Influences of Hyriopsis cumingii polysaccharides on mice immunosignaling molecules and T lymphocyte differentiation

Figures & data

Table 1. Prime sequences of IL-2, IL-10, IFN-γ, CD4 and CD8 used in RTFQ-PCR.

Group	Prime sequences
IL-2	Forward: 5′- CGG CAT GTT CTG GAT TTG AC -3′
	Reverse: 5′- CAT CAT CGA ATT GGC ACT CA -3′
IL-10	Forward: 5′- GGA AGA GAA ACC AGG GAG AT -3′
	Reverse: 5′- GCA GAC AAA CAA TAC ACC ATT C -3′
IFN-γ	Forward: 5′- CTG CTG ATG GGA GGA GAT GT -3′
	Reverse: 5′- ATT TGT CAT TCG GGT GTA GTC A -3′
CD4	Forward: 5′- GCA CAG CTA TCA CGG CCT AT -3′
	Reverse: 5′- ATC CTT CTC TGC CTT CCA CA -3′
CD8	Forward: 5′- GCC ATG AGG GAC ACG AAT AA -3′
	Reverse: 5′- GAC TGG CAC GAC AGA ACT GA -3′

Figure 1. Electrophoresis spectra of DNA marker and RTFQ-PCR products of endogenous reference gene (β-actin) and target genes (IL-2, CD4, CD8, IL-10 and IFN-γ). Channel M: DNA marker; Channel 1: IL-2 (about 120 bp); Channel 2: CD4 (about 115 bp); Channel 3: CD8 (about 130 bp); Channel 4: β-actin (about 275 bp and 175 bp); Channel 5: IL-10 (about 200 bp); Channel 6: IFN-γ (about 180 bp).

Table 2. Effects of crude HCPS on the content of IL-2, IL-10, IFN-γ, iNOS, CD4, CD8 and cAMP in spleen in vivo by ELISA.

Item	Relative level
	II	III	IV	V
IL-2	0.6891 ± 0.05596^a	0.7148 ± 0.04174^ab	0.8409 ± 0.03919^b	1.1392 ± 0.12088^c
IL-10	1.4842 ± 0.12994^a	1.2612 ± 0.10921^b	0.8598 ± 0.05807^c	0.7010 ± 0.08737^c
IFN-γ	0.4486 ± 0.07006^a	0.5567 ± 0.07972^a	0.7513 ± 0.09199^b	0.9237 ± 0.10338^c
iNOS	0.6731 ± 0.06189^a	0.8601 ± 0.07585^ab	0.9746 ± 0.06807^b	1.2371 ± 0.21320^c
cAMP	2.6773 ± 0.19415^a	2.1319 ± 0.12596^b	1.9405 ± 0.10230^b	1.6520 ± 0.12110^c
CD4	0.7895 ± 0.05325^a	0.8949 ± 0.09288^a	1.1565 ± 0.02110^b	1.3460 ± 0.07704^c
CD8	0.8228 ± 0.03087^a	0.8860 ± 0.09264^a	1.0359 ± 0.04939^b	1.1326 ± 0.08654^b

Notes: Group I: control group; Group II: model group; Group III: low dose of crude HCPS (200 mg/kg BW); Group IV: medium dose of crude HCPS (400 mg/kg BW); Group V: high dose of crude HCPS (800 mg/kg BW). Different superscript (a, b, c) indicted there was significant difference (P < .05) between groups. Same superscript suggested difference between groups was not significant (P > .05).

Table 3. Effects of crude HCPS on the mRNA synthesis of IL-2, IL-10, IFN-γ, CD4 and CD8 in spleen in vivo by RTFQ-PCR.

Item	Relative mRNA content
	II	III	IV	V
IL-2	0.7247 ± 0.08306^a	0.8151 ± 0.10721^a	1.1291 ± 0.10331^b	1.3377 ± 0.06460^c
IL-10	1.4478 ± 0.11038^a	1.2368 ± 0.10144^b	0.8120 ± 0.06084^c	0.7001 ± 0.07228^c
IFN-γ	0.4179 ± 0.07605^a	0.5298 ± 0.07325^a	0.7177 ± 0.08172^b	0.8928 ± 0.11420^c
CD4	0.7604 ± 0.09076^a	0.9431 ± 0.05367^b	1.2445 ± 0.08353^c	1.4450 ± 0.06135^d
CD8	0.8554 ± 0.04994^a	0.9105 ± 0.09800^a	1.1361 ± 0.11767^b	1.2508 ± 0.11022^b

Notes: Group I: control group; Group II: model group; Group III: low dose of crude HCPS (200 mg/kg BW); Group IV: medium dose of crude HCPS (400 mg/kg BW); Group V: high dose of crude HCPS (800 mg/kg BW). Different superscript (a, b, c, d) indicted there was significant difference (P < .05) between groups. Same superscript suggested difference between groups was not significant (P > .05).

Table 4. Effects of crude HCPS on the content of IL-10 and cGMP in serum in vivo by ELISA.

Item	Relative level
	II	III	IV	V
IL-10	1.1899 ± 0.11562^a	0.9509 ± 0.05917^b	0.4841 ± 0.06754^c	0.4010 ± 0.06015^c
cGMP	0.8881 ± 0.08313^a	1.1410 ± 0.05514^b	1.4954 ± 0.14733^c	1.7650 ± 0.07249^d

Notes: Group I: control group; Group II: model group; Group III: low dose of crude HCPS (200 mg/kg BW); Group IV: medium dose of crude HCPS (400 mg/kg BW); Group V: high dose of crude HCPS (800 mg/kg BW). Different superscript (a, b, c, d) indicted there was significant difference (P < .05) between groups. Same superscript suggested difference between groups was not significant (P > .05).

Figure 2. Effects of HCPS on the gene expression of iNOS in peritoneal macrophage in vitro by ELISA. Group I: control group; Group II: low dose of HCPS (50 μg/mL); Group III: medium dose of HCPS (100 μg/mL); Group IV: high dose of HCPS (200 μg/mL). Different letter (a, b, c, d, e, f) indicted there was significant difference (P < .05) between groups. Same letter suggested difference between groups was not significant (P > .05).