高糖诱导人肾间质成纤维细胞c-met的表达及黄芪对其的调节作用

doi:10.3736/jcim20080510

Abstract

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

Shan Mou, Zhao-hui Ni, Qing-yi Zhang. Expression of c-met in human kidney fibroblasts induced by high glucose in vitro and the regulation of Radix Astragali. Journal of Chinese Integrative Medicine, 2008, 6(5): 482-487.

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URL: http://www.jcimjournal.com/EN/10.3736/jcim20080510

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References 12

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Group	n	c-met mRNA
Group	n	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

Group	n	c-met protein
Group	n	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

Group	n	TGF-β1 mRNA
Group	n	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^△△

Group	n	c-met protein		c-met mRNA
Group	n	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^△△▲

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

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