Antioxidant and antifatigue effect of a standardized fraction (HemoHIM) from Angelica gigas, Cnidium officinale, and Paeonia lactiflora

Figures & data

Table 1. Primer sequences used for reverse transcriptase polymerase chain reaction (RT-PCR).

Gene	Direction	Sequence (5′ to 3′)
Nrf-2	Forward	TTCCTCTGCTGCCATTAGTCAGTC
	Reverse	GCTCTTCCATTTCCGAGTCACTG
HO-1	Forward	CTGGAAGAGGAGATAGAGCGAA
	Reverse	TCTTAGCCTCTTCTGTCACCCT
Keap1	Forward	TGCCCCTGTGGTCAAAGTG
	Reverse	AGTCCTTGGAGTCTAGCCGAG
NOQ1	Forward	GGCAGAAGAGCACTGATCGTA
	Reverse	TGATGGGATTGAAGTTCATGGC
SOD	Forward	AATGTGTCCATTGAAGATCGTGTGA
	Reverse	GCTTCCAGCATTTCCAGTCTTTGTA
CAT	Forward	TGTCCCCCACCATTGAACT
	Reverse	TCTGCAGATACCTGTGAACTG
Txn	Forward	TTCCCTCTGTGACAAGTATTCCAA
	Reverse	GGTCGGCATGCATTTGACT
GPx	Forward	CAGTTCGGACATCAGGAGAAT
	Reverse	AGAGCGGGTGAGCCTTCT
GSr	Forward	GGGCAAAGAAGATTCCAGGTT
	Reverse	GGACGGCTTCATCTTCAGTGA
GCLC	Forward	GTCCTCAGGTGACATTCCAAGC
	Reverse	TGTTCTTCAGGGGCTCCAGTC
β-actin	Forward	GCCATGTACGTAGCCATCCA
	Reverse	GAACCGCTCATTGCCGATAG

Figure 1. Chromatogram of HemoHIM using HPLC alaysis. Peak number indicated: 1: chlorogenic acid, 2: paeoniflorin, 3: nodakenin.

Figure 2. Effect of HemoHIM on levels of (A) SOD, CAT, GPx, GSr, Gclc and (B) Nrf-2, HO-1, NQO-1, Keap1 in citrinin-induced oxidative stress in L6 skeletal muscle cell. Significant difference from control (#p < 0.05, ##p < 0.01, ###p < 0.001) and from citrinin group (*p < 0.05, **p < 0.01, ***p < 0.001).

Figrue 3. Effect of HemoHIM on (A) latency time on rotarod performance and (B) swimming time to exhaustion in FST. Data are expressed as mean ± SEM. Comparison was made between control and HemoHIM groups. Significant difference from control group (*p < 0.05, **p < 0.01).

Table 2. Effect of HemoHIM on body weight after exercise performance.

Groups	Days
	0	7	14	21
Control	32.37 ± 0.88	34.02 ± 2.01	36.06 ± 2.46	37.70 ± 2.11
HemoHIM 250 mg kg^−1	32.49 ± 0.88	33.65 ± 1.33	35.46 ± 1.07	36.88 ± 0.93
HemoHIM 500 mg kg^−1	32.36 ± 1.30	33.69 ± 2.25	35.60 ± 2.40	36.85 ± 2.13
Creatine 300 mg kg^−1	31.57 ± 1.26	33.85 ± 1.41	35.30 ± 1.40	36.43 ± 1.63

Figure 4. Effect of HemoHIM on (A) glucose concentration and (B) LDH activity in serum after exercise challenge. Data are expressed as mean ± SEM. Comparison was made between control and HemoHIM groups. Significant difference from control group (*p < 0.05, **p < 0.01, ***p < 0.001).

Table 3. Effect of HemoHIM on lactate, creatine kinase, AST and ALT in serum after exercise challenge.

Group	Level in serum
	Lactate (mM)	Creatine kinase (U mL^−1)	AST (mU mL^−1)	ALT (mU mL^−1)
Control	24.18 ± 0.92	3.40 ± 0.35	75.84 ± 15.36	4.08 ± 0.73
HemoHIM 250 mg kg^−1	20.93 ± 1.70	2.99 ± 0.18	74.29 ± 14.10	3.19 ± 0.45
HemoHIM 500 mg kg^−1	21.03 ± 1.75	2.79 ± 0.37	61.50 ± 6.35	2.89 ± 0.67
Creatine 300 mg kg^−1	19.20 ± 1.09	2.71 ± 0.10	64.51 ± 4.53	3.16 ± 0.75

Data are expressed as mean ± SEM.

Figure 5. Effect of HemoHIM on (A) MDA content and (B) GPx activity in liver after exercise challenge. Data are expressed as mean ± SEM. Comparison was made between control and HemoHIM groups. Significant difference from control group (*p < 0.05, **p < 0.01, ***p < 0.001).

Figure 6. Effect of HemoHIM on (A) glycogen content and (B) LDH activity in muscle after exercise challenge. Data are expressed as mean ± SEM. Comparison was made between control and HemoHIM groups. Significant difference from control group (**p < 0.01, ***p < 0.001).

Figure 7. Effects of HemoHIM on (A) Nrf-2 and, HO-1 in liver, and (B) Nrf-2 and, HO-1 mRNA expression levels in muscle using reverse transcriptase PCR after exercise challenge. Data are expressed as mean ± SEM. Comparison was made between control and HemoHIM groups. Significant difference from control group (*p < 0.05, **p < 0.01, ***p < 0.001).