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Journal of Chinese Integrative Medicine ›› 2011, Vol. 9 ›› Issue (2): 201-208.doi: 10.3736/jcim20110214

• Original Experimental Research • Previous Articles     Next Articles

Effects of vitamin E on mercuric chloride-induced renal interstitial fibrosis in rats and the antioxidative mechanism

Yan-yan Tao1, Qing-lan Wang1, Ji-li Yuan1, Li Shen1, Cheng-hai Liu1,2,3,4()   

  1. 1. Institute of Liver Diseases, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
    2. Key Laboratory of Liver and Kidney Diseases of Ministry of Education, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
    3. E-institute of Internal Medicine of Traditional Chinese Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
    4. Shanghai Key Laboratory of Traditional Chinese Clinical Medicine, Shanghai 201203, China
  • Received:2010-09-21 Accepted:2010-11-11 Online:2011-02-20 Published:2011-02-15
  • Contact: Liu Cheng-hai

Objective: To observe the effects of vitamin E (Vit E) on mercuric chloride (HgCl2)-induced renal interstitial fibrosis (RIF) in rats and discuss its antioxidative mechanism.
Methods: A total of 32 Sprague-Dawley rats were randomly assigned to three groups: normal group, model group and Vit E group. RIF was induced by oral administration of HgCl2 at a dose of 8 mg/kg body weight once a day for 9 weeks. Rats in Vit E group were administered with Vit E capsule at 100 mg/kg body weight, and rats in normal and model groups were treated with normal saline. At the end of the 9th week, rats were sacrificed and renal hydroxyproline (Hyp) content was assayed with Jamall’s method and collagen deposition was visualized by using hematoxylin and eosin (HE), Masson’s trichrome and periodic acid-silver methenamine (PASM) staining. The activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), and contents of glutathione (GSH) and malondialdehyde (MDA) in kidney tissue were tested with commercial kits. The expressions of nuclear factor-κB (NF-κB), inhibitor-κB (IκB), phospho-IκB (p-IκB) and tumor necrosis factor-α (TNF-α) were determined by Western blot. The expression of α-smooth muscle actin (α-SMA) was assayed by Western blot and immunofluorescent staining.
Results: Renal Hyp content, HE, Masson’s trichrome and PASM staining results and α-SMA expression confirmed development of HgCl2-induced RIF in rats. Oxidative stress markers GSH, GSH-Px and MDA confirmed oxidative stress in RIF rats. Compared with model rats, rats in Vit E group had lower kidney Hyp content (P<0.01). GSH and MDA contents decreased significantly in Vit E group compared with model group (P<0.01). The expressions of NF-κB and IκB had no significant difference among all groups (P>0.05). In Vit E group, the expressions of p-IκB and TNF-α decreased significantly compared with model group (P<0.01). The expression of α-SMA in Vit E group was also decreased significantly compared with model group (P<0.01).
Conclusion: Vit E has a protective effect on experimental RIF induced by HgCl2 in rats and it is related to inhibition of lipid peroxidation, which involves blocking of NF-κB signaling pathway and the activation of cells producing extracellular matrix.

Key words: vitamin E, renal interstitial fibrosis, mercuric chloride, lipid peroxidation, animal experimentation, rats

Figure 1

Effects of vitamin E on histological changes in rats’ kidneys tested by HE, Masson’s trichrome and PASM staining (Light microscopy, ×200) Normal: normal kidney; Model: atrophy tubules, thickened membrane, and widened interstitial space, more inflammatory cells infiltration and more myofibroblasts were observed; Vit E: less inflammatory cells infiltration and some dilated tubules were founded, with no tubules atrophy and less renal interstitial fibrosis. HE: hematoxylin and eosin; PASM: periodic acid-silver methenamine."

Figure 2

Effects of vitamin E on α-SMA expression in rats’ kidneys A: Immunofluorescence staining of α-SMA in rats’ kidneys (Fluorescence microscopy, ×200). Normal: α-SMA-positive renal fibroblasts (red, as shown by the green arrow) were weakly stained and were detected in tubuli and glomeruli. Model: massive α-SMA-positive renal fibroblasts were found in fibrotic kidneys compared with normal rats. Vit E: α-SMA-positive renal fibroblasts decreased remarkably. B: Western blot showed that α-SMA expression in normal rats was little, whereas it increased significantly in model rats. In Vit E group, the expression of α-SMA decreased significantly. C: Graphic presentation of the relative expression of α-SMA. The values are expressed as the density of α-SMA to GAPDH (%). **P<0.01, vs normal group; △△P<0.01, vs model group. α-SMA: α-smooth muscle actin; GAPDH: glyceraldehyde-3-phosphate dehydrogenase."



Figure 3

Effects of vitamin E on activation of NF-κB signal pathway The values are expressed as the density of TNF-α (p-IκB) to GAPDH (%). **P<0.01, vs normal group; △P<0.05, vs model group. NF-κB: nuclear factor-κB; IκB: inhibitor-κB; p-IκB: phospho-IκB; TNF-α: tumor necrosis factor-α; GAPDH: glyceraldehyde-3-phosphate dehydrogenase."

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