Antioxidant and Antifibrotic Effect of a Herbal Formulation In Vitro

The Korean herbal formulation Ojayeonjonghwan is used for improving late-onset hypogonadism (LOH) symptoms such as erectile dysfunction (ED). A previous research suggested that a modified Ojayeonjonghwan (KH-204) could be used as an alternative to the treatment for ED. The pharmacological effects were examined in different conditions, including in vitro and in vivo. We measured the survival rate of TM3 Leydig cells under the oxidative stress condition. The s.c. injection of leuprorelin was used to induce androgen deprivation. We measured serum testosterone levels, oxidative stress, and apoptosis. The results of the treatment by KH-204 (1) preserved TM3 cells from oxidative stress by improving the expression of nuclear factor erythroid 2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1); (2) lowered the expression of transforming growth factor-beta (TGF-β) 1/SMAD; (3) increased the average of serum testosterone in androgen-deprived male rats; (4) kept the activation of spermatogenesis; (5) upgraded the contents of 8-hydroxy-20-deoxyguanosine (8-OHdG) and degraded the contents of superoxide dismutase (SOD); and (6) reduced apoptosis. We studied that KH-204 improved testicular dysfunction in LOH. It is likely, at least in part, to degrade oxidative stress through the Nrf2/HO-1 pathway. These findings may offer credible evidences for the use of new alternative therapies to treat LOH

Modulation of MMP expression in mice by the three garlic compounds.

<p>(A) Western blot analysis was performed to detect MMP3, MMP9, MMP12, and TIMP4 protein levels in cytosolic extract of the skins of from hairless mice. (B) Blots were quantified by densitometry as percentages of the control. One-factor ANOVA was used to determine significance: ^# p<0.05 vs. control, *p<0.05 vs. the UVB-irradiated group; CA, caffeic acid; MMP, matrix metalloproteinase; SAC, <i>S</i>-allyl cysteine; TIMP4, tissue inhibitor of metalloprotease4; UVB, ultraviolet B.</p

Structures of active the three garlic compounds.

<p>(A) caffeic acid, (B) <i>S</i>-allyl cysteine, (C) uracil.</p

Possible mechanism of active three garlic compounds about effects on anti-wrinkle.

<p>COX-2, cyclooxygenase-2; iNOS, inducible nitric oxide synthase; MMP1, matrix metalloproteinase1; MMP3, metalloproteinase3; MMP9, metalloproteinase9; MMP13, matrix metalloproteinase13; UVB, ultra violet B.</p

Effects of the three garlic compounds on type І procollagen level by UVB in hairless mice.

<p>(A) Western blotting was performed to detect cytoplasmic extracts of UVB-irradiated hairless mouse skin. (B) Blots were quantified by densitometry as percentages of control. One-factor ANOVA were used to determine significance: ^*p<0.05 vs. control. CA, caffeic acid; SAC, <i>S</i>-allyl cysteine, UVB, ultraviolet B.</p

Effects of the three compounds on intracellular ROS/ONOO¯ in UVB-irradiated hairless mice.

<p>(A) (B) ROS and ONOO<b>¯</b> generations by UVB in hairless mouse skins were decreased concentration-dependently by pretreating animals with each of the three compounds. Results are expressed as means±SEs of three determinations. Significance was determined using one-factor ANOVA: ^{# # #} p < 0.001, ^{# #} p < 0.01 vs. control; *p < 0.05, **p < 0.01 and ***p < 0.001 vs. UVB-irradiated group; CA, caffeic acid; COX-2, cyclooxygenase-2; iNOS, inducible nitric oxide synthase; MMP1, matrix metalloproteinase1; MMP3, metalloproteinase3; MMP9, metalloproteinase9; MMP13, matrix metalloproteinase13; ONOO¯, peroxynitrite; ROS, reactive oxygen species ; UVB, ultraviolet B.</p

Changes in the activities of nuclear NF-κB and AP-1 caused by three compounds in UVB-irradiated hairless mice.

<p>(A) Western blotting was performed to determine nuclear p50, p-p65, Ac-p65, p65, cFOS, and p-cJun protein levels in nuclear extracts of the skins of UVB-irradiated hairless mice. The protein levels of NF-κB family members and of AP-1 component increased after UVB irradiation, but these increases were reduced by all three compounds. (B) Blots were quantified by densitometry as percentages of the control. One-factor ANOVA was used to determine significance: ^{# # #} p < 0.001 vs. control, ^*p<0.05, **p < 0.01 and ***p < 0.001 vs. UVB-irradiated group respectively. AP-1, activator protein 1; UVB, ultraviolet B.</p

Active compounds from garlic decreased wrinkle depth in hairless mice.

<p>(A) Representative images of skin replicas showing that the histological formation of wrinkles was reduced by the three garlic compounds as compared with UVB-treated animals, (B) wrinkle area (%), (C) wrinkle length, and (D) wrinkle depth.</p

Inhibition of the expressions of NF-κB-dependant genes by the three garlic compounds.

<p>(A) To determine the effect of UVB on the expressions of NF-κB dependent genes, we examined the expressions of COX-2 and iNOS <i>in </i><i>vivo</i>. (B) Blots were quantified by densitometry as percentages of the control. One-factor ANOVA was used to determine significance: ^{# # #} p < 0.001 vs. control, ^*p<0.05, **p < 0.01 and ***p < 0.001 vs. UVB-irradiated group.</p