract ＋ musk and carterii birdw,respectively（polygonum 1.575 g per kg per d,musk 0.021 g per kg per d,and carterii birdw 1.05 g per kg per d）.After 14 days of continuous intragastric medication,all the rats were sacrificed for pathological examination,determination of the levels of TNF-α,IL-1β,IL-6 and IL-8 in the prostate tissue homogenate by ELISA,and detection of the mRNA and protein expressions of inflammatory cytokines MCP-1（CCL2） and CCR2 by RT-PCR and Western blot.Results： The polygonum extract ＋ musk and carterii birdw group showed apparent improvement in the structure of the prostate tissue but no inflammatory infiltration,as was quite obvious in the polygonum extract group.Polygonum extract ＋ musk and carterii birdw significantly decreased the inflammatory cytokines TNF-α（[11.04±4.07] pg/ml）,IL-1β（[16.94±4.26] pg/ml）,IL-6（[110.08±28.42] pg/ml） and IL-8（[26.28±7.36] pg/ml） in the prostate tissue,as compared with polygonum extract alone（[63.21±21.37] pg/ml,[41.32±14.62] pg/ml,[177.64±42.65] pg/ml and [96.37±37.61] pg/ml）（P 0.05,P 0.01）.The former also exhibited significantly lower expressions of MCP-1 mRNA（0.32±0.17）,MCP-1 protein（0.28±0.15）,CCR2 mRNA（0.28±0.11） and CCR2 protein（0.11±0.04） than either the model control group（1.15±0.39,0.93±0.34,0.83±0.26 and 0.93±0.34）（P0.01）,or the polygonum extract group（0.65±0.27,0.56±0.22,0.78±0.24 and 0.25±0.09）（P 0.05,P 0.01）.Conclusion： Musk and carterii birdw can enhance the effect of polygonum extract on chronic prostatitis,reduce inflammatory response and improve tissue repair of the prostate in rats.