Pharmacodynamics Study On Antitumor Effective Constituents Of Juglans Mandshurica Maxim Husk Extraction And Its Mechanism

Objective To research the pharmacodynamics and its related mechanisrn on antitumor constituents of Juglans mandshurica Maxim husk extraction.Promoting futher processing and exploiting Juglans mandshurica Maxim husk.Providing relevant theotrtical basis for development medical value.MethodsTest one:First,By MTT method,We obseved the antitumor effect of extration in vitro.Second,By the methods of MTT and ELISA,detecting inhibition of the SGC7901,HepG2 and SW620 cell proliferation and v-Src protein expression.Test two:First,using ELISA method,we detected protein expression of p38MAPK in the extracellular fluid of SGC7901,HepG2 and SW620.Second,using ELISA method,we detected protein expression of matrix metalloproteinase-2(MMP-2)and metalloproteinase-9(MMP-9)in the extracellular fluid of SGC7901,HepG2 and SW620.ResultsFirst,compared with the control group,all the Juglans mandshurica Maxim husk extractions can inhibit tumor cells proliferation,The petroleum ether extraction has the most effect,IC50 of HepG2,SGC7901 and SW620 cells were 123.82 ug/mL,28.39 mg/mL and 59.54 mg/mL.Petroleum ether extraction's IC50 is half of the positive drug,significant inhibition of SGC7901 cell proliferation.Second,the inhibition to HepG2 cell's proliferation by pinostrobin,emodin,5-hydroxy-7,4'-dimethoxyflavone is in turn increases.The IC50 of emodin and 5-hydroxy-7,4'-dimethoxyflavone are 63.52?g/mL,46.66?g/mL,they are more effective than Petroleum ether extraction.p-Sitosterol,Juglone,stigmasterol extracted parts is invalid;IC50 of inhibition SGC7901 cell proliferation lead small to large for emodin,pinostrobin,5-hydroxy-7,4'-dimethoxyflavone,?-Sitosterol,Juglone,stigmasterol.IC50 of emodin and 5-hydroxy-7,4'-dimethoxyflavone inhibition SW620 cell proliferation are 40.95?g/mL,7.71?g/mL,effection of anti-cancer better than petroleum ether extraction.Third,emodin and 5-hydroxy-7,4'-dimethoxy flavone inhibited v-Src protein expression in supernatant fluid of HepG2,SGC7901,SW620 showed a concentration-dependent.High concentration group more effective.In addition to the low concentration group of SW620,compared with the control group,the other groups of emodin were significant(P<0.05).5-hydroxy-7,4'-dimethoxyflavone all groups in HepG2 cells,hight and middle concentration groups are statistically significant(P<0.05,P<0.01).SW620,SGC7901 in the high centrations of pinostrobin group,HepG2 in the middle the v-src protein expression was inhibited(P<0.05).Forth,detected effection of emodin on p3 8MAPK protein expression by immunocytochemistry method.Compared with the control group,emodin can inhibit the expression of p38MAPK protein.Cells claybank particle reduce,color shallow.5-hydroxy-7,4'-dimethoxyflavone made the p38 MAPK protein color in the HepG2 and SW620 cell shallow,no obvious change in the SGC7901 cell.Fifth,Compared with the control group,emodin can inhibit the expression of MMP-2 protein(P<0.05),Emodin low concentration group can restrain about 50%MMP-2 protein expression on SW620 cells,HepG2 cells with high emodin concentration,SW620 cells with high and middle emodin concentration are statistically significan(P<0.01).Compared with the control group,high concentration of 5-hydroxy-7,4'-dimethoxyflavone role in SGC7901 and SW620 to reduce the expression of protein(P<0.05).there are no effect.Sixth,compared with the control group,Emodin can reduce the protein of MMP-9 expression(P<0.05).In addition to high concentration of 5-hydroxy-7,4'-dimethoxyflavone have effect to protein MMP-9 expression of HepG2 others haven't.ConclusionFirst,petroleum ether extraction has the best effect on HepG2,SGC7901 and SW620 cell proliferation inhibition.Second,pinostrobin,emodin,5-hydroxy-7,4'-dimethoxyflavone can inhibit cell preliferation,showed a concentration-dependent,emodin has the best effect.Third,5-hydroxy-7,4'-dimethoxyflavone has better effect on inhibition v-Src protein expression of HepG2 and SW620 cell than emodin,instead emodin is better to inhibit cell proliferation.5-hydroxy-7,4'-dimethoxyflavone should through the other way or joint effect to inhibit tumor proliferation.Fourth,emodin can inhibit the expression of p38MAPK protein.5-hydroxy-7,4'-dimethoxyflavone can inhibit the expression of p38MAPK protein in the cell of HepG2?SW620,but can't inhibit the expression in SGC7901 cell.Fifth,emodin can inhibit the expresstion of MMP-9 and MMP-2;5-hydroxy-7,4'-dimethoxyflavone inhibit the expresstion of MMP-2 in SGC7901 cell and don't affect the protein expression of the other tumor cells.