Influences of Toll-like Receptor-4 Gene Polymorphism on Occurance of Stroke Due to Qi Deficiency and Blood Stasis in Han Chinese Men

OBJECTIVE To explore the correlation between polymorphisms in the toll-like receptor 4 (rs1927914， rs10759932 and rs11536889) and susceptivity， gene expression， cytokines and blood lipid metabolism in southern Han Chinese with stroke due to wind-phlegm obstruction， qi deficiency and blood stasis. METHODS 559 stroke cases were divided into two groups based on case-control study method， with 248 cases in the qi deficiency and blood stasis group， 311 cases in wind-phlegm obstruction group. In addition， another 605 cases were involved as the healthy control group with the corresponding gender and age. All the subjects were from the Han nationality. Sequenom MassARRAY technology was adopted for testing and genotyping of rs1927914， rs10759932 and rs11536889 in the toll-like receptor 4. Real-time fluorescence quantification(qPCR) was applied for examine mRNA expression. Enzyme linked immunosorbent assay(ELISA) was used to detect the level of interleukin-1β(IL-1β)， interleukin-6 (IL-6)， interleukin-8 (IL-8)， interleukin-12(IL-12) and tumor necrosis factor-α(TNF-α). HITACHI 7600 automatic biochemical analyzer was applied to test the total cholesterol(TC)， triglyceride(TG)， high density lipoprotein cholesterol(HDL) and low density lipoprotein cholesterol(LDL). RESULTS There existed a statistical difference between the male cases of the qi deficiency and blood stasis group and male cases of the control group in the distribution of gene frequency of rs10759932(P<0.05). The locus polymorphism of both rs1927914 (dominant model: OR=0.65， 95%CI=0.43-0.96， Padj=0.031) and rs10759932 (dominant model: OR=0.64， 95%CI=0.41-0.99， Padj=0.046) were closely related with susceptivity of stroke male of qi deficiency and blood stasis type. The IL-8 level of male stroke patients of qi deficiency and blood stasis type with the rs10759932 TT genotype was much lower than those with CC+CT(P<0.01). Rs11536889 polymorphism was notablely related to TG level(additive model: Padj=0.021， dominant model: Padj=0.013) and HDL level(dominant model: Padj=0.032) in stroke patients of qi deficiency and blood stasis type. CONCLUSION TLR4 polymorphisms exert certain influence on the occurance of stroke due to qi deficiency and blood stasis in Han Chinese men and they have been involved in the inflammatory reaction and blood lipid metabolism.