Impaired inflammatory responses in murine Lrrk2-knockdown brain microglia.
Impaired inflammatory responses in murine Lrrk2-knockdown brain microglia.
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DOI：
10.1371/journal.pone.0034693
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Joe EH
中科院分区：
文献类型：
--
作者：
Kim B;Yang MS;Choi D;Kim JH;Kim HS;Seol W;Choi S;Jou I;Kim EY;Joe EH
LRRK 2是一种帕金森病相关基因，除了神经元外，还在小胶质细胞中高度表达;然而，尚未评估其在小胶质细胞中的功能。使用Lrrk 2敲低（Lrrk 2-KD）小鼠小胶质细胞通过慢病毒介导的Lrrk 2特异性小抑制发夹RNA（shRNA）转移制备，我们发现Lrrk 2缺陷减弱脂多糖（LPS）诱导的诱导型一氧化氮合酶、TNF-α、IL-1β和IL-6的mRNA和/或蛋白表达。在Lrrk 2-KD细胞中，LPS诱导的p38丝裂原活化蛋白激酶磷酸化和NF-κ B反应性荧光素酶报告活性的刺激也降低。有趣的是，通过荧光素酶测定测量的NF-κB转录活性的降低似乎反映了抑制性NF-κB同二聚体p50/p50与DNA的结合增加。在LPS应答的HEK 293 T细胞中，人LRRK 2病理性激酶活性突变体G2019 S的过表达增加了基础和LPS诱导的磷酸化p38和JNK水平，而野生型和其他病理性（R1441 C和G2385 R）或人工激酶死亡（D1994 A）LRRK 2突变体增强或不改变基础和LPS诱导的p38和JNK磷酸化水平。然而，野生型LRRK 2和所有LRRK 2突变体同等地增强NF-κB转录活性。总之，这些结果表明，LRRK 2是小鼠小胶质细胞中炎症的正调节因子，LRRK 2突变可能会改变大脑的微环境，以促进神经炎症。
LRRK2, a Parkinson's disease associated gene, is highly expressed in microglia in addition to neurons; however, its function in microglia has not been evaluated. Using Lrrk2 knockdown (Lrrk2-KD) murine microglia prepared by lentiviral-mediated transfer of Lrrk2-specific small inhibitory hairpin RNA (shRNA), we found that Lrrk2 deficiency attenuated lipopolysaccharide (LPS)-induced mRNA and/or protein expression of inducible nitric oxide synthase, TNF-α, IL-1β and IL-6. LPS-induced phosphorylation of p38 mitogen-activated protein kinase and stimulation of NF-κB-responsive luciferase reporter activity was also decreased in Lrrk2-KD cells. Interestingly, the decrease in NF-κB transcriptional activity measured by luciferase assays appeared to reflect increased binding of the inhibitory NF-κB homodimer, p50/p50, to DNA. In LPS-responsive HEK293T cells, overexpression of the human LRRK2 pathologic, kinase-active mutant G2019S increased basal and LPS-induced levels of phosphorylated p38 and JNK, whereas wild-type and other pathologic (R1441C and G2385R) or artificial kinase-dead (D1994A) LRRK2 mutants either enhanced or did not change basal and LPS-induced p38 and JNK phosphorylation levels. However, wild-type LRRK2 and all LRRK2 mutant variants equally enhanced NF-κB transcriptional activity. Taken together, these results suggest that LRRK2 is a positive regulator of inflammation in murine microglia, and LRRK2 mutations may alter the microenvironment of the brain to favor neuroinflammation.
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影响因子：
8.6
作者：
Covy, Jason P.;Yuan, Wuxing;Waxman, Elisa A.;Hurtig, Howard I.;Van Deerlin, Vivianna M.;Giasson, Benoit I.
通讯作者：
Giasson, Benoit I.
影响因子：
25
作者：
Li, Yanping;Liu, Wencheng;Oo, Tinmarla F.;Wang, Lei;Tang, Yi;Jackson-Lewis, Vernice;Zhou, Chun;Geghman, Kindiya;Bogdanov, Mikhail;Przedborski, Serge;Beal, M. Flint;Burke, Robert E.;Li, Chenjian
通讯作者：
Li, Chenjian
影响因子：
4.7
作者：
He, Y;Imam, SZ;Le, WD
通讯作者：
Le, WD
影响因子：
3.7
作者：
Heo, Hye Young;Park, Ji-Min;Seol, Wongi
通讯作者：
Seol, Wongi
影响因子：
9.3
作者：
Bachstetter, Adam D.;Xing, Bin;Van Eldik, Linda J.
通讯作者：
Van Eldik, Linda J.