**9. Effects of HE-86 administration on localization of NF–κB in renal tissue**

Immunohistochemical analysis was performed to determine the localization of NF–κB in the renal cortex (Fig.1-2). NF-B, a critical transcriptional factor for controlling inflammatory response, has been shown to play a central role in inflammatory diseases, including kidney diseases [33]. In normal rats, only tubular epithelial cells were weakly stained by the monoclonal anti-NF–κB antibody, while glomeruli were hardly stained. In control uraemic rats, however, proximal tubular epithelial cells, especially of dilated tubules, were intensively stained by the anti-NF–κB antibody. In contrast, in the HE-86-treated uraemic rats activation of the NF-B in tubular epithelial cells was less prominent as compared with that in the control uraemic rats. The staining of NF–κB as shown in the control uraemic rats found increased NF–κB -positive (intensively stained) area in the renal cortex, whereas HE-86-treated rats showed markedly decreased NF–κB -positive area as compared to the control uraemic rats. These data demonstrate that HE-86 markedly reduces the overexpress of NF– κB on the remnant tubular cells.

Molecular Mechanisms of Nephro-Protective

**0**

TNF–

of glomerular and tubulointerstitial NF–

glomerular and tubulointerstitial NF–

compared to the normal sham-control.

**R in renal tissue** 

**2**

**4**

**6**

**8**

**10**

**12**

Action of HE-86 Liquid Extract in Experimental Chronic Renal Failure 183

Sham group Control group Treatment group

\*

A B C

the glomerulus and tubulointerstitium using the Quantitative Image System. A: Percentage

B localization in glomerular and tubulointerstitial without treatment C: Percentage of

was decreased significantly. Each bar represents data (mean ± SEM) #, P < 0.05 and ##, P < 0.001, when compared to empty vector-treated controls; \*, P < 0.05 and \*\*, P < 0.01, when

**10. Effects of HE-86 administration on mRNA levels of TNF–α, Ang II and AT II** 

The effects of HE-86 on the gene expression of Ang II (Figure 3), AT1R (Figure 4) and

(Figure 5) in the renal cortex were examined. We investigated the potential

Fig. 2. Semiquantitative analysis of the therapeutic effect of HE-86 on NF–

##

B deposition in sham group. B: Percentage of NF–

B accumulation in twelve rats treated with HE-86

B localization in

Fig. 1. Immunohistochemistry demonstrates that HE-86 inhibits renal NF-B accumulation within the kidney. The accumulation of NF-B in the glomerular and tubulointerstitium is markedly increased in empty vector-treated animals (C, D), compared to normal shamcontrols (A,B), which is substantially inhibited in 5/6 nephrectomized rats treated with HE-86 (E, F). Original magnifications, x100.

A B

C D

E F

markedly increased in empty vector-treated animals (C, D), compared to normal shamcontrols (A,B), which is substantially inhibited in 5/6 nephrectomized rats treated with HE-

B in the glomerular and tubulointerstitium is

B accumulation

Fig. 1. Immunohistochemistry demonstrates that HE-86 inhibits renal NF-

within the kidney. The accumulation of NF-

86 (E, F). Original magnifications, x100.

Fig. 2. Semiquantitative analysis of the therapeutic effect of HE-86 on NF–B localization in the glomerulus and tubulointerstitium using the Quantitative Image System. A: Percentage of glomerular and tubulointerstitial NF–B deposition in sham group. B: Percentage of NF– B localization in glomerular and tubulointerstitial without treatment C: Percentage of glomerular and tubulointerstitial NF–B accumulation in twelve rats treated with HE-86 was decreased significantly. Each bar represents data (mean ± SEM) #, P < 0.05 and ##, P < 0.001, when compared to empty vector-treated controls; \*, P < 0.05 and \*\*, P < 0.01, when compared to the normal sham-control.
