补肾填精法对肾精亏虚孕鼠脾脏及TLR4/MyD88/NF-κBp65免疫通路的影响

Effect of Tonifying Kidney and Replenishing Essence on the Spleen and the TLR4/MyD88/NF-κBp65 Immune Pathway in Pregnant Rats with Kidney Essence Deficiency Syndrome

目的探讨补肾填精法对肾精亏虚孕鼠的脾脏及其相关免疫通路的影响。方法选用复合应激法建立肾精亏虚孕鼠模型,并给予左归丸混悬液(0.58 g/100 g·d^-1)灌胃处理以观察补肾填精法对模型孕鼠脾脏显微和超微结构变化及其相关免疫通路中的Toll样受体4(Toll-like receptor 4,TLR4)、髓样分化因子88(Myeloid differentiation factor 88,MyD88)和核转录因子κBp65(Nuclear factor-kappa beta p65,NF-κBp65)的表达水平的影响。结果正常组脾脏结构完整,无明显的细胞凋亡和坏死现象,细胞内结构正常。模型组脾脏结构稀疏,淋巴小结缩小且界限模糊,淋巴细胞数目减少,有明显的细胞凋亡和坏死现象;细胞内核膜不规则,染色质有固缩现象,部分细胞器崩解。补肾组脾脏结构明显改善,细胞凋亡和坏死现象减少,细胞内各结构的形态显著恢复。此外,与正常组比较,模型组孕鼠脾脏内TLR4、MyD88和NF-κBp65的mRNA和蛋白质表达水平下降。与模型组比较,补肾组中上述三个因子的mRNA和蛋白质表达水平明显回升。结论补肾填精法可以通过改善脾脏结构和上调TLR4/MyD88/NF-κBp65免疫通路的表达水平来有效地调节孕鼠的免疫失衡状态,有利于治疗肾精亏虚引起的胎漏、胎动欲坠等症。

Objective To explore the effect of tonifying kidney and replenishing essence on the spleen and its related immune pathway in pregnant rats with kidney essence deficiency syndrome.Methods The pregnant rat model of kidney essence deficiency syndrome was established by compound stress method and treated intragastrically with Zuogui pill suspension(0.58 g/100 g·d^-1)to observe the effect of tonifying kidney and replenishing essence on the changes of spleen microstructure and ultrastructure and on the expression levels of toll-like receptor 4(TLR4),myeloid differentiation factor 88(MyD88)and nuclear factor-kappa beta p65(NF-κBp65)in its related immune pathway.Results In normal group,the spleen structure was intact,no obvious apoptosis and necrosis was observed,and the intracellular structure was normal.In model group,the spleen structure was sparse,the area of lymphoid nodules was reduced and the boundary was blurred.Also,the number of lymphocytes decreased,with obvious apoptosis and necrosis.Besides,the intracellular nuclear membrane was irregular,the chromatin had condensation,and some organelles collapsed.In kidney-tonifying group,the spleen structure was significantly improved,the apoptosis and necrosis decreased significantly and the morphology of intracellular structure was obviously restored.Further results showed that the mRNA and protein expression levels of TLR4,MyD88 and NF-κBp65 in the spleens of pregnant rats were significantly decreased in the model group compared with the normal group.And the expression levels of these three factors in the spleens of pregnant rats recovered significantly in the kidney-tonifying group compared with the model group.Conclusion Tonifying kidney and replenishing essence could effectively regulate the immune imbalance in pregnant rats with kidney essence deficiency syndrome by improving the structure of the spleen and increasing the expression levels of TLR4/MyD88/NF-κBp65 immune pathway.It would be beneficial for guiding treatment of abortion,embryo instability caused by deficiency of kidney essence.