白花蛇舌草提取物对CNP大鼠前列腺TLR4/NF-κB通路的影响
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摘要
目的探究白花蛇舌草提取物对慢性非细菌性前列腺炎(CNP)大鼠前列腺toll样受体4(TLR4)/核转录因子(NF-κB)通路的影响。方法抽取SD大鼠80只随机分为空白组、模型组以及白花蛇舌草提取物低、中、高剂量组。通过向前列腺腹侧叶注入完全弗氏佐剂的方法建立CNP模型。造模14 d后开始灌胃给药,白花蛇舌草提取物低、中、高剂量组给药量分别为75、150、300 mg/kg;空白组给予等量生理盐水。给药3周后,观察大鼠前列腺形态和白细胞数目;蛋白质印迹(Western blot)技术检测前列腺组织TLR4、NF-κB水平。结果模型组大鼠体重显著低于空白组,差异有统计学意义(P<0.05),白花蛇舌草提取物各干预组体重与空白组间的差异无统计学意义(P>0.05);白花蛇舌草提取物各干预组前列腺湿重、前列腺湿重/体重比显著高于空白组,差异有统计学意义(P<0.05),且随着给药浓度的升高该比值逐渐降低,差异有统计学意义(P<0.05)。高倍镜下结果显示,各组大鼠前列腺白细胞数目评级差异有统计学意义(P=0.000)。Western blot结果显示,白花蛇舌草提取物干预后前列腺组织TLR4、NF-κB p65蛋白表达水平显著降低(P<0.05),且随着给药浓度的升高,TLR4、NF-κB p65蛋白表达水平逐渐降低(P<0.05)。结论白花蛇舌草提取物能明显减轻CNP大鼠的前列腺损伤程度,其机制可能与TLR4/NF-κB通路受到抑制有关。
Objective To explore the influence of Oldenlandia diffusa extract on prostate toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)pathway of rats with chronic nonbacterial prostatitis(CNP). Methods 80 cases of SD rats weredivided into blank group, model group and Oldenlandia extract low, middle and high dose group. Complete Freund's adjuvantinjection on ventral lobe of prostate gland method was used to construct CNP model. After 14 days of modeling, theintragastric administration of low, medium and high doses of Oldenlandia diffusa extract(75, 150 and 300 mg/kg,respectively) was started. The blank group was given the same amount of normal saline. After 3 weeks of administration, themorphology of the prostate and the number of white blood cells were observed. The levels of TLR4 and NF-κB in prostatetissues were detected by Western blot. Results The body weight of the model group was significantly lower than that of theblank group, and the difference was statistically significant(P<0.05). There was no significant difference between the weightof the intervention group and the blank group(P>0.05). The prostate wet weight and prostate wet weight/body weight ratio ofthe model group and the extract of Oldenlandia diffusa group were significantly higher than those of the blank group(P<0.05). The ratio gradually decreased with the increase of the concentration of the drug, and the difference was statisticallysignificant(P<0.05). The results of high power microscope showed that there was a statistically significant difference in thenumber of prostate leukocytes in each group(P=0.000). The results of Western blot showed that the expression of TLR4 andNF-κB p65 in prostate tissue decreased significantly after intervention with Oldenlandia diffusa extract(P<0.05), and thelevels of TLR4 and NF-κB p65 decreased gradually with the increase of drug concentration(P<0.05). Conclusion Oldenlandia diffusa extract could obviously alleviate the degree of prostate injury in CNP rats, and its mechanism might berelated to the inhibition of the TLR4/NF-κB pathway.
Objective To explore the influence of Oldenlandia diffusa extract on prostate toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)pathway of rats with chronic nonbacterial prostatitis(CNP). Methods 80 cases of SD rats weredivided into blank group, model group and Oldenlandia extract low, middle and high dose group. Complete Freund's adjuvantinjection on ventral lobe of prostate gland method was used to construct CNP model. After 14 days of modeling, theintragastric administration of low, medium and high doses of Oldenlandia diffusa extract(75, 150 and 300 mg/kg,respectively) was started. The blank group was given the same amount of normal saline. After 3 weeks of administration, themorphology of the prostate and the number of white blood cells were observed. The levels of TLR4 and NF-κB in prostatetissues were detected by Western blot. Results The body weight of the model group was significantly lower than that of theblank group, and the difference was statistically significant(P<0.05). There was no significant difference between the weightof the intervention group and the blank group(P>0.05). The prostate wet weight and prostate wet weight/body weight ratio ofthe model group and the extract of Oldenlandia diffusa group were significantly higher than those of the blank group(P<0.05). The ratio gradually decreased with the increase of the concentration of the drug, and the difference was statisticallysignificant(P<0.05). The results of high power microscope showed that there was a statistically significant difference in thenumber of prostate leukocytes in each group(P=0.000). The results of Western blot showed that the expression of TLR4 andNF-κB p65 in prostate tissue decreased significantly after intervention with Oldenlandia diffusa extract(P<0.05), and thelevels of TLR4 and NF-κB p65 decreased gradually with the increase of drug concentration(P<0.05). Conclusion Oldenlandia diffusa extract could obviously alleviate the degree of prostate injury in CNP rats, and its mechanism might berelated to the inhibition of the TLR4/NF-κB pathway.
引文
[1]齐曼芳,黄高翔,周雪娟.慢性非细菌性前列腺炎发病机制的研究进展[J].广西医科大学学报,2017,34(8):1247-1249.Qi MF,Huang GX,Zhou XJ.Research progress of the pathogenesis of chronic non-bacterial prostatitis[J].Journal of Guangxi Medical University,2017,34(8):1247-1249.
[2]赵丽杰,赵建宏.复方土茯苓片治疗前列腺炎的疗效及对细胞因子水平的影响[J].中国临床研究,2016,29(3):392-396.Zhao LJ,Zhao JH.Effect of compound Tufuling tablets on prostatitis and its effect on cytokines[J].Chinese Journal of Clinical Research,2016,29(3):392-396.
[3]Jin JX,Wang HZ,Zhai ZX,et al.Transrectal microwave thermotherapy causing a short-time influence on sperm quality in Chinese chronic nonbacterial prostatitis patients[J].Asian J Androl,2017,19(5):548-553.
[4]马河,李方丽,王芳,等.白花蛇舌草化学成分研究[J].中药材,2016,39(1):98-102.Ma H,Li FL,Wang F,et al.Chemical constituents from Oldenlandia diffusa[J].Journal of Chinese Medicinal Materials,2016,39(1):98-102.
[5]高红瑾,陆姗姗.白花蛇舌草乙醇提取物抗炎作用研究[J].中国现代药物应用,2017,11(15):195-196.Gao HJ,Lu SS.Study on the anti-inflammatory effect of ethanol extract of Hedyotis diffusa Willd[J].Chin J Mod Drug Appl,2017,11(15):195-196.
[6]Feng Y,Cui Y,Gao JL,et al.Resveratrol attenuates neuronal autophagy and inflammatory injury by inhibiting the TLR4/NF-κB signaling pathway in experimental traumatic brain injury[J].Int J Mol Med,2016,37(4):921-930.
[7]Ye HY,Jin J,Jin LW,et al.Chlorogenic acid attenuates lipopolysaccharide-induced acute kidney injury by inhibiting TLR4/NF-κB signal pathway[J].Inflammation,2017,40(2):523-529.
[8]宋国宏,李文玉,张晨,等.免疫法制作大鼠非细菌性前列腺炎模型的量效关系研究[J].中华男科学杂志,2011,17(7):586-590.Song GH,Li WY,Zhang C,et al.Establishment of a nonbacterial prostatitis model in rats using the autoimmune method:A dose-effect study[J].National Journal of Andrology,2011,17(7):586-590.
[9]尹学来,宋竖旗,刘冬,等.实验性自身免疫性前列腺炎疼痛大鼠模型的制作[J].北京医学,2016,38(3):253-256,289.Yin XL,Song SQ,Liu D,et al.The establishment of experimental autoimmune prostatitis rat model with pain[J].Beijing Medical Journal,2016,38(3):253-256,289.
[10]Zilberman DE,Tsivian M,Mor Y,et al.The trues behind TRUS in the setting of chronic prostatitis/chronic pelvic pain syndrome[J].Urological Science,2016,27(1):40-44.
[11]朱彩云,罗国胜,邱群芳,等.369例无症状男性前列腺液白细胞分析[J].热带医学杂志,2014,14(9):1183-1185.Zhu CY,Luo GS,Qiu QF,et al.369 cases of asymptomatic men with prostate fluid leukocyte analysis[J].J Trop Med,2014,14(9):1183-1185.
[12]马文军,胡青林,刁磊,等.药油灸对慢性非细菌性前列腺炎患者前列腺液中锌含量和C反应蛋白的影响[J].中国针灸,2017,37(8):840-844.Ma WJ,Hu QL,Diao L,et al.The effect of drug oil moxibustion for contents of Zinc and C-reactive protein in succus prostaticus of chronic nonbacterial prostatitis[J].Chinese Acupuncture&Moxibustion,2017,37(8):840-844.
[13]陈曦,莫林键,汪小明,等.不同NIH分型前列腺炎患者前列腺液白细胞及卵磷脂小体计数与症状指数的关系[J].广东医学,2015,36(16):2501-2504.Chen X,Mo LJ,Wang XM,et al.Analysis of correlation between quantity of leukocytes and small particles of lecithin and the National Institutes of Health chronic prostatitis symptom index in different NIH categories of prostatitis[J].Guangdong Medical Journal,2015,36(16):2501-2504.
[14]Elshabrawy HA,Essani AE,Szekanecz Z,et al.TLRs,future potential therapeutic targets for RA[J].Autoimmun Rev,2017,16(2):103-113.
[15]Behzadi E,Behzadi P.The role of toll-like receptors(TLRs)in urinary tract infections(UTIs)[J].Cent European J Urol,2016,69(4):404-410.
[16]Dong Z,Xiong L,Zhang W,et al.Holding the inflammatory system in check:TLRs and their targeted therapy in asthma[J].Mediators Inflamm,2016,2016:2180417.
[17]李芳,杨培民,曹广尚.白花蛇舌草黄酮类成分大鼠在体肠吸收研究[J].中草药,2015,46(2):240-244.Li F,Yang PM,Cao GS.In situ intestinal absorption of flavonoids from Hedyotis diffusa in rats[J].Chinese Traditional and Herbal Drugs,2015,46(2):240-244.
[18]张林超,赵俊峰,孙继建,等.白花蛇舌草乙醇提取物对人肾癌细胞786-0体外增殖及小鼠肾癌细胞786-0移植瘤生长的影响[J].山东医药,2017,57(23):20-23.Zhang LC,Zhao JF,Sun JJ,et al.Effects of ethanol extracts of Hedyotis diffusae herba on in-vitro proliferation of human renal cancer 786-0 cells and growth of mouse renal cancer 786-0 cell tumor xenograft[J].Shandong Medical Journal,2017,57(23):20-23.
[19]聂利华,廖鹏,刘亚群.白花蛇舌草醇提物抗氧化活性的研究[J].中南药学,2017,15(1):44-47.Nie LH,Liao P,Liu YQ.Antioxidant activity of ethanol extract from Hedyotis diffusa Willd[J].Central South Pharmacy,2017,15(1):44-47.
[20]罗世英,周乐,吕小华,等.白花蛇舌草总黄酮对实验性溃疡性结肠炎的作用及免疫学机制研究[J].中国中药杂志,2014,39(5):896-900.Luo SY,Zhou L,LüXH.Study on effect of total flavonoids of Oldenlendia difflusa on ulcerative colitis and its immunological mechanism[J].China Journal of Chinese Materia Medica,2014,39(5):896-900.
[2]赵丽杰,赵建宏.复方土茯苓片治疗前列腺炎的疗效及对细胞因子水平的影响[J].中国临床研究,2016,29(3):392-396.Zhao LJ,Zhao JH.Effect of compound Tufuling tablets on prostatitis and its effect on cytokines[J].Chinese Journal of Clinical Research,2016,29(3):392-396.
[3]Jin JX,Wang HZ,Zhai ZX,et al.Transrectal microwave thermotherapy causing a short-time influence on sperm quality in Chinese chronic nonbacterial prostatitis patients[J].Asian J Androl,2017,19(5):548-553.
[4]马河,李方丽,王芳,等.白花蛇舌草化学成分研究[J].中药材,2016,39(1):98-102.Ma H,Li FL,Wang F,et al.Chemical constituents from Oldenlandia diffusa[J].Journal of Chinese Medicinal Materials,2016,39(1):98-102.
[5]高红瑾,陆姗姗.白花蛇舌草乙醇提取物抗炎作用研究[J].中国现代药物应用,2017,11(15):195-196.Gao HJ,Lu SS.Study on the anti-inflammatory effect of ethanol extract of Hedyotis diffusa Willd[J].Chin J Mod Drug Appl,2017,11(15):195-196.
[6]Feng Y,Cui Y,Gao JL,et al.Resveratrol attenuates neuronal autophagy and inflammatory injury by inhibiting the TLR4/NF-κB signaling pathway in experimental traumatic brain injury[J].Int J Mol Med,2016,37(4):921-930.
[7]Ye HY,Jin J,Jin LW,et al.Chlorogenic acid attenuates lipopolysaccharide-induced acute kidney injury by inhibiting TLR4/NF-κB signal pathway[J].Inflammation,2017,40(2):523-529.
[8]宋国宏,李文玉,张晨,等.免疫法制作大鼠非细菌性前列腺炎模型的量效关系研究[J].中华男科学杂志,2011,17(7):586-590.Song GH,Li WY,Zhang C,et al.Establishment of a nonbacterial prostatitis model in rats using the autoimmune method:A dose-effect study[J].National Journal of Andrology,2011,17(7):586-590.
[9]尹学来,宋竖旗,刘冬,等.实验性自身免疫性前列腺炎疼痛大鼠模型的制作[J].北京医学,2016,38(3):253-256,289.Yin XL,Song SQ,Liu D,et al.The establishment of experimental autoimmune prostatitis rat model with pain[J].Beijing Medical Journal,2016,38(3):253-256,289.
[10]Zilberman DE,Tsivian M,Mor Y,et al.The trues behind TRUS in the setting of chronic prostatitis/chronic pelvic pain syndrome[J].Urological Science,2016,27(1):40-44.
[11]朱彩云,罗国胜,邱群芳,等.369例无症状男性前列腺液白细胞分析[J].热带医学杂志,2014,14(9):1183-1185.Zhu CY,Luo GS,Qiu QF,et al.369 cases of asymptomatic men with prostate fluid leukocyte analysis[J].J Trop Med,2014,14(9):1183-1185.
[12]马文军,胡青林,刁磊,等.药油灸对慢性非细菌性前列腺炎患者前列腺液中锌含量和C反应蛋白的影响[J].中国针灸,2017,37(8):840-844.Ma WJ,Hu QL,Diao L,et al.The effect of drug oil moxibustion for contents of Zinc and C-reactive protein in succus prostaticus of chronic nonbacterial prostatitis[J].Chinese Acupuncture&Moxibustion,2017,37(8):840-844.
[13]陈曦,莫林键,汪小明,等.不同NIH分型前列腺炎患者前列腺液白细胞及卵磷脂小体计数与症状指数的关系[J].广东医学,2015,36(16):2501-2504.Chen X,Mo LJ,Wang XM,et al.Analysis of correlation between quantity of leukocytes and small particles of lecithin and the National Institutes of Health chronic prostatitis symptom index in different NIH categories of prostatitis[J].Guangdong Medical Journal,2015,36(16):2501-2504.
[14]Elshabrawy HA,Essani AE,Szekanecz Z,et al.TLRs,future potential therapeutic targets for RA[J].Autoimmun Rev,2017,16(2):103-113.
[15]Behzadi E,Behzadi P.The role of toll-like receptors(TLRs)in urinary tract infections(UTIs)[J].Cent European J Urol,2016,69(4):404-410.
[16]Dong Z,Xiong L,Zhang W,et al.Holding the inflammatory system in check:TLRs and their targeted therapy in asthma[J].Mediators Inflamm,2016,2016:2180417.
[17]李芳,杨培民,曹广尚.白花蛇舌草黄酮类成分大鼠在体肠吸收研究[J].中草药,2015,46(2):240-244.Li F,Yang PM,Cao GS.In situ intestinal absorption of flavonoids from Hedyotis diffusa in rats[J].Chinese Traditional and Herbal Drugs,2015,46(2):240-244.
[18]张林超,赵俊峰,孙继建,等.白花蛇舌草乙醇提取物对人肾癌细胞786-0体外增殖及小鼠肾癌细胞786-0移植瘤生长的影响[J].山东医药,2017,57(23):20-23.Zhang LC,Zhao JF,Sun JJ,et al.Effects of ethanol extracts of Hedyotis diffusae herba on in-vitro proliferation of human renal cancer 786-0 cells and growth of mouse renal cancer 786-0 cell tumor xenograft[J].Shandong Medical Journal,2017,57(23):20-23.
[19]聂利华,廖鹏,刘亚群.白花蛇舌草醇提物抗氧化活性的研究[J].中南药学,2017,15(1):44-47.Nie LH,Liao P,Liu YQ.Antioxidant activity of ethanol extract from Hedyotis diffusa Willd[J].Central South Pharmacy,2017,15(1):44-47.
[20]罗世英,周乐,吕小华,等.白花蛇舌草总黄酮对实验性溃疡性结肠炎的作用及免疫学机制研究[J].中国中药杂志,2014,39(5):896-900.Luo SY,Zhou L,LüXH.Study on effect of total flavonoids of Oldenlendia difflusa on ulcerative colitis and its immunological mechanism[J].China Journal of Chinese Materia Medica,2014,39(5):896-900.