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目的:检测湿热消腰部热敷疗法对实验性自身免疫性前列腺炎(EAP)小鼠模型前列腺组织Th17/Treg相关特异性因子表达的影响,探讨湿热消腰部热敷疗法可能的作用机制。方法:120只健康雄性KM小鼠,随机抽取20只为正常组,其余100只应用Wistar大鼠前列腺蛋白提纯液辅以免疫佐剂及中医病因造模法制备实验性自身免疫性前列腺炎(湿热证)小鼠模型,造模成功后的小鼠随机分为模型组、基质组、低剂量组、中剂量组和高剂量组。脱去各组小鼠腰1~腰3椎体处毛发,低剂量组、中剂量组和高剂量组用湿热消棉垫、基质组用凡士林棉垫、正常组及模型组用生理盐水棉垫,所用棉垫加热至45℃左右外敷于小鼠脱毛区,每次治疗10 min,正常组、模型组、基质组和低剂量组1次/d,中剂量组2次/d,高剂量组3次/d,连续干预4周后处死,用蛋白免疫印迹和实时荧光定量PCR检测各组小鼠前列腺组织中的视黄酸相关核孤儿受体γt(RORγt)和双头叉转录因子p3(Foxp3)基因蛋白及其mRNA表达。结果:造模后,模型小鼠的症状、体征及病理学改变基本符合慢性前列腺炎(湿热证)的表现。干预后,模型组与正常组相比,前列腺中Foxp3及其mRNA表达明显下降,RORγt及其mRNA表达显著提高,差异有统计学意义(P<0.05);与模型组比较,中、高剂量组RORγt及其mRNA的表达显著降低(P<0.05),高剂量组Foxp3的表达显著提高(P<0.05),低、中、高剂量组Foxp3 mRNA的表达均无明显改变(P>0.05)。结论:湿热消腰部热敷疗法可能是通过下调RORγt基因的表达,抑制Th17细胞的分化,使Th17/Treg分化失衡得到控制而在慢性前列腺炎的治疗中发挥作用。
OBJECTIVE: To investigate the effect of dampness and heat-dissipating waist heat therapy on the expression of Th17 / Treg-related specific factors in prostatic tissue of experimental autoimmune prostatitis (EAP) mouse model, and to explore the possible mechanism of damp-heat elimination waist heat therapy. Methods: A total of 120 healthy male KM mice were randomly selected from 20 normal rats. The remaining 100 rabbits were used to prepare experimental autoimmune prostatitis (moist heat Mice). After successful model establishment, the mice were randomly divided into model group, matrix group, low dose group, middle dose group and high dose group. Remove the mice lumbar 1 ~ lumbar 3 vertebral hair, low-dose group, the middle-dose group and high-dose group with wet heat pad, matrix group with Vaseline pad, the normal group and model group with saline pad The cotton pad was heated to about 45 ℃ and applied to the hair removal area of mice for 10 minutes. The normal group, the model group, the matrix group and the low dose group were given once / d, the middle dose group were given twice a day, the high dose group 3 times / d, and were sacrificed after 4 weeks of continuous intervention. The expression of retinoic acid-related nuclear orphan receptor γt (RORγt) and double-headed fork transcription factor p3 (p38) in the prostatic tissues were detected by Western blotting and real- Foxp3) gene protein and its mRNA expression. Results: After modeling, the symptoms, signs and pathological changes in model mice were basically consistent with those of chronic prostatitis (damp-heat syndrome). Compared with the normal group, the expression of Foxp3 and its mRNA in the model group and the mRNA expression of RORγt in the model group were significantly increased after intervention (P <0.05). Compared with the model group, The mRNA expression of RORγt and its mRNA were significantly decreased (P <0.05). The expression of Foxp3 in high dose group was significantly increased (P <0.05), and the expression of Foxp3 mRNA in low dose, middle dose and high dose group had no significant change (P> 0.05). CONCLUSION: The hot compress therapy of lumbar heat and dampness may play a role in the treatment of chronic prostatitis by down-regulating the expression of RORγt gene, inhibiting the differentiation of Th17 cells and controlling the imbalance of Th17 / Treg differentiation.