Effects of Huihui Gansong Yin on Proliferation of Rat Glomerular Mesangial Cells,Cell Cycle and Expression of FN,Col1α1 and Col4α1 mRNA

Objective: To investigate the effects of Huihui Gansong Yin( HGY) on the proliferation of rat glomerular mesangial cells( MCs),cell cycle,fibronectin( FN),collagen type Ⅰ alpha 1( Col1α1) and collagen typeⅣ alpha 1( Col4αl) induced by high glucose. Method: The serum containing HGY was prepared by adopting the method of serum pharmacology of TCM. Rats were divided into the blank group( distilled water,10 mL·kg- 1),glurenorm group( 10 mg·kg- 1),HGY group( 5,10 g·kg- 1). There were 10 rats in each group. At the end of the 3thday serum was obtained. HBZY-1 cells( MCs) were divided into 5 groups: 10%blank serum group,high-glucose + 10% blank serum group,high-glucose + 10% serum containing glurenorm10 mg·kg- 1,high-glucose + 10% serum containing HGY 10,5 g·kg- 1. MCs were cultured in 96-well plates( 2 ×104/ mL). After 24,48,72 h incubation with above indicated administration,the CCK-8 assay was used as a qualitative index of cell proliferation. Another MCs were cultured in 25 cm2cell culture bottles( 2 × 104/ mL).The cells were harvested 48h after treatment. Cell cycle analysis was performed by flow cytometry. FN,Col1α1and Col4α1 mRNA were measured using real-time PCR. Result: Compared with 10% blank serum group,highglucose( 30 mmol·L- 1) induced obvious proliferation of the MCs( P 0. 01). High-glucose induced decrease of cells in G0/ G1phase( P 0. 01),but increased cells in S phase( P 0. 01). High-glucose induced high levels of the expression of FN,Col1α1 and Col4α1 mRNA in MCs( P 0. 01). The above indicated administration,no matter for 48 h or 72 h,suppressed high glucose induced MCs proliferation and arrested with the cell cycle at G1/S phase,meanwhile,attenuated the expression of FN,Col1α1 and Col4α1 mRNA in MCs induced by high glucose( P 0. 01 or P 0. 05). Conclusion: The serum containing HGY could arrest with the cell cycle at G1/ S phase and inhibit the proliferation of MCs induced by high glucose. The mechanism is likely to be related that HGY suppressed high glucose induced expression of FN,Col1α1 and Col4α1 mRNA in rat mesangial cells.