黄芩含药血清对脂多糖刺激大鼠原代小胶质细胞的保护作用

Inhibitory effect of serum containing Scutellaria baicalensis Georgi on activity of rat primary microglia cells stimulated by lipopolysaccharide

目的探讨黄芩提取物(SBE)含药血清对细菌脂多糖(LPS)引起的大鼠原代小胶质细胞活化的抑制作用。方法大鼠ig给予SBE 25 g·kg-1,给药后0.5～6 h眼底静脉丛取血,制备SBE含药血清。将SBE含药血清(终浓度10%)和LPS(终浓度1 mg·L-1)与大鼠原代小胶质细胞共培养24 h。采用MTT法测定细胞存活率;Griess还原法测定细胞培养液中一氧化氮(NO)含量;用超氧化物歧化酶(SOD)活性测定试剂盒测定SOD活性;分别用微量丙二醛(MDA)和谷胱甘肽(GSH)含量测定试剂盒测定MDA和GSH含量。结果在不影响细胞存活率的情况下,不同时间点的SEB含药血清可以显著降低LPS刺激的大鼠原代小胶质细胞培养液中NO和MDA的含量,SEB 0.5,1和2 h的含药血清使NO由LPS对照组的(14.2±0.8)μmol·L-1分别降低至12.9±0.6,9.2±0.6和(9.9±0.4)μmol·L-1(P<0.05);SEB 1,2和3 h的含药血清使MDA由LPS对照组的(13.4±0.7)μmol·L-1分别降低至9.42±0.64,9.13±0.57和(11.78±0.71)μmol·L-1(P<0.05);SBE 1和2 h含药血清可以显著增强培养液中SOD活性,分别由LPS对照组的(2.53±0.13)kU·L-1升高至3.52±0.18和(3.74±0.19)kU·L-1(P<0.05);SBE 1和2 h含药血清可以显著升高培养液中GSH含量,分别由LPS对照组的(7.1±1.1)mg·L-1升高至8.6±1.6和(9.2±1.7)mg·L-1(P<0.05)。结论SBE含药血清对LPS刺激的大鼠原代小胶质细胞活化具有一定的抑制作用。

OBJECTIVE To investigate the protective effect of serum containing extract of Scutellaria baicalensis Georgi （SBE） on the lipopolysaccharide-stimulated activity of primary microglia cells of rats. METHODS The rats were ig given SBE 25 g·kg^-1 and blood was taken from retinal venous plexuses within 0.5 - 6 h and the serum containing SBE was prepared. Then the primary microglia cells from normal rats were incubated with serum containing SBE （ 10% ） and LPS （ final concentration 1 mg·L ^-1 ） for 24 h. The content of nitric oxide （ NO）, malondialdehyde （ MDA）, superoxide dismutase（SOD） and glutathione（GSH） in the supernatant was examined with corresponding kits. RESULTS Serum containing SBE markedly decreased the content of NO and MDA in the LPS-stimulated primary rat microglia cells without affecting cell survival. NO content of the LPS model group was （ 14.2 ±0.8） μmol·L^-1 ; serum containing SBE within 0.5, 1 and 2 h reduced NO content to 12.9±0.6,9.2 ±0.6 and （9.9 ±0.4） μmol·L^-1 （P 〈0.05）. MDA content of the LPS model group was （ 13.4 ±0.7） μmol·L^-1 ; serum containing SBE within 1, 2 and 3 h decreased MDA content to 9.4 ± 0.6,9.1 ±0.6 and （ 11.8 ±0.7 ） μmol· L^-1 （ P 〈 0.05 ）. Also, serum containing SBE remarkably increased the content of SOD and GSH in the LPS-stimulated primary rat mi- croglia cells without affecting cell survival. SOD activity of the LPS model group was （2.53 ± 0.13）kU·L^-1 ; serum containing SBE within 1 and 2 h increased SOD activity to 3.52 ±0.18 and （3. 74 ±0. 19） kU·L^-1 （P 〈0. 05）. GSH content of the LPS model group was （7. 1 ±1.1 ）mg·L-1; serum containing SBE within 1 and 2 h increased GSH content to 8.6 ±1.6 and （9.2 ±1.7 ） g·L^-1. CONCLUSION Serum containing SBE can inhibit the activity of rat primary microglia cells stimulated by lipopolysaccharide.