糖肾宝含药血清对高糖培养大鼠系膜细胞细胞外调节蛋白激酶1/2磷酸化及转化生长因子-β1、单核细胞趋化蛋白-1表达的影响

Effect of Serum Containing Tangshenbao on Phosphorylation of Extracellular Regulated Protein Kinases 1/2 and Expression of Transforming Growth Factor-β1 and Monocyte Chemoattractant Protein-1 in Rat Mesangial Cells Induced by High Glucose

【目的】探讨中药复方糖肾宝(主要由黄芪、大火草、葛根、莪术、三七组成)对高糖培养的大鼠系膜细胞(MCs)细胞外调节蛋白激酶1/2磷酸化(pERK1/2)及转化生长因子-β1(TGF-β1)、单核细胞趋化蛋白-1(MCP-1)表达的影响。【方法】体外培养大鼠MCs,随机分为正常组,高糖对照组,糖肾宝低、中、高剂量组(剂量分别为13.5、27、54 g/kg),厄贝沙坦组(剂量为25 mg/kg)。除正常组外,其余各组分别予以30 mmol/L葡萄糖+正常大鼠血清,糖肾宝低、中、高剂量血清,厄贝沙坦血清进行干预48 h。采用Western blot法检测细胞中pERK1/2水平、TGF-β1蛋白变化,RT-PCR法检测TGF-β1、MCP-1 mRNA表达,酶联免疫吸附法检测MCP-1蛋白和纤维连接蛋白(FN)表达。【结果】高糖对照组pERK1/2水平、TGF-β1、MCP-1、FN表达均显著高于正常组(P<0.05或P<0.01);药物干预的各组pERK1/2水平、TGF-β1、MCP-1、FN表达均显著低于高糖对照组(P<0.05或P<0.01);糖肾宝高剂量组pERK1/2水平显著低于厄贝沙坦组(P<0.01),TGF-β1、MCP-1、FN表达与厄贝沙坦组比较差异无统计学意义(P>0.05)。【结论】中药复方糖肾宝能抑制高糖培养的MCs细胞外基质的分泌,其机制可能与抑制ERK1/2磷酸化,并进而抑制TGF-β1和MCP-1表达有关。

Objective To investigate the effect of Tangshenbao Granula, a Chinese herbal recipe mainly composed of Radix Astragali, root of Anemone tomentosa （Maxim.） Pei, Radix Puerariae, Rhizoma Curcumae, and Radix Notoginseng, on the phosphorylation of extracellular regulated protein kinases 1/2 （pERK1/2） and the expression of transforming growth factor-β1 （TGF-β1） and monocyte chemoattractant protein-1 （MCP-1） in rat mesangial cells （MCs） induced by high glucose. Methods The rat MCs were randomly divided into normal group, high glucose control group, irbesartan group （in the dosage of 25 mg/kg） , and low-, middle- and high-dose Tangshenbao groups （in the dosage of 13.5, 27, 54 g/kg, respectively） . The rats in the normal group were given normal rat serum alone, the other groups were all given 30 mmol/L glucose and normal rat serum, and additionally, Tangshenbao groups and irbesartan group were given serum containing low-, middle- and high-dose Tangshenbao and irbesartan, respectively. The expression of pERK1/2 and TGF-β1 protein was examined by Western blotting method. TGF-β1 and MCP-1 mRNA expression levels were observed by real-time polymerase chain reaction （PCR） . The expression of MCP-1 andfibronectin （FN） protein was examined by enzyme-linked immunosorbent assay. Results pERK1/2, TGF- 151, MCP-1 and FN expression levels in high glucose control group were significantly higher than those in the normal group （P〈0.05 or P〈0.01）, and were decreased in the medication groups （P〈0.05 or P〈0.01 compared with those in the high glucose control group） . The pERK1/2 level in high-dose Tangshenbao group was significantly lower than that in the irbesartan group （P〈0.05 or P〈0.01）, while TGF-β1, MCP-1 and FN expression levels were not significantly different from those in irbesartan group （P〉0.05） . Conclusion Tangshenbao can inhibit the secretion of extracellular matrix in MCs induced by high glucose, and its mechanism may be related to the inhibition of ERK phosphorylation and TGF-β1 and MCP-1 expression.