Th1/Th2平衡对寒凝血瘀证原发性痛经大鼠神经生长因子及其受体的影响

Th1/Th2 Balance Affects Nerve Growth Factor and Its Receptors in Primary Dysmenorrhea Rats with Syndrome of Cold Coagulation and Blood Stasis

目的 探究1型辅助性T细胞/2型辅助性T细胞(Type 1 helper T cells/type 2 helper T cells, Th1/Th2)平衡偏移对寒凝血瘀证原发性痛经(Primary dysmenorrhea, PD)大鼠子宫组织神经生长因子(Nerve growth factor, NGF)及其受体的影响,进一步揭示PD的发病机制。方法 将32只雌性大鼠,按随机数字表法分为空白组、模型组、Th1偏移组、Th2偏移组,每组各8只。建立寒凝血瘀证PD大鼠模型,在此模型基础上建立Th1偏移PD模型及Th2偏移PD模型。比较各组大鼠扭体反应,检测各组大鼠子宫组织中神经生长因子(Nerve growth factor, NGF)、酪氨酸激酶受体A(Tropomyosin-receptor kinase, TrkA)及p75营养因子受体(p75 neurotrophin receptor, p75NTR)蛋白含量及mRNA转录水平。结果 扭体潜伏时间:与模型组比较,Th1偏移组和Th2偏移组,差异无统计学意义(P>0.05);Th2偏移组较Th1偏移组缩短,差异有统计学意义(P<0.05)。30 min内扭体次数:与模型组比较,Th1偏移组明显减少,差异有统计学意义(P<0.01),Th2偏移组明显增加,差异有统计学意义(P<0.01)。与空白组比较,模型组大鼠NGF、TrkA及p75NTR蛋白表达及mRNA水平均升高,差异有统计学意义(P<0.05,P<0.01);与模型组比较,Th1偏移组NGF、TrkA蛋白表达及mRNA水平降低,差异有统计学意义(P<0.05,P<0.01),p75NTR mRNA水平降低,差异有统计学意义(P<0.01);Th2偏移组NGF、TrkA及p75NTR蛋白表达及mRNA水平均升高,差异有统计学意义(P<0.05,P<0.01)。结论 Th1/Th2细胞平衡干预NGF及其受体表达可能是寒凝血瘀证PD的发病机制之一。

Objective To observe the effects of type 1 helper T cells/type 2 helper T cells(Th1/Th2)balance drift on nerve growth factor(NGF)and its receptors in primary dysmenorrhea(PD)rats with the syndrome of cold coagulation and blood stasis and further reveal the pathogenesis of PD.Methods The rat model of PD with the syndrome of cold coagulation and blood stasis was established.On the basis of this model,Th1 shifting PD model and Th2 shifting PD model were established.The rats were randomized into normal,model,Th1 shifting,and Th2 shifting groups according to the random number table method.The writhing latency and the writhing times in 30 min were recorded.The protein and mRNA levels of NGF,tropomyosin-receptor kinase(TrkA),and p75 neurotrophin receptor(p75NTR)in the uterine tissue were determined by western blotting and real-time PCR,respectively.Results The model group had no significant difference in the writhing latency from Th1 shifting group and Th2 shifting group(P>0.05),and the Th2 shifting group had shorter writhing latency than Th1 shifting group(P<0.05).Compared with the model group,the Th1 shifting group showed decreased times of writhing within 30 min(P<0.01),while the Th2 shifting group showed increased times of writhing within 30 min(P<0.01).Compared with the normal group,the model group showed up-regulated protein and mRNA levels of NGF,TrkA,and p75NTR(P<0.05,P<0.01).Compared with the model group,the Th1 shifting group showed down-regulated protein and mRNA levels of NGF and TrkA(P<0.05,P<0.01)and down-regulated mRNA level of p75NTR(P<0.01);the Th2 shifting group showed up-regulated protein and mRNA levels of NGF,TrkA,and p75NTR(P<0.05,P<0.01).Conclusion Th1/Th2 balance shift disturbing the expression of NGF and its receptors may be one of the pathogenesis of PD with the syndrome of cold coagulation and blood stasis.