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Table 2 Effect of signal transduction inhibitors on ET-induced expression of CCL2, CXCL1 and CX3CL1 mRNA

From: Different actions of endothelin-1 on chemokine production in rat cultured astrocytes: reduction of CX3CL1/fractalkine and an increase in CCL2/MCP-1 and CXCL1/CINC-1

 

Ratio of chemokine to G3PDH mRNA copy number

 

(% of no treatment)

 

CCL2/MCP-1

CXCL1/CINC-1

CX3CL1/fractalkine

no treatment

100.0 ± 31.4 (20)

100.0 ± 22.0 (20)

100.0 ± 17.5 (24)

100 nM ET-1

313.8 ± 65.1 (20)a

404.7 ± 63.9 (20)a

37.5 ± 8.5 (23)a

+ 30 μM BAPTA/0.5 mM EGTA

307.4 ± 89.0 (7)

342.7 ± 65.0 (7)

122.8 ± 25.2 (9)b

+ 10 nM staurosporine

444.8 ± 122.3 (6)

385.4 ± 79.0 (6)

108.6 ± 25.6 (4)b

+ 100 μM PDTC

129.2 ± 16.5 (12)b

140.2 ± 20.7 (12)b

42.5 ± 8.8 (18)

+ 10 μM SN50

188.7 ± 64.3(8)b

136.1 ± 22.6 (8)b

54.6 ± 23.9 (11)

+ 500 nM mithramycin

37.8 ± 9.2 (8)c

49.8 ± 24.1 (8)c

36.3 ± 11.7 (10)

+ 50 μM PD98059

277.4 ± 90.7 (12)

360.2 ± 103.2 (12)

49.6 ± 7.2 (10)

+ 20 μM SB203580

118.3 ± 36.9 (13)c

229.1 ± 73.6 (13)b

37.1 ± 6.3 (20)

+ 1 μM SP600125

110.2 ± 23.6 (8)b

188.5 ± 22.5 (8)b

47.1 ± 22.0 (10)

  1. a P <0.01 versus no treatment, b P <0.05, c P <0.01, versus 100 nM ET-1 by one-way ANOVA followed by Fisher’s PLSD test. Astrocytes were treated with 100 nM ET-1 in the presence of the signal transduction inhibitors indicated. Total RNA was extracted at one hour (for CCL2 and CXCL1) or six hours (for CX3CL1) after the addition of ET-1. The inhibitors were included in the serum-free medium for 30 minutes before the addition of ET-1. The copy numbers of CCL2, CXCL1 and CX3CL1 mRNA was normalized to G3PDH. Results are the mean ± SEM and the numbers of experiments are indicated in parentheses. ANOVA, analysis of variance; BAPTA/AM, 1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; ET, endothelin; G3PDH, glyceraldehyde-3-phosphate dehydrogenase; PDCT, pyrrolidine dithiocarbamate; PLSD, protected least significant difference; SEM, standard error of the mean.