雷公藤多苷对脂多糖诱导大鼠星形胶质细胞产生炎症因子的影响

Effect of tripterygium glycosides on inflammatory factors induced by lipopolysaccharide in rat astrocytes

目的观察雷公藤多苷对脂多糖诱导的大鼠星形胶质细胞炎症反应的影响。方法取新生3 d内SD大鼠的大脑皮质,分离提取星形胶质细胞,用脂多糖1μg·mL^(-1)刺激星形胶质细胞制备炎症模型。将分离纯化后的星形胶质细胞细胞分为5组:空白组、模型组和低、中、高3个剂量实验组,空白组加基础培养基;模型组加1μg·mL^(-1)脂多糖;低、中、高3个剂量实验组分别加入25,50,100μg·L^(-1)雷公藤多苷预处理细胞,然后加入1μg·mL^(-1)脂多糖诱导细胞。以CCK8检测各组细胞存活率,以酶联免疫吸附实验检测肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)的表达变化,蛋白质免疫印迹法分析细胞中一氧化氮合酶(i NOS)蛋白表达变化。结果空白组、模型组和低、中、高3个剂量实验组的细胞存活率分别为(100. 33±5. 61)%,(50. 74±6. 67)%,(74. 85±3. 48)%,(85. 68±5. 51)%,(94. 12±3. 43)%;这5组的TNF-α(pg·mL^(-1))分别为39. 23±15. 34,169. 43±15. 19,117. 83±12. 16,80. 81±5. 28,57. 87±7. 92;这5组的IL-6 (pg·mL^(-1))分别为47. 62±7. 93,299. 46±18. 45,232. 57±23. 18,170. 66±26. 67,94. 51±24. 36;这5组的i NOS表达灰度值分别为0. 32±0. 02,1. 10±0. 04,0. 78±0. 05,0. 58±0. 04,0. 37±0. 04,模型组与空白组比较,上述指标差异均有统计学意义(均P <0. 05);低、中、高3个剂量实验组与模型组比较,上述指标差异均有统计学意义(P <0. 05,P <0. 001)。结论雷公藤多苷可明显改善脂多糖诱导的星形胶质细胞炎症损伤。

Objective To observe the effects of tripterygium glycosides on lipopolysaccharide-induced inflammatory response in astrocyte.Methods The astrocytes were isolated and extracted from cerebral cortex of newborn 3 d SD rats,astrocyte inflammatory model was prepared by 1 μg·mL^-1 lipopolysaccharide. The isolated and purified astrocytes were divided into 5 groups: blank group,model group,and experimental-L,experimental-M,experimental-H groups,basic medium was added in the blank group;1 μg · mL^-1 lipopolysaccharide was added in the model group;25,50,100 μg · L^-1 tripterygium glycosides were respectively added in experimental-L,-M,-H groups,then add 1 μg·m L^-1 lipopolysaccharide induces cells. CCK8 was used to detect the survival rate of each group. The expression of tumor necrosis factor-α( TNF-α) andinterleukin-6( IL-6) were detected by enzyme-linked immunosorbent assay. The inducible nitric oxide synthase( iNOS) protein was analyzed by Western blotting. Results The cell survival rates in blank group,model group,and low,medium,high dose experimental groups were( 100. 33 ± 5. 61) %,( 50. 74 ± 6. 67) %,( 74. 85 ± 3. 48) %,( 85. 68 ± 5. 51) %,( 94. 12 ± 3. 43) %. After pretreatment of astrocytes with tripterygium glycosides,the TNF-α( pg·mL-1) in the 5 groups were 39. 23 ± 15. 34,169. 43 ± 15. 19,117. 83 ± 12. 16,80. 81 ± 5. 28,57. 87 ± 7. 92;the IL-6( pg · mL^-1) in the 5 groups were 47. 62 ± 7. 93,299. 46 ± 18. 45,232. 57 ± 23. 18,170. 66 ± 26. 67,94. 51 ± 24. 36;the gray values of i NOS expression in 5 groups were 0. 32 ± 0. 02,1. 10 ± 0. 04,0. 78 ± 0. 05,0. 58 ± 0. 04,0. 37 ± 0. 04. Comparison between model group and blank group,the difference of all factors were statistically significant( all P < 0. 05). Comparison between the low,medium and high doses of the experimental groups and model group,the difference of all factors were statistically significant( P < 0. 05,P < 0. 001). Conclusion Tripterygium glycosides can significantly improve lipopolysaccharide induced astrocyte inflammatory injury.