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Journal of Chinese Integrative Medicine ›› 2008, Vol. 6 ›› Issue (5): 482-487.doi: 10.3736/jcim20080510

• Original Experimental Research • Previous Articles     Next Articles

Expression of c-met in human kidney fibroblasts induced by high glucose in vitro and the regulation of Radix Astragali

Shan Mou(), Zhao-hui Ni, Qing-yi Zhang   

  1. Department of Nephrology, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China
  • Received:2008-03-14 Online:2008-05-20 Published:2008-05-15
  • Contact: MOU Shan E-mail:shan_mou@hotmail.com

Objective: To investigate the effect of high glucose on the expression of c-met in human kidney fibroblasts in vitro, and to explore the regulation of Radix Astragali.
Methods: A cell culture system of human kidney fibroblasts was developed in vitro. The human kidney fibroblasts were divided into normal control group, high glucose group and mannitol group. Expressions of c-met and transforming growth factor-beta1 (TGF-β1) mRNAs were detected by reverse transcription polymerase chain reaction (RT-PCR) and the expressions of c-met protein were analyzed by Western blot method after 6-, 12-, 24-, 48- and 96-hour culture. The human kidney fibroblasts were also cultured with 10 % Radix Astragali containing serum; the expressions of c-met mRNA and protein were detected after 24- and 48-hour culture.
Results: Compared with the normal control group, expression of c-met mRNA in the high glucose group was significantly increased after 12-hour culture (P<0.05), arriving at the peak after 24-hour culture (P<0.01). The level of TGF-β1 mRNA was higher in the high glucose group than that in the normal control group after 24-hour culture (P<0.05), arriving at the peak after 96-hour culture (P<0.01). Forty-eight hours after treating with 10% Radix Astragali containing serum, the levels of c-met mRNA and protein in fibroblasts were increased, and were higher than those in the high glucose group (P<0.01, P<0.05).
Conclusion: High glucose can induce the expressions of c-met mRNA and protein in earlier period, and then inhibit the expressions. Radix Astragali can up-regulate the expressions of c-met mRNA and protein of human kidney fibroblasts, which may be one of its action mechanisms in delaying the progression of diabetic nephropathy.

Key words: proto-oncogene protein c-met, diabetic nephropathy, Radix Astragali, fibroblasts, transforming growth factor-betal

CLC Number: 

  • R285.5

Figure 1

Expression of c-met mRNA after 24-hour culture in different groups 1: Normal control group; 2: High glucose group; 3: Mannitol group."

Table 1

Effect of high glucose on expression of c-met mRNA ($\bar{x}$±s, %)"

Group n c-met mRNA
6 hours 12 hours 24 hours 48 hours 96 hours
Normal control 6 29.57±1.17 29.40±3.80 29.87±5.05 32.23±3.96 28.33±2.37
High glucose 6 32.10±2.95 48.80±7.37* 80.63±5.00** 55.63±5.05* 30.63±5.05
Mannitol 6 29.50±3.70 41.47±9.33 63.37±6.01**△ 42.83±9.15 34.43±6.82

Table 2

Effect of high glucose on expression of c-met protein ($\bar{x}$±s)"

Group n c-met protein
6 hours 12 hours 24 hours 48 hours 96 hours
Normal control 6 1.04±0.18 1.06±0.12 1.03±0.16 1.00±0.00 1.04±0.05
High glucose 6 1.38±0.18 1.53±0.16* 4.99±0.18** 1.62±0.18* 1.32±0.19
Mannitol 6 1.04±0.19 1.10±0.11 0.99±0.21△△ 0.94±0.06△△ 1.06±0.17

Figure 2

Expression of c-met protein after 24-hour culture in different groups 1: Normal control group; 2: High glucose group; 3: Mannitol group."

Table 3

Effect of high glucose on expression of TGF-β1 mRNA ($\bar{x}$±s, %)"

Group n TGF-β1 mRNA
6 hours 12 hours 24 hours 48 hours 96 hours
Normal control 6 28.23±2.63 31.27±7.27 38.03±4.51 40.47±4.45 43.57±3.42
High glucose 6 28.73±5.77 40.50±1.83 62.60±3.28** 85.87±5.64** 103.73±6.21**
Mannitol 6 26.30±4.26 36.83±3.90 50.03±3.69* 55.53±5.75*△△ 56.57±10.43△△

Figure 3

Expression of c-met mRNA after treating with 10% Radix Astragali serum for 48 hours 1: High glucose group; 2: 10% control serum group; 3: 10% Radix Astragali serum group."

Figure 4

Expression of c-met protein after treating with 10% Radix Astragali serum for 48 hours 1: High glucose group; 2: 10% control serum group; 3: 10% Radix Astragali serum group."

Table 4

Expressions of c-met protein and mRNA in the fibroblasts treating with 10% Radix Astragali serum ($\bar{x}$±s, %)"

Group n c-met protein c-met mRNA
24 hours 48 hours 24 hours 48 hours
High glucose 6 1.01±0.01 1.00±0.02 83.03±4.35 54.43±5.35
10% control serum 6 1.08±0.11 1.14±0.19 81.80±10.71 60.33±6.02
10% Radix Astragali serum 6 1.29±0.20 3.37±0.48△▲ 85.77±5.19 83.60±4.25△△▲
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