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目的 :观察脂多糖 (LPS)对体外培养的系膜细胞 (GMC)产生NO的影响及这一过程对GMC增殖的影响。方法 :用培养的第一、二代SD大鼠肾小球系膜细胞进行实验 ,分别加入LPS或LPS加白细胞介素 - 1受体拮抗剂 (IL - 1ra) ,2 4h后用经典的Griess方法测定培养上清中亚硝酸盐含量 ,用 [3 H]-TdR掺入率测定GMC的增殖 ,狭缝和Northern杂交测定iNOSmRNA表达。结果 :LPS刺激后 ,GMC出现iNOSmRNA的表达及亚硝酸盐增多 [(0 6 4± 0 2 5 )nmmol/ 10 4 细胞vs (0 12± 0 0 6 )nmmol/ 10 4 细胞 ],GMC增殖 (3735± 1177 9vs 1785± 2 80 6 ) ;LPS加IL - 1ra组 ,iNOSmRNA表达比LPS组减少 40 % ,上清亚硝酸盐含量更高 [(3 2 8± 0 33)nmmol/ 10 4 细胞 ],GMC增殖被抑制 (818± 77 2 7)。结论 :LPS诱导GMCiNOSmRNA表达及亚硝酸盐增多 ,并有GMC增殖。IL - 1ra部分拮抗iNOSmRNA的表达 ,但亚硝酸盐产量更高 ,GMC增殖被抑制
Objective: To observe the effect of lipopolysaccharide (LPS) on nitric oxide (NO) production in cultured mesangial cells (GMC) and the effect of LPS on GMC proliferation. Methods: The cultured glomerular mesangial cells of first and second generation SD rats were added into LPS or LPS plus interleukin - 1 receptor antagonist (IL - 1ra) respectively. After 24 hours, the cells were cultured with classic Griess Methods The content of nitrite in culture supernatant was measured. The proliferation of GMC was measured by [3 H] -TdR incorporation, and the expression of iNOS mRNA was detected by slit and Northern blot. RESULTS: After LPS stimulation, the expression of iNOS mRNA and the increase of nitrite in GMC [(0 64 ± 0 2 5) nmmol / 10 4 cells vs (0 12 ± 0 0 6) nmmol / 10 4 cells] 3735 ± 1177 9vs 1785 ± 2 806). Compared with LPS group, the expression of iNOS mRNA in LPS plus IL - 1ra group was decreased by 40% and the content of nitrite in supernatant was higher than that in the control group [(3 2 8 ± 0 33) nmmol / 10 4 cells] , GMC proliferation was inhibited (818 ± 77 2 7). CONCLUSION: LPS induces GMCiNOS mRNA expression and nitrite increase with GMC proliferation. IL - 1ra partially antagonized the expression of iNOS mRNA but nitrite production was higher and GMC proliferation was inhibited