CAJ | 학술논문

目的探讨凝血酶对培养的大鼠脑星形胶质细胞毒性损害及水蛭提取液对其的保护作用. 方法建立体外实验即大鼠脑星形胶质细胞培养实验模型,相差显微镜观察细胞生长状况.MTT法筛选水蛭提取液的有效浓度.星形胶质细胞分为水蛭提取液治疗组与凝血酶对照组,测定培养上清液酸脱氢酶(LDH)活性(细胞死亡的标志).免疫组化法观察它们的HSP70和TGFβ-1的表达. 结果 (1)一定浓度范同(1～100 U/mL)的凝血酶对星形胶质细胞有毒性作用,且随凝血酶剂量的增大,它对星形胶质细胞的毒性作用相应增加(F=118.65,P=0.000).(2)一定浓度范围内(0.25～4 mg/μL)的水蛭提取液能明显减轻10 U/mL凝血酶对星形胶质细胞的毒性作用(F=156.08,P=0.000),且随水蛭提取液浓度的增大,保护作用增强,还可促进星形胶质细胞HSP70和TGFβ-1的表达. 结论水蛭提取液能促进星形胶质细胞增生,增强它们的HSP70和TGFβ-1表达,从而明显抑制凝血酶对星形胶质细胞的毒性作用.
목적 탐토응혈매대배양적대서뇌성형효질세포독성손해급수질제취액대기적보호작용. 방법 건입체외실험즉대서뇌성형효질세포배양실험모형,상차현미경관찰세포생장상황.MTT법사선수질제취액적유효농도.성형효질세포분위수질제취액치료조여응혈매대조조,측정배양상청액산탈경매(LDH)활성(세포사망적표지).면역조화법관찰타문적HSP70화TGFβ-1적표체. 결과 (1)일정농도범동(1～100 U/mL)적응혈매대성형효질세포유독성작용,차수응혈매제량적증대,타대성형효질세포적독성작용상응증가(F=118.65,P=0.000).(2)일정농도범위내(0.25～4 mg/μL)적수질제취액능명현감경10 U/mL응혈매대성형효질세포적독성작용(F=156.08,P=0.000),차수수질제취액농도적증대,보호작용증강,환가촉진성형효질세포HSP70화TGFβ-1적표체. 결론 수질제취액능촉진성형효질세포증생,증강타문적HSP70화TGFβ-1표체,종이명현억제응혈매대성형효질세포적독성작용.
Objective To study the cell toxicity of thrombin in astrocytes in vitro and the protective effect of hirudo extract liquid (HEL) on the injured astrocytes. Methods Astrocytes were isolated from Wistar rats' cerebral cortex and cultured in vitro, and observed under a phase contrast microscope for growth status. Cell activity was measured with MTT assay. The survival of astrocytes was investigated after exposed to a selected concentration of thrombin ranging from 0.1 to 100 U/mL or to HEL ranging from 0.25 to 4 mg/μL by observing cell morphology under an inverted phase-contrast microscope and measuring the lactate dehydrogenase (LDH) activity (a marker of cell death) in cell supernatant. Expressions of HSP70 and TGFβ-1 protein in astrocytes were investigated by immunohistochemistry. Results (1) Thrombin (1-100 U/mL) had toxicity on astrocytes in vitro in a dose-dependent manner (F=118.65, P=0.000). (2) HEL (0.25-4 mg/μL) could significantly reduce the cell toxicity of 10 U/mL thrombin in astrocytes (F=156.08, P=0.000). With the increasing concentration of HEL, the protection of HEL was accordingly enhanced, and it even increased the expressions of HSP70and TGFβ-1. Conclusions HEL could accelerate the proliferation of astrocytes, enhance the expressions of HSP70 and TGFβ-1 protein, so as to significantly depress the cell toxicity of thrombin to astrocytes.