补肾温阳化瘀汤对子宫内膜异位症痛经小鼠模型中神经生长因子及缓激肽/缓激肽B1受体的影响

Effect of Bushenwenyanghuayu decoction on nerve growth factor and bradykinin/bradykinin B1 receptor in a endometriosis dysmenorrhea mouse model.

作者信息

Jingwei Chen, Huilan Du, Ruixiao Tong, Hua Yang, Huirong Ma

出版信息

J Tradit Chin Med. 2015 Apr;35(2):184-91. doi: 10.1016/s0254-6272(15)30026-1.

DOI:10.1016/s0254-6272(15)30026-1

PMID:25975051

Abstract

OBJECTIVE

To observe the effects of Bushenwenyanghuayu decoction (BD), a Traditional Chinese Medicine (TCM), on the serum concentration of nerve growth factor (NGF) and bradykinin (BK), and protein and mRNA levels of NGF and bradykinin B1 receptor (BKB1R) in a mouse model of endometriosis dysmenorrhea.

METHODS

Seventy-five experimental female BALB/c mice were randomly divided into five groups, 15 mice each: sham, model, BD high dose (61.67 g/kg), BD low dose (15.42 g/kg), and gestrinone (0.4 mg/kg) groups. All the mice except for those in the sham group underwent auto-transplantation surgery and were gavaged estradiol valerate (0.5 mg/kg, daily for 12 days) after surgery. On the 12th day, 1 h after administration, writhing response was induced by intraperitoneal injection of oxytocin at 2 U/mouse. The writhing frequency and latency were recorded and the volume of the ectopic foci was measured. The concentration of serum NGF and BK was detected by enzyme-linked immunosorbent assay, the protein expression of NGF and BKB1R was tested by immunohistochemistry and western blotting, and NGF and BKB1R mRNAs were detected by real-time PCR.

RESULTS

Compared with the model group, the volume of the ectopic foci in the treatment groups was significantly lower (P < 0.01), the writhing frequency was decreased (P < 0.05), and the writhing latency was prolonged (P < 0.01). Compared with the sham group, serum NGF and BK levels in the model group were significantly increased (P < 0.01). There were positive correlations for writhing frequency among the NGF and BK groups (P < 0.01). The serum NGF and BK levels were significantly lower in the treatment groups than the model group (P < 0.05). The protein expression of NGF, BKB1R was significantly decreased in the treatment groups compared with the model group (P < 0.01). NGF and BKB1R mRNA expression was significantly decreased in the treatment groups compared with the model group (P < 0.01).

CONCLUSION

NGF and BK/BKB1R may play an important role in the development of endometriosis-associated dysmenorrhea, and BD was found to inhibit the development of endometriosis and relieve dysmenorrhea by influencing NGF and BK/ BKB1R mRNA and protein levels.

摘要

目的

观察中药补肾温阳化瘀汤（BD）对子宫内膜异位症痛经小鼠模型血清神经生长因子（NGF）和缓激肽（BK）浓度，以及NGF和缓激肽B1受体（BKB1R）蛋白及mRNA水平的影响。

方法

将75只实验雌性BALB/c小鼠随机分为5组，每组15只：假手术组、模型组、BD高剂量组（61.67 g/kg）、BD低剂量组（15.42 g/kg）和孕三烯酮组（0.4 mg/kg）。除假手术组小鼠外，其余小鼠均行自体移植手术，术后给予戊酸雌二醇灌胃（0.5 mg/kg，每日1次，共12天）。第12天给药1小时后，腹腔注射缩宫素2 U/只诱发扭体反应。记录扭体频率和潜伏期，并测量异位病灶体积。采用酶联免疫吸附测定法检测血清NGF和BK浓度，采用免疫组织化学和蛋白质印迹法检测NGF和BKB1R蛋白表达，采用实时荧光定量PCR检测NGF和BKB1R mRNA。

结果

与模型组相比，各治疗组异位病灶体积显著减小（P < 0.01），扭体频率降低（P < 0.05），扭体潜伏期延长（P < 0.01）。与假手术组相比，模型组血清NGF和BK水平显著升高（P < 0.01）。NGF和BK组扭体频率之间存在正相关（P < 0.01）。各治疗组血清NGF和BK水平均显著低于模型组（P < 0.05）。与模型组相比，各治疗组NGF、BKB1R蛋白表达显著降低（P < 0.01）。与模型组相比，各治疗组NGF和BKB1R mRNA表达显著降低（P < 0.01）。