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目的检测门静脉高压症(PH)脾亢脾和正常脾巨噬细胞(Mφ)中Toll样受体2、4(TLR2、4)mRNA的表达差异,为进一步深入探讨Toll样受体在PH脾亢发生中的作用奠定基础。方法选取门静脉高压症脾亢患者(均为慢性乙型肝炎患者)的手术切除脾脏(12例)为实验组,外伤性脾破裂患者的手术切除脾脏(4例)为正常对照组。贴壁培养法分离纯化脾脏组织Mφ,荧光定量PCR法对Mφ表面Toll样受体2、4mRNA的表达进行检测,并将两组结果进行统计学分析比较。结果与正常脾脏相比,PH脾亢脾MφTLR2、4mRNA的表达水平明显增强(TLR2:2.29±0.55vs1.06±0.53,P<0.05;TLR4:2.32±0.41vs1.01±0.14,P<0.01)。结论PH脾亢脾MφTLR2、4的mRNA表达水平明显升高,与蛋白水平免疫组化的结果一致,进一步支持了“内毒素血症→脾脏MφToll样受体活化→Mφ吞噬破坏血细胞增多”是PH脾亢发生可能机制的观点。
Objective To detect the difference of Toll-like receptor 2 and 4 (TLR2,4) mRNA expression in portal hypertension (PH) spleen hypertrophic spleen and normal splenic macrophage (Mφ). To further explore the role of Toll- The role of occurred in laying the foundation. Methods 12 cases of splenectomy (12 cases) with portal hypertensive hypersplenism (all patients with chronic hepatitis B) were selected as the experimental group. Surgical resection of the spleen (4 cases) was performed as the normal control group. Mφ was isolated and purified from spleen tissue by adherent culture method. The expression of Toll-like receptor 2 and 4 mRNA on Mφ surface was detected by fluorescence quantitative PCR, and the results of two groups were statistically analyzed and compared. Results Compared with the normal spleen, the expression of MTLTL2.4mRNA in PH hypersplenism spleen was significantly increased (TLR2: 2.29 ± 0.55vs1.06 ± 0.53, P <0.05; TLR4: 2.32 ± 0.41vs1.01 ± 0.14, P <0.01) . Conclusions The mRNA expression level of TLR2 and TLR2 in splenic hypersplenism patients is obviously increased, which is consistent with the results of protein immunohistochemistry. It further supports “endotoxemia → activation of splenic MφToll-like receptors → phagocytosis of Mφ-induced cytocytosis” PH is the possible mechanism of hypersplenism point of view.