Effects of Huihui Gansong Yin on Proliferation of Rat Glomerular Mesangial Cells,Cell Cycle and Expression of FN, Col11 and Col41 mRNA

YUAN Ling; NAN Yi; WU You-xi; XU Jun; HUANG Xiu-lan

doi:10.11653/syfj2014040135

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Effects of Huihui Gansong Yin on Proliferation of Rat Glomerular Mesangial Cells,Cell Cycle and Expression of FN, Col11 and Col41 mRNA

更新时间：2024-01-04

- Effects of Huihui Gansong Yin on Proliferation of Rat Glomerular Mesangial Cells,Cell Cycle and Expression of FN, Col11 and Col41 mRNA
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 20, Issue 4, Pages: 135-140(2014)
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- DOI：10.11653/syfj2014040135
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- Published：2014
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YUAN Ling, NAN Yi, WU You-xi, et al. Effects of Huihui Gansong Yin on Proliferation of Rat Glomerular Mesangial Cells,Cell Cycle and Expression of FN, Col11 and Col41 mRNA[J]. Chinese journal of experimental traditional medical formulae, 2014, 20(4): 135-140.
DOI：

YUAN Ling, NAN Yi, WU You-xi, et al. Effects of Huihui Gansong Yin on Proliferation of Rat Glomerular Mesangial Cells,Cell Cycle and Expression of FN, Col11 and Col41 mRNA[J]. Chinese journal of experimental traditional medical formulae, 2014, 20(4): 135-140. DOI： 10.11653/syfj2014040135.

摘要

目的：探讨回回甘松饮（HGY）含药血清对高糖诱导的大鼠肾小球系膜细胞（MCs）增殖、细胞周期、纤维连接蛋白（FN）、Ⅰ型胶原α1多肽链（Col1α1）、Ⅳ型胶原α1多肽链（Col4α1）基因表达的影响。方法：采用中药血清药理学方法制备含药血清，将大鼠随机分为空白对照组（给予蒸馏水10 mL·kg-1）、格列喹酮（10 mg·kg-1）组、HGY（10，5 g·kg-1）组，每组各10只，每日灌胃给药2次，连续3 d后取大鼠血清。选取HBZY-1细胞株进行实验，共分为5组：10%空白血清组、高糖+10%空白血清组、高糖+10%格列喹酮含药血清组（10 mg·kg-1）、高糖+10%HGY含药血清组（10，5 g·kg-1）。细胞以2×104/mL接种于96孔细胞培养板上，分别于药物处理后24，48，72 h时，采用CCK-8法检测MCs增殖情况；细胞以2×104/mL接种于25 cm2细胞培养瓶中，于药物干预48 h时收集细胞，分别采用流式细胞术检测细胞周期变化，real-time PCR方法检测FN、Col1α1及Col4α1 mRNA的表达。结果：与10%空白血清组比较，高糖（30 mmol·L-1）能够诱导MCs增殖（P<0.01），并能使G0/G1期细胞比例减少（P<0.01），S期比例增加（P<0.01），能够使MCs中FN，Col1α1，Col4α1 mRNA表达量明显升高（P<0.01）。在上述含药血清干预的48，72 h时，各含药血清组MCs均受到一定程度抑制，发生G1/S期阻滞，并可降低高糖条件下MCs中FN，Col1α1，Col4α1 mRNA表达量，与高糖+10%空白血清组比较，具有显著性差异（P<0.01或P<0.05）。结论：HGY含药血清可使大鼠MCs在高糖环境下发生G1/S期阻滞，抑制其增殖，其机制可能与调控FN，Col1α1及Col4α1 mRNA表达有关。

Abstract

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 type Ⅳ 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 3th day 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 glurenorm 10 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

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 cm2 cell culture bottles(2×104/mL). The cells were harvested 48h after treatment. Cell cycle analysis was performed by flow cytometry. FN

Col1α1 and Col4α1 mRNA were measured using real-time PCR. Result: Compared with 10% blank serum group

high-glucose(30 mmol·L-1)induced obvious proliferation of the MCs(P<0.01). High-glucose induced decrease of cells in G0/G1 phase(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.

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