The effects of highly bioavailable curcumin in downhill running-induced muscle damage model

Figures & data

Table 1. Change of body weight of mice.

		DHR
Group	Resting (n = 8)	Saline (n = 8)	TS-P1^16.7 (n = 8)	TS-P1^50 (n = 8)	CR-033P^16.7 (n = 8)	CR-033P^50 (n = 8)	curcumin^50 (n = 8)
Before DHR	22.025 ± 0.21	22.075 ± 0.38	22.175 ± 0.34	21.938 ± 0.23	21.944 ± 0.16	22.063 ± 0.32	21.988 ± 0.34
After resting	*22.725 ± 0.20
After DHR		22.050 ± 0.34	†21.725 ± 0.30	21.913 ± 0.29	21.688 ± 0.22	†21.663 ± 0.27	22.400 ± 0.28

Measurement of average body weight (g) of mice before DHR, and after resting or DHR. Data were expressed as mean ± SEM (n = 8, *p < .05 for “before DHR” vs “after resting”, and ^†p < .05 for “before DHR” vs “after DHR” by paired t-test).

Table 2. DNA primers for real-time RT-PCR.

Targets	Forward primer (5’-3’)	Reverse primer (5’-3’)
glyceraldehyde-3-phosphate dehydrogenase (Gapdh)	TGAAGCAGGCATCTGAGGG	CGAAGGTGGAAGAGTGGGAG
NADPH oxidase 2 (Nox2)	TTGGGTCAGCACTGGCTCTG	TGGCGGTGTGCAGTGCTATC
Interleukin 1β (Il1b)	TGGACCTTCCAGGATGAGGACA	GTTCATCTCGGAGCCTGTAGTG
Interleukin 6 (Il6)	TACCACTTCACAAGTCGGAGGC	CTGCAAGTGCATCATCGTTGTTC
Interleukin 10 (Il10)	CGGGAAGACAATAACTGCACCC	CGGTTAGCAGTATGTTGTCCAGC
tumor necrosis factor-α (Tnfa)	GGTGCCTATGTCTCAGCCTCTT	GCCATAGAACTGATGAGAGGGAG
prostaglandin-endoperoxide synthase 2 (Ptgs2)	AGAAGGAAATGGCTGCAGAA	GCTCGGCTTCCAGTATTGAG
monocyte chemoattractant protein 1 (Mcp1)	CTTCTGGGCCTGCTGTTCA	CCAGCCTACTCATTGGGATCA
C-X-C motif chemokine ligand 14 (Cxcl14)	CCAAGATTCGCTATAGCGAC	CCTGCGCTTCTCGTTCCAGG
adhesion G protein-coupled receptor E1 (Adgre1)	CTTTGGCTATGGGCTTCCAGTC	GCAAGGAGGACAGAGTTTATCGTG

Sequences of each DNA oligomer producing 100–200 bp PCR products from cDNA reverse-transcribed from mRNAs in gastrocnemius muscle.

Figure 1. Antioxidant effect of TS-P1 and CR-033P in vitro (a, b) measurement of scavenging activity of TS-P1 (a) and CR-033P (b) against DPPH radical. Ascorbic acid (AA) is used as a positive control. (c, d) measurement of ferric reducing antioxidant power (FRAP) of TS-P1 (c) and CR-033P (d). (e, f) measurement of total antioxidant capacity (TAC) of TS-P1 (e) and CR-033P (f). Data were expressed as mean ± SEM (n = 3, *p < .05, **p < .01, ***p < .001 vs untreated group by one-way ANOVA using Dunnett’s multiple comparison test for TS-P1 and CR-033P; n = 3, ***p < .001 vs untreated group by t-test for AA).

Figure 2. Muscle-protective effect of TS-P1 and CR-033P in mouse DHR model. (a) Experimental scheme of mouse DHR model. For two days, mice were trained to run on flat treadmill for 10 min per day, and then subjected to DHR after 1 day resting. (b, c) measurement of activities of lactate dehydrogenase (LDH) (b) and creatine kinase (CK) (c) in plasma of mice treated with or without saline, TS-P1^16.7, TS-P1⁵⁰, CR-033P^16.7, CR-033P⁵⁰, and curcumin⁵⁰. Data were expressed as mean ± SEM (n = 8, *p < .05 for resting and ^†p < .05 for saline by t-test).

Figure 3. Antioxidant effect of TS-P1 and CR-033P upon DHR-induced oxidative stress (a, b) measurement of levels of hydrogen peroxide (H₂O₂) (a) and malondialdehyde (MDA) (b) in gastrocnemius muscle (GA) of mice treated with or without saline, TS-P1^16.7, TS-P1⁵⁰, CR-033P^16.7, CR-033P⁵⁰, and curcumin⁵⁰. Data were expressed as mean ± SEM (n = 8, *p < .05 for resting by t-test). (c) Relative level of mRNA of Nox2 in GA of mice treated with or without saline, TS-P1^16.7, TS-P1⁵⁰, CR-033P^16.7, CR-033P⁵⁰, and curcumin⁵⁰. Data were expressed as mean ± SEM (n = 8, *p < .05 for resting and †p < .05 for saline by t-test). (d, e) measurement of activities of glutathione peroxidase (GPx) (D) and catalase (E) in GA of mice treated with or without saline, TS-P1^16.7, TS-P1⁵⁰, CR-033P^16.7, CR-033P⁵⁰, and curcumin⁵⁰. Data were expressed as mean ± SEM (n = 8, *p < .05 for resting and ^†p < .05 for saline by t-test).

Figure 4. Anti-inflammatory effect of TS-P1 and CR-033P upon DHR. (a-c) measurement of relative levels of mRnas of Il1b (a), Il6 (b), and tnfa (c), coding pro-inflammatory cytokines IL-1β, IL-6 and TNFα respectively, in GA of mice treated with or without saline, TS-P1^16.7, TS-P1⁵⁰, CR-033P^16.7, CR-033P⁵⁰, and curcumin⁵⁰. Data were expressed as mean ± SEM (n = 8, *p < .05 for resting and †p < .05 for saline by Mann – Whitney rank sum test). (d) Measurement of relative level of mRNA of Il10, coding IL-10, in GA of mice treated with or without saline, TS-P1^16.7, TS-P1⁵⁰, CR-033P^16.7, CR-033P⁵⁰, and curcumin⁵⁰. Data were expressed as mean ± SEM (n = 8, *p < .05 for resting and †p < .05 for saline by Mann – Whitney rank sum test). (e) Measurement of relative levels of mRnas of Ptgs2, coding COX-2, in GA of mice treated with or without saline, TS-P1^16.7, TS-P1⁵⁰, CR-033P^16.7, CR-033P⁵⁰, and curcumin⁵⁰. Data were expressed as mean ± SEM (n = 8, *p < .05 for resting and ^†p < .05 for saline by Mann – Whitney rank sum test).

Figure 5. Inhibitory effect of TS-P1 and CR-033P on macrophage activation upon DHR. (a-c) measurement of relative levels of mRnas of Mcp1 (a), Cxcl14 (b), and Adgre1 (c), which are genes involved in macrophagic activation, in GA of mice treated with or without saline, TS-P1^16.7, TS-P1⁵⁰, CR-033P^16.7, CR-033P⁵⁰, and curcumin⁵⁰. Data were expressed as mean ± SEM (n = 8, *p < .05 for resting and †p < .05 for saline by Mann – Whitney rank sum test).

Data availability statement

The raw datasets used and/or analyzed are available from the corresponding author upon reasonable request.