Qingrejiedu Ziyinqingre,, Blood Circulation Of T2dm Pi3k Signaling Transduction Research

1.PurposeChinese traditional medicine has a good effect on treating diabetes mellitus whether in changing the symptom or preventing the neopathy. Especially it has predominance that it treats the diabetes as a whole and tries to find the root cause.Our project is to compare the treating effect of "QINREJIEDU","ZIYINQINGRE" and "HUOXUEHUAYU" on type 2 diabetes mellitus and its vascular pathological changes and on the insulin PI3K signal transduction of skeletal muscle cells according to the theory of "BIANZHESGLUNZHI" of chinese traditional medicine.We also observe and compare the effect of "QINREJIEDU","ZIYINQINGRE" and "HUOXUEHUAYU" on avoirdupois,blood sugar and insulin;on No in serum and ET;on ultra-structure of aorta abdominals;on the insulin PI3K signal transduction of skeletal muscle cells of type 2 diabetes mellitus rat. We aim to explore the treating essence of "QINREJIEDU","ZIYINQINGRE" and "HUOXUEHUAYU" on diabetes mellitus and its vascular pathological changes and explore the action of "REDU" in causing diabetes mellitus and its neopathy by investigate the signal transduction.The final purpose is to supply a new path to explore the treatment of diabetes mellitus by the Chinese traditional medicine.2.Method2.1 Theory researchReview the literature about the epidemiology,pathogeny,diagnosis, treatment and mechanism of neopathy of the diabetes mellitus and also review the literature about the animal model of type 2 diabetes mellitus. We also review the literature about pathogeny,"BIANZHENGLUNZHI", treatment and "FANGYAO" of diabetes mellitus in the Chinese traditional medicine.2.2 Experimental researches (1)Select 7 rats as normal group from 84 wistar rats;other 77 rats are fed by foods with high quantity of heat,high sugar,and high fattiness and with injection of STZ into the abdominal cavity to build the animal model.Then the 77 rats are grouped randomly into the model group, Gliclazide group,and groups with high,middle and low dose of herb portentillae respectively,groups with high,middle and low dose of radices salvia miltiorrhizae respectively and groups with high,middle and low dose of metropolis respectively.There are 7 rats in each group. When finish building the model,food is changed into normal condition and treats the rats with corresponding medicine for 4 weeks.(2)We weigh up the weight of the rats before and after the treatment respectively.(3)We collect the peripheral blood before and after the treatment respectively,and measure the blood sugar using Roche ACCU-CHEK Advantage blood glucose meter.(4)We collect the venues blood before and after the treatment respectively,and measure the blood insulin using radio-immunity.(5)We collect the venues blood before and after the treatment respectively and measure the blood nitric oxide using nitrate reeducates.(6)We collect the venues blood before and after the treatment respectively,and measure the blood ET.(7)After the experiment,we kill the rats and take out the aorta abdominals to make slices,then use uranyl acetate-lead citrate double staining and observe the ultra-structure of aorta abdominals with electron microscope.(8)After the experiment,we kill the rats and take out the abdominal muscle to make slices,then use uranyl acetate-lead citrate double staining and observe the ultra-structure of skeletal cells with electron microscope.(9)After the experiment,we kill the rats and take out the abdominal muscle to make slices,then use protein SDS-PAGE electrophoresis,immune marking to measure the expression of PI3K,PDK1 and GluT4ul in skeletal cells.3.Results(1)Theory research:the traditional chines medicine considers XIAOKE is caused by decreasing of JINYE;the neopathy is caused by ZAOREZHISHENG. On the other hand,treating the diabetes mellitus by "QINGREJIEDU" is proved in clinic practice.(2)Affection of "QINREJIEDU","ZIYINQINGRE" and "HUOXUEHUAYU" to the sugar metabolism of type 2 diabetes mellitus:the weight of rats in groups with high and middle dose of herb portentillae,groups with high and middle dose of radices salvia miltiorrhizae and groups with high, middle and low dose of metropolis increase very obviously(P＜0.01);the weight of rats in groups with low dose of herb portentillae,groups with low dose of radices salvia miltiorrhizae increase obviously(P＜0.05). Blood sugar in group with high dose of herb portentillae decreases obviously after the treatment(P＜0.05);whereas decreases very obviously(P＜0.01)in groups with middle and low dose of herb portentillae. Blood sugar in group with high,middle and low dose of radices salvia miltiorrhizae decrease but not obviously(P＞0.05).Blood sugar in groups with high and middle dose of metropolis decreases very obviously(P＜0.01); whereas decreases obviously(P＜0.05)in group with low dose of metropolis. INS in serum decrease very obviously(P＜0.01)in groups with high,middle and low dose of herb portentillae.INS in serum decrease very obviously(P＜0.01)in groups with high,middle and low dose of radices salvia miltiorrhizae.INS in serum decrease very obviously(P＜0.01)in group with high dose of metropolis;decrease obviously(P＜0.05)in group with middle and low dose of metropolis.(3)Affection of "QINREJIEDU","ZIYINQINGRE" and "HUOXUEHUAYU" to the active factors in blood vessels of type 2 diabetes mellitus:herb portentillae,radices salvia miltiorrhizae and metropolis can all decrease the blood nitric oxide,and they have similar intensity (according to the intensity of middle dose group of them)(P＜0.05).The intensity is similar to that of Gliclazide(compare the intensity of middle dose group of them with intensity of Gliclazide)(P＜0.05).Among them the affection of radices salvia miltiorrhizae is the best,metropolis is better and herb portentillae is the last one.Among all the groups of the three medicines,high dose groups are the best where as low dose groups are the worst,but there are no obvious differences among the corresponding groups of the three medicines.All of them can decrease the ET in serum and the intensity of herb portentillae and metropolis are similar(according to the intensity of middle dose group of them)(P＜0.05),and this intensity is similar to that of Gliclazide(compare the intensity of middle dose group of them with intensity of Gliclazide)(P＜0.05);the intensity of radices salvia miltiorrhizae is much weaker than that of herb portentillae and metropolis(according to the intensity of middle dose group of them)(P＜0.01),and also intensity of Gliclazide(P＜0.01).Among all the groups of the three medicines,high dose groups are the best where as low dose groups are the worst;there are no obvious difference between the intensity of large and middle dose groups of them,and no obvious difference between the intensity of middle and low dose groups(P＞0.05).(4)Affection of "QINREJIEDU","ZIYINQINGRE" and "HUOXUEHUAYU" to the ultra-structure of aorta abdominals of type 2 diabetes mellitus rat:the VEC structure of aorta abdominals is damaged seriously in model group. The permeation of aorta abdominals increases,the cells swell and break off;VSMC increases and moves to the theca internal,collage nous fibers increase,wall of aorta abdominal is becomes sclerous,and mitochondrion's also swell.The VEC and VSMC damage in aorta abdominals decrease in group of Gliclazide,herb portentillae,radices salvia miltiorrhizae and metropolis;among them,the affection of groups with high dose of herb portentillae,radices salvia miltiorrhizae and metropolis are the best whereas low dose groups are the worst.But there are no obvious differences among the corresponding groups of the three medicines.(5)Affection of "QINREJIEDU","ZIYINQINGRE" and "HUOXUEHUAYU" to the ultrastructure of skeletal muscle cells of type 2 diabetes mellitus rat:the ultrastructure of skeletal muscle cells damage seriously in model group because the collage nous fibers arrange in disorder,structure of sarcomere is not complete and looks indistinct;some mitochondrion's swell, the ridge of mitochondrion arranges in disorder or swell;glycogen increase obviously and has much fat;there is many increasing collage nous fibers in the mesenchyma.Compare to the model group,the damage decreases in the groups of herb portentillae,radices salvia miltiorrhizae and metropolis and among them affection of the high dose group is best,the middle dose group is better and the low dose group is the worst;the affection of the corresponding group between herb portentillae and metropolis is similar and better than radices salvia miltiorrhizae.(6)Affection of "QINREJIEDU","ZIYINQINGRE" and "HUOXUEHUAYU" to the PI3K signal transduction of skeletal muscle cells of type 2 diabetes mellitus rat:the expression of PI3K in model group is much lower than normal group(P＜0.05).The expression in group of Gliclazide is a little bit lower than normal group but there is no obvious difference(P＞0.05).It is much higher than model group(P＜0.05).All the herb portentillae groups are higher than normal group especially the high dose group(P＜0.05),but there is no obvious difference between the middle,low dose group and the normal group(P＞0.05).All the herb portentillae groups are higher than model group(P＜0.01).Among them the affection of the high dose group is best,the middle dose group is better and the low dose group is the worst,and compare to the low dose group the high dose group has obvious difference(P＜0.05).All the radices salvia miltiorrhizae groups are lower than normal group(P＜0.05)but only a little bit lower than model group and has no statistics difference(P＞0.05).All the metropolis groups are higher than normal group especially the high dose group(P＜0.05),but there is no obvious difference between the middle,low dose group and the normal group(P＞0.05).All the metropolis groups are higher than model group(P＜0.01).Among them the affection of the high dose group is best,the middle dose group is better and the low dose group is the worst,but compare to each other there is no obvious difference(P＞0.05). All the herb portentillae groups are higher than all the metropolis groups but there is no obvious difference(P＞0.05).The middle dose groups of metropolis and herb portentillae are higher than group of Gliclazide but there is no obvious difference(P＞0.05).All the metropolis groups and herb portentillae groups are higher than groups of radices salvia miltiorrhizae,among them the high and middle dose group of herb portentillae and the high dose group of metropolis are much higher than that of groups of radices salvia miltiorrhizae(P＜0.01)and other groups are higher than that of groups of radices salvia miltiorrhizae(P＜0.05).The middle dose group of radices salvia miltiorrhizae is lower than group of Gliclazide(P＜0.05).The expression of PDK1 in model group is lower than normal group(P＜0.05).Group of Gliclazide is a little bit lower than normal group but there is no obvious difference(P＞0.05).All the herb portentillae groups are higher than normal group but there is no obvious difference(P＞0.05).All the herb portentillae groups are much higher than model group(P＜0.01).Among them the affection of the high dose group is best,the middle dose group is better and the low dose group is the worst,but compare to each other there is no obvious difference(P＞0.05).All the radices salvia miltiorrhizae groups are lower than normal group(P＜0.05);its high dose group is higher than model group and the middle and low dose groups are lower than model group but there is no obvious difference(P＞0.05);and compare to each other there is no obvious difference(P＞0.05).All the metropolis groups are higher than normal group but there is no obvious difference(P＞0.05);Among them the affection of the high dose group is best,the middle dose group is better and the low dose group is the worst,but compare to each other there is no obvious difference(P＞0.05).All the herb portentillae groups are higher than all the metropolis groups but there is no obvious difference(P＞0.05);both of the middle dose group are higher than group of Gliclazide especially the middle dose group of herb portentillae and has statistics difference(P＜0.05),but the middle dose group of metropolis has no obvious difference(P＞0.05);All the groups of herb portentillae and metropolis are higher than the groups of radices salvia miltiorrhizae(P＜0.01).The middle dose group of radices salvia miltiorrhizae is lower than group of Gliclazide but there is no obvious difference(P＞0.05).The expression of GluT4 in model group is much lower than modal group(P＜0.05).The expression in group of Gliclazide is a little bit lower than normal group but there is no obvious difference(P＞0.05).It is much higher than model group(P＜0.05).The high dose group herb portentillae is a little bit higher than normal group and the middle,low dose group is lower than the normal group but there is no obvious difference(P＞0.05). All the herb portentillae groups are higher than model group(P＜0.05). Among them the affection of the high dose group is best,the middle dose group is better and the low dose group is the worst,but compare to each other there is no obvious difference(P＞0.05).All the radices salvia miltiorrhizae groups are lower than normal group but there is no obvious difference(P＞0.05).All the radices salvia miltiorrhizae groups are a little bit higher than model group but has no statistics difference(P＞0.05);and compare to each other there is no obvious difference(P＞0.05). The high dose group of metropolis is a little bit higher than normal group and the middle and low dose group is lower than the normal group but there is no obvious difference(P＞0.05).Among them the affection of the high dose group is best,the middle dose group is better and the low dose group is the worst,but compare to each other there is no obvious difference(P＞0.05).All the herb portentillae groups are higher than all the metropolis groups but there is no obvious difference(P＞0.05).The middle dose groups of metropolis and herb portentillae are higher than group of Gliclazide but there is no obvious difference(P＞0.05).All the metropolis groups and herb portentillae groups are higher than groups of radices salvia miltiorrhizae,among them the high dose group of herb portentillae and metropolis are much higher than that of groups of radices salvia miltiorrhizae(P＜0.05)and other groups have no statistic difference(P＞0.05).The middle dose group of radices salvia miltiorrhizae is lower than group of Gliclazide but there is no obvious difference(P＞0.05).4.Conclusion(1)"REDU" is the basic cause and the important mechanism of DM and its neopathy.(2)Both the "QINREJIEDU" and "ZIYINQINGRE" can decrease the blood sugar effectively and they have similar intensity,but "QINREJIEDU" is a little bit better than "ZIYINQINGRE".The possible mechanism of "QINREJIEDU" is:①protect the B cells of islet of pancreas and increase the secretion of INS;②up regulate the signal transduction of insulin in insulin sensitive skeletal muscle cells and increase the movement of blood sugar into skeletal muscle cells.The possible mechanism of metropolis to decrease the blood sugar is:①increase the secretion of INS;②up regulate the signal transduction of insulin in insulin sensitive skeletal muscle cells and increase the movement of blood sugar into skeletal muscle cells."HUOXUEHUAYU" cannot increase the secretion of INS and up regulate the signal transduction of insulin in insulin sensitive skeletal muscle cells,so it cannot control the blood sugar of diabetes mellitus rat. (3)"QINREJIEDU","ZIYINQINGRE" and "HUOXUEHUAYU" can decrease the VEC damage of type 2 diabetes mellitus rat and the increasing of VSMC; "QINREJIEDU" and "ZIYINQINGRE" have similar intensity and better than "HUOXUEHUAYU".The possible mechanism to prevent the pathology changes of large blood vessel is:①decrease the level of blood sugar to prevent its damage to VEC and VSMC;②decrease the secretion of ET by VEC and decrease its damage to VEC and VSMC;③decrease the expression of iNOS of VSMC and down-regulate the production of nitric oxide to prevent its cellular poison.The possible mechanism of "QINREJIEDU" to prevent the pathology changes of large blood vessel is:①decrease the secretion of ET by VEC and decrease its damage to VEC and VSMC;②decrease the expression of iNOS of VSMC and down-regulate the production of nitric oxide to prevent its cellular poison.(4)"QINREJIEDU","ZIYINQINGRE" and "HUOXUEHUAYU" can protect the skeletal muscle and decrease the damage of it;"QINREJIEDU" and "ZIYINQINGRE" have similar intensity and are better than "HUOXUEHUAYU". The possible mechanism to protect the skeletal muscle by "QINREJIEDU" and "ZIYINQINGRE" is:①increase the secretion of INS to decrease the blood sugar level②up regulate the signal transduction of insulin in insulin sensitive skeletal muscle cells and increase the movement of blood sugar into skeletal muscle cells.The possible mechanism to protect the skeletal muscle by "HUOXUEHUAYU" is to eliminate the free radical and decrease the cellular poison of ROS by the salvianolic acid compound.