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Journal of Chinese Integrative Medicine ›› 2009, Vol. 7 ›› Issue (1): 59-64.doi: 10.3736/jcim20090109

• Original Experimental Research • Previous Articles     Next Articles

Interventional effects of emodin on transforming growth factor-beta1/integrin-linked kinase signal way in interleukin-1β-induced transdifferen-tiation of rat tubular epithelial-myofibroblasts

 Ting-fang Chena, Ming Chena, Jian-hua Qina, Ming-yong Wangb   

  1. a Department of Nephrology, Affiliated Hospital, Luzhou Medical College, Luzhou, Sichuan Province 646000, China
    b Laboratory of Infection and Immunity, Affiliated Hospital, Luzhou Medical College, Luzhou,Sichuan Province 646000, China
  • Received:2008-07-10 Accepted:2008-09-29 Online:2009-01-20 Published:2009-01-15

Objective

To study the effects of transforming growth factor-beta1/integrin-linked kinase (TGF-β1/ILK) signal way in interleukin-1β (IL-1β)-induced rat tubular epithelial-myofibroblast transdifferentiation (TEMT), and to investigate whether emodin inhibits IL-1β-induced TEMT through the TGF-β1/ILK signal way-dependent mechanism.
Methods

Normal rat kidney epithelial cell line (NRK52E) was used in this study. NRK52E cells were divided into blank control group, emodin control group, IL-1β-induced group, emodin-inhibited group, SB431542 (TGF-β1 type Ⅰ receptor blocker)-inhibited group, emodin plus SB431542-inhibited group, emodin-pretreated group and emodin-reversed group. After 48-hour culture, morphological changes of the NRK52E cells were observed by an inverted phase contrast microscope. The expressions of alpha-smooth muscle actin (α-SMA) and E-cadherin were detected by two-color immunohistochemical staining, while the expressions of TGF-β1 and ILK were detected by one-color immunohistochemical staining. We also performed the imaging analysis to quantitatively analyze the result of the immunohistochemical staining. The secretion of fibronectin (FN) was analyzed by enzyme-linked immunosorbent assay.
Results

 Compared with the blank control group, IL-1β might induce TEMT, which was showed in increasing expression of α-SMA, increasing secreting of FN and decreasing expression of E-cadherin, and at the same time the expressions of TGF-β1 and ILK were enhanced (P<0.05). Emodin might inhibit all of those changes induced by IL-1β (P<0.05). When TGF-β1 signal way was intercepted, IL-1β induced-TEMT was suppressed and the expression of ILK was decreased, however, there was no significant difference in expression of TGF-β1 between the SB431542 group and the IL-1β-induced group. Compared with emodin-inhibited group, emodin-pretreatment could not prevent IL-1β induced-TEMT in a certain extent, but emodin could not revert IL-1β-induced TEMT. Spearman correlation analysis showed that TGF-β1 expression had positive correlation with expressions of α-SMA, FN, ILK and negative correlation with E-cadherin expression, and the expression of ILK was positively correlated with the expressions of α-SMA and FN and negatively correlated with E-cadherin expression.
Conclusion

IL-1β induces TEMT partly depending on TGF-β1/ILK signal way, partly via which emodin inhibits the TEMT induced by IL-1β.

Key words: Interleukin-lβ, Emodin, Transforming growth factor-betal, Integrin-linked kinase, Rats

Figure 1

Effects of IL-1β, emodin and SB431542 on morphological changes of NRK52E cells (Inverted phase contrast microscopy, ×40) A: Blank control group; B: Emodin control group; C: IL-1β-induced group; D: Emodin-inhibited group; E: SB431542-inhibited group; F: Emodin plus SB431542-inhibited group; G: Emodin-pretreated group; H: Emodin-reversed group."

Table 1

Effects of emodin, SB431542 and IL-1β on the secretion of FN and expressions of α-SMA,E-cadherin, TGF-β1 and ILK in NRK52E cells (x±s )"

Group n α-SMA E-cadherin FN (mg/L) TGF-β1 ILK
Blank control 6 65.47±1.73 82.49±1.04 54.65±3.09 7.04±1.19 36.08±3.08
Emodin control 6 64.87±2.05 81.62±2.09 54.64±2.35 6.85±1.23 36.16±2.72
IL-1β-induced 6 140.43±3.05* 36.04±2.82* 124.78±3.20* 19.16±2.12* 82.37±1.25*
Emodin-inhibited 6 95.77±2.12* 63.83±1.24* 81.54±2.41* 11.12±1.46* 55.34±1.65*
SB431542-inhibited 6 86.38±2.61*△□ 69.61±1.35*△□ 75.60±2.09*△□ 19.55±2.03*□ 50.34±1.53*△□
Emodin plus SB431542-inhibited 6 80.67±2.41*△▲□ 72.62±2.92*△▲□ 68.69±2.20*△▲□ 10.95±1.87*△▲ 44.92±2.24*△▲□
Emodin-pretreated 6 95.75±2.80* 65.41±2.64* 81.61±2.17* 10.73±1.56* 61.30±2.69*
Emodin-reversed 6 140.20±3.29*□ 36.33±2.74* 122.40±2.24* 18.56±1.94*□ 83.32±1.18*□

Table 2

Correlation analysis between expressions of TGF-β1, ILK and expressions of α-SMA, E-cadherin and secretion of FN"

Spearman correlation (r value/P value) Index
α-SMA E-cadherin FN ILK
TGF-β1 0.780/0.020 –0.760/0.022 0.800/0.018
ILK 0.993/0.003 –0.987/0.001 0.991/0.000 0.980/0.001
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