摘要
目的:观察姜黄素对脂多糖诱导的星形胶质细胞和小胶质细胞中角质细胞源性趋化因子(CXCL1)和干扰素诱导蛋白10(CXCL10)表达的抑制作用。方法:(1)确定脂多糖刺激细胞的时间:将培养的C6星形胶质细胞株随机分为对照组、脂多糖1 h、3 h、6 h组。对照组未加入任何药物刺激;脂多糖1 h、3 h、6 h组应用1μg/mL脂多糖分别刺激细胞1 h,3 h,6 h。采用real-time PCR方法检测细胞中趋化因子CXCL1和CXCL10 mRNA的表达,确定合适的脂多糖刺激时间。(2)姜黄素处理细胞实验:①C6细胞随机分成5组:对照组、脂多糖组、姜黄素2.5μmol/L、10μmol/L、25μmol/L预处理组。对照组未加入任何药物刺激;脂多糖组用1μg/mL脂多糖刺激细胞3 h;姜黄素预处理组分别用2.5μmol/L、10μmol/L、25μmol/L姜黄素预孵育30 min后,再加入1μg/mL脂多糖刺激细胞3 h。②原代培养的星形胶质细胞,分组方法和处理方法同①。③原代培养的小胶质细胞,随机分成对照组、脂多糖组、姜黄素25μmol/L预处理组,方法同①。处理结束后,采用real-time PCR方法检测细胞中趋化因子CXCL1和CXCL10 mRNA的表达。结果:(1)与对照组比较,脂多糖刺激C6细胞3 h时,CXCL1(P<0.001)和CXCL10(P<0.05)表达最高,因此选择脂多糖刺激细胞时间为3 h。(2)在C6细胞中,与脂多糖组比较,姜黄素25μmol/L预处理组CXCL1和CXCL10的表达均显著降低(P<0.001)。(3)在原代培养的星形胶质细胞中,与脂多糖组比较,姜黄素10μmol/L预处理组(P<0.05)、姜黄素25μmol/L预处理组(P<0.001)CXCL1的表达均显著降低,姜黄素25μmol/L预处理组CXCL10的表达显著降低(P<0.001)。(4)在原代培养的小胶质细胞中,与脂多糖组比较,姜黄素25μmol/L预处理组CXCL1和CXCL10的表达均显著降低(P<0.001)。结论:姜黄素能抑制星形胶质细胞和小胶质细胞中脂多糖诱导的趋化因子CXCL1和CXCL10的表达,具有抗炎作用。
Objective: To investigate the inhibitory effect of curcumin (CUR) on the expression of keratinocyte-derived chemokine (CXCL1) and interferon-inducible protein 10 (CXCL10) induced by lipopolysaccharide (LPS) in astrocytes and microglia. Methods:(1) Determining the time point of LPS stimulation: The cultured astrocyte line C6 cells were randomly divided into the control group, LPS 1 h, 3 h, 6 h group. The control group received no drug stimulation;LPS 1 h, 3 h, 6 h groups were stimulated by 1 μg/mL LPS for 1 h, 3 h and 6 h, respectively. The expression of chemokines CXCL1 and CXCL10 mRNA in cells was detected by real-time PCR to determine the appropriate time point of LPS stimulation.(2) CUR treatments:①C6 cells were randomly divided into 5 groups: the control group, the LPS group and the CUR 2.5 / 10 / 25 μmol/L + LPS group. The control group received no drug stimulation;the LPS group was stimulated by 1 μg/mL the LPS for 3 h;the CUR 2.5 / 10 / 25 μmol/L + LPS group were pre-incubated with 2.5 μmol/L, 10 μmol/L and 25μmol/L CUR for 30 min, and then stimulated by LPS for 3 h.②Primary cultured astrocytes, the grouping and treatments were the same as those in ①.③Primary cultured microglia were randomly divided into 3 groups: The control group, the LPS group and the CUR 25 μmol/L + LPS group. The treatments were the same as in ①. After all the treatments, the expression of chemokines CXCL1 and CXCL10 mRNA in cells was detected by real-time PCR. Results:(1) Compared with that in the control group, the expression of CXCL1 (P<0.001) and CXCL10 (P<0.05) mRNA was higher and there was a significant difference at LPS 3 h. Then the time point of LPS stimulation was selected as 3 h.(2) In C6 cells, the expression of CXCL1 and CXCL10 was decreased significantly in the 25 μM CUR + LPS group compared with the LPS group (P<0.001).(3) In the primary cultured astrocytes, the expression of CXCL1 in the 10 μM CUR + LPS group (P < 0.05) and the 25 μmol/L CUR + LPS group (P<0.001) was decreased significantly compare
作者
赵林霞
曹德利
高永静
ZHAO Linxia;CAO Deli;GAO Yongjing(Institute of Pain Medicine,Institute of Special Environmental Medicine,Nantong University,Jiangsu 226019)
出处
《交通医学》
2019年第1期1-5,共5页
Medical Journal of Communications
基金
国家自然科学基金项目(31371121
31700899)
