Herbal Drugs Attenuated DN By ERK And TGF-β-signaling Pathway

In the process of our people battle with disease for thousands years of history, by practice, constant awareness gradually accumulated a wealth of herbal knowledge. Chinese herba have unique advantages on lasting effects and little side effect. The efficacy of many traditional Chinese medicine not only withstood the test of long-term medical practice, but also has been confirmed by Sciences. However, due to the complexity of the active ingredients of traditional Chinese medicine and doubt the efficacy of Chinese herbal medicine, and mechanisms of many Chinese medicinal has not yet been completely understanded, resulting in prevented the extensive promotion and application . Therefore, it is extremely urgent to in-depth study on Chinese medicinal active ingredient and to explore its mechanism.In this paper, screening effective herbal medicines from nine kinds of blood circulation medicines in the cellular level, and then observed the effective of Chinese herbal on attenuated diabetic nephropathy in rats by ERK,TGF-β.After process of boiling, concentrated and freeze-dried ,ultimately, gaining the freeze-dried powder of nine kinds of medicinal plants;In vitro1) MTT assay: nine blood circulation medicines of water extracts(except Giant Knotweed Rhizome) at lmg/ml have no effects on cell proliferation, so can determine the scope of drug treatmentin the follow-up experiments;2) Establishing cell models of over-expression extracellular matrix by add the rhTGF-β1;3) Use 2) have establishment of the model, treating a variety of blood circulation herbal water extracts (final concentration 200μg/ml)to cells, and then observating the expression of p-Smad2, and found Lycopus can significantly reversed the increasing p-Smad2 protein that cause by rhTGF-β1 stimulated, while the expression of Smad2 did not affect;4) Specific studies on water extract of Lycopus affected on TβRⅠ, TβRⅡ, p-Smad2, Smad2, Smad4, Smad7, p-ERK1/2 and ERK1/2 protein expression, the study confirmed that rhTGF-βlean significantly increasing expression. In the latter experiments ,finding Lycopus can reverse the increasing expression of TβR-Ⅱ, p-Smad2, Smad4, p-ERK1/2 protein,but have no effects on Smad2,TβRⅠand ERK1/2 and reduce Smad7.三) In vivo1) Successfully established a rat model of diabetic nephropathy experimental, that can provided a guarantee for the follow-up studies;2) Although not lowering blood sugar, gaining the weight and decreased urine volume, treatment group of water extract of Lycopus can significantly reducing the STZ rats of SCr, BUN, urinary protein (P<0.05), and increased SOD activity (P<0.05); 3)The reslut of RT-PCR show that Lycopus in the dose gropu (6g/kg/day) can significantly decreased the mRNA of TGF-β1 in the STZ rat; Western blot (WB) test results showed that all Lycopus treatment group can significantly reduced p-Smad2 protein expression (P<0.01), but no effect on Smad2 protein expression.