| Purposes:Through inject baicalin of different doses to human lung fibroblasts,observe baicalin changes in cell of normal on CTGF,FN,ILK,CollagenⅠand Ⅲ,and discuss the prevention mechanism of baicalin for huamn lung fibroblasts.Methods: After cultivate human lung fibroblasts,carry through passage and divide into five groups and bacteria is 2×105,the blank group,prednisone aoetate group,baicalin high dose group,baicalin dose group,baicalin low dose group,to inject medcine for every group;prednisone aoetate group add 1m M,and high dose group and low dose group and dose group of baicalin different add 10 m M,5m M,1m M;Culttivating on co2 incubator and continuous dosing in 24 h,48h,72h;Detecting cell proliferctim rate in MTT and drawing curve;Applying molecular biology techniques include RT-PCR and western blot detect the expression level of CTGF,FN,ILK,CollagenⅠand Ⅲ m RNA and protein.Result:1.MTT measure proliferation rate which baicalin intervene human lung fibroblasts HFL1 cell is interposed by different cell of concentration in 72 h.Cell proliferation is obviously.as a result of MTT measure,The role is of dependence of concentration-With increase of baicalin potency,OD value is become higher,which the way that potency depend on,cell prolifer become obvious when potency reach to 10 m M,compared with the control group,there is meaningful in statistics(P<0.05).2.Research that baicalin have influence on m RNA express of CTGF、FN、ILK、CollagenⅠ、Ⅲ for HLF12.1 Express of CTGF m RNA The results show that Compared prednisone acetate group with the blank group,prednisone acetate group is obviously decline in express level of CTGF m RNA HFL1 cell under interposed of baicalin and prednisone acetate.there is remarkable statistical significance(P<0.01);for interposed of baicalin,high dose group and dose group and low dose gruop is mean in express level of CTGF m RNA HFL1 cell.there is remarkable statistical significance(P<0.05),Compared baicalin high dose group with the blankgroup,there is not statistical significance(P>0.05),Compared high dose group with active control group there is remarkable statistical significance(P<0.05)Compared,dose group with low dose group,there is remarkable statistical significance(P<0.01).Compared dose group with with low dose group,without significance(P>0.05).Compared the blank group with low dose group and dose group without significance(P>0.05).2.2Express of FN m RNA Compared prednisone acetate group with negtive control group,active control group is obviously decline in express level of FN m RNA HFL1 cell under interposed of baicalin and prednisone acetate.Compared with negtive control group,there is remarkable statistical significance(P<0.01).baicalin high dose group and dose group and low dose group and the prednisone aoetate group,there is remarkable statistical significance(P<0.01).Compared with low dose group and dose group and high dose group(P<0.01);Compared low dose group with dose group,there is remarkable statistical significance(P<0.05).2.3Express of ILK m RNA Compared prednisone acetate group with negtive control group,prednisone acetate group is obviously decline in express level of ILK m RNA HFL1 cell under interposed of baicalin and prednisone acetate.Compared with the blank group,there is remarkable statistical significance(P<0.01).Compared dose group with low dose group,there is remarkable statistical significance(P<0.01).2.4Express of collagenⅠm RNA Compared prednisone acetate group with the blank group,prednisone acetate group is obviously decline in express level of collagenⅠ m RNA HFL1 cell under interposed of baicalin and prednisone acetate.Compared with the prednisone acetate group,baicalin high dose group and dose group and low dose group,collagenⅠpositive is expressed in group cell,there is remarkable statistical significance(P<0.01).Compared baicalin high dose group with prednisone acetate group,there is notstatistical significance(P>0.05),Compared dose group with active control group,there is remarkable statistical significance(P<0.01).Compared baicalin high dose group with low dose group,there is remarkable statistical significance(P<0.01).Compared low dose group with dose group,there is not statistical significance(P>0.05).2.5Express of collagen ⅢCompared prednisone acetate group with the blank group,prednisone acetate group is obviously decline in express level of collagen Ⅲ m RNA HFL1 cell under interposed of baicalin and prednisone acetate.Compared with negtive control group,there is remarkable statistical significance(P<0.01).Compared baicalin high dose group and dose group and low dose group with prednisone acetate group there is remarkable statistical significance(P<0.01).Compared high dose group with dose group,there is statistical significance(P<0.05).Compared dose group with low dose group,there is remarkable statistical significance(P<0.01).2.Research that baicalin have influence on protein express of CTGF、FN、ILK、CollagenⅠ、Ⅲ for HLF3.1Express of CTGF protein Compared prednisone acetate group with the blank group,prednisone acetate group is obviously decline in express level of CTGF protein HFL1 cell under interposed of baicalin and prednisone acetate.Compared with the blank group,there is remarkable statistical significance(P<0.01),compared baicalin high dose group and dose group and low dose group with prednisone acetate group,there is remarkable statistical significance(P<0.01).Compared high dose group with dose group,there is remarkable statistical significance(P<0.01),compared high dose group with low dose group,there is remarkable statistical significance(P<0.01),compared dose group with low dose group,there is remarkable statistical significance(P<0.01).3.2Express of ILk protein Compared prednisone acetate group with negtive control group,prednisone acetate group is obviously mean decline in express level of ILK protein HFL1 cell under interposed of baicalin and prednisone acetate.Compared with the blankgroup,baicalin high dose group and active control group,there is remarkable statistical significance(P<0.05).Compared low dose group with negtive control group,there is not statistical significance(P>0.05).Compared with prednisone acetate groupfor baicalin high dose group and dose group and low dose group,there is remarkable statistical significance(P<0.01).Compared high dose group with dose group,there is remarkable statistical significance(P<0.01),compared high dose group with low dose group,there is remarkable statistical significance(P<0.01),compared dose group with low dose group,there is not statistical significance(P>0.05).3.3Express of FN protein Compared prednisone acetate group with the blank group,prednisone acetate group is obviously mean decline in express level of FN protein HFL1 cell under interposed of baicalin and prednisone acetate.Compared with the blank group,baicalin high dose group and dose group and low dose group,there is remarkable statistical significance(P<0.01).Compared low dose group with the blank group,there is not statistical significance(P>0.05),compared with active control group,baicalin high dose group and dose group and low dose group,there is remarkable statistical significance(P<0.01),Compared high dose group with dose group,there is remarkable statistical significance(P<0.01),compared high dose group with low dose group,there is remarkable statistical significance(P<0.01),compared dose group with low dose group,there is remarkable statistical significance(P<0.01).3.4Express of CollagenⅠprotein Compared prednisone acetate group with the blank group,prednisone acetate group is obviously mean decline in express level of CollagenⅠ protein HFL1 cell under interposed of baicalin and prednisone acetate.Compared with the blank group,baicalin high dose group and dose group and low dose group,there is remarkable statistical significance(P<0.01).Compared baicalin high dose group and dose group and low dose group with prednisone acetate group,there is remarkable statistical significance(P<0.01),compared high dose group with low dose group,there is remarkable statistical significance(P<0.01),Compared high dose group with dose group,there is not statistical significance(P>0.05),compared dose group with low dosegroup,there is remarkable statistical significance(P<0.01).3.5Express of Collagen Ⅲ protein Compared prednisone acetate group with the blank group,prednisone acetate group is obviously mean decline in express level of CollagenⅢ protein HFL1 cell under interposed of baicalin and prednisone acetate.Compared with the blank group,there is remarkable statistical significance(P<0.01),compared high dose group with prednisone acetate group,there is not statistical significance(P>0.05),compared low dose group with prednisone acetate group,there is remarkable statistical significance(P<0.01),Compared high dose group with dose group,there is not statistical significance(P>0.05),compared high dose group with low dose group,there is statistical significance(P>0.01),compared dose group with low dose group,there is statistical significance(P>0.01).Conclusion:1 After HFL1 cell is controled by baicalin medcine in 72 h.Baicalin have control a role for cell factor on depth,with time of baicalin effect rise,change of cell proliferation is obvious and how much cell proliferation with close ties to cell factor.With regard to what baicalin can control some diease is that offer theoretical support of well.2 Compared with baicalin dose group and low dose group,m RNA control of baicalin high dose group is better for CTGF,FN,ILk,CollagenⅠⅢ,the same as prenisone aoetate group.baicalin may have better on lung disease.Baicalin high dose group is best,and nearly equal to prenisone aoetate group,take second place on baicalin dose group,the worst is baicalin low dose group.3 Compared with baicalin dose group and low dose group,protein control of baicalin high dose group is obvious better for CTGF,FN,ILk,CollagenⅠⅢ,the same as prenisone aoetate group.From express level of protein and above control of cell factor m RNA the view.In addition,prove that baicalin have a control better role for lung disease.,which is bad than prednisone acetate group. |
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