| ObjectivesImpaired glucose tolerance (IGT) is glycometabolism disorder between normal glucose tolerance (NGT) and diabetes mellitus (DM). The rate of IGT is higher than the rate of DM in the world. IGT people can get diseases of cardio-cerebrovascular, microvascular disease, lipid metabolism disorders, metabolic syndrome except DM. This means that diabetic complication has appearanced more early in IGT stage. So, the intervention research of IGT get more and more attention.This project aimed to find if prescription for tonifying Kidney fed during embryonic development can prevent IGT caused by feeding high fat diet, and to provide new ideas of preventing IGT and serious diseases for people all over the world, and to provide easy technology to improve the quality of life and give a good birth and good care. At the same time, this also can provide an evidence of the theory "the kidney being the origin of congenital constitution","kidney controlling reproduction and upgrowth","invigorating the mother to benefit the children","preventive treatment of disease" in Traditional Chinese Medicine (TCM) and lay a solid foundation for making TCM known worldwide.MethodsWistar rats were put in the same cage overnight at the ratio of2:1for female and male. The pregnant rats were randomly divided into5groups. The normal control group and model group were fed by normal saline on the1st day, ig,20mL. kg-1,19d.In other3experiment groups, the pregnant rats were fed respectively with Zuoguiwan (ZGW, a prescription for reinforcing Kidney Yin) or Youguiwan(YGW, a prescription for reinforcing Kidney Yang) or Bazhen decoction (BZD, a prescription for supplementing Qi and blood), ig,20g. kg-1,20mL. kg-119d, at the same time. After birth, one male newborn rat were selected from every brood randomly into new groups and the names of these groups were unchanged. The other four groups except the control group were fed with high fat diet in6th week. 1. After12weeks of high fat diet, the FBG and2hBG were measured. IGT models were successful. All the rats were Killed. The body weight, weight of abdominal adipose tissue, blood lipid were measured to find the effect of high fat diet and prescription for tonifying Kidney and prescription for supplementing Qi and blood during embryonic development on the above measurements.2. FINS, Leptin and APN were measured with double antibody sandwich ELISA to observe the effect of high fat diet and prescription for tonifying Kidney and prescription for supplementing Qi and blood fed during embryonic development on the above measurements.3. IRmRNA, LRmRNA and APNmRNA were measured with fluorescence quantitative PCR to observe the effect of high fat diet and prescription for tonifying Kidney and prescription for supplementing Qi and blood fed during embryonic development on the above measurements.4. Fresh liver tissue of rats were taked, conventional washed, dehydrated, embeded, sliced, HE stained and observed under the light microscopic level. Fresh pancreas tissue of rats were taked and insulin expression in β cell were observed to find the effect of high fat diet and prescription for tonifying Kidney and prescription for supplementing Qi and blood fed during embryonic development on the above measurements.Results1. Effect of high fat diet and prescription for tonifying Kidney and prescription for supplementing Qi and blood during embryonic development on BG, body weight, weight of abdominal adipose tissue and blood lipid.FBG:FBG of model group was significantly higher than FBG of normal control group (P<0.05), but have not yet reached the diabetes diagnosis standard. FBG of BZD group, ZGW group and YGW group was higher than FBG of normal control group too, but no significant difference.2hBG:2hBG of model group was significant ly higher than2hBG of normal control group (P<0.01), the mean>7.8mmol/L, IGT models were successful.2hBG of ZGW group was significant lower than2hBG of model group (P<0.05);2hBG of ZGW group and YGW group was not significantly different from model group. Body Weight:the body weight of model group was significantly higher than the body weight of normal control group (P<0.01), the Body Weight of BZD group, ZGW group and YGW group was higher than normal control group too, but no significant difference.Weight of Abdominal Adipose Tissue:the weight of abdominal adipose tissue of model group was significantly higher than the weight of abdominal adipose tissue of normal control group (P<0.01); the weight of abdominal adipose tissue of ZGW group and YGW group was significant lower than the weight of abdominal adipose tissue of model group (P<0.05); the weight of abdominal adipose tissue of BZD group was not significantly different from model group.Weight of Abdominal Adipose Tissue/Body Weight:weight of abdominal adipose tissue/body weight of model group was significantly higher than weight of abdominal adipose tissue/body weight of normal control group (P<0.05); weight of abdominal adipose tissue/body weight of ZGW group was significantly lower than weight of abdominal adipose tissue/body weight of model group (P<0.05) weight of abdominal adipose tissue/body weight of ZGW group and YGW group was not significantly different from model group.HDL:HDL of model group was significantly lower than HDL of normal control group (P<0.01); HDL of BZD group, ZGW group and YGW group was higher than HDL of model group(P<0.01).LDL:LDL of model group was significantly higher than LDL of normal control group (P<0.01); LDL of BZD group, ZGW group and YGW group was not significantly different from model group.TG:The mean of TG of model group was higher than The mean of TG of normal control group, but was not significantly different from control group.. TG of BZD group,ZGW group and YGW group was not significantly different from model group,but the mean of TG of ZGW group and YGW group was lower than the mean of normal control group, the mean of TG of BZD group was higher than the mean of model group.TC:TC of model group was significantly lower than TC of normal control group (P<0.01); TC of BZD group, ZGW group and YGW group was significantly higher than TC of model group (P<0.01). 2.Effect of high fat diet and prescription for tonifying Kidney and prescript ion for supplementing Qi and blood during embryonic development on FINS, leptin and APN.FINS:FINS of model group was significantly higher than FINS of normal control group (P<0.01); FINS of BZD group, ZGW group and YGW group was s ignif icant ly lower than FINS of model group (P<0.01).ISI:ISI of model group was significantly lower than ISI of normal control group (P<0.01); ISI of BZD group was significantly higher than ISI of normal control group (P<0.05); ISI of ZGW group and YGW group was significantly higher than ISI of model group (P<0.01).HOMA-IR:HOMA-IR of model group was significantly lower than HOMA-IR of normal control group (P<0.01); HOMA-IR of BZD group was s ignif icant ly higher than HOMA-IR of normal control group (P<0.05); HOMA-IR of ZGW group and YGW group was significantly higher than HOMA-IR of model group (P<0.01).Leptin:Leptin of model group was significantly higher than Leptin of normal control group (P<0.01); Leptin of ZGW group and YGW group was significantly lower than Leptin of model group (P<0.01). Leptin of BZD group was not significantly different from model group.APN:APN of model group was significantly lower than APN of normal control group (P<0.01); APN of ZGW group was significantly higher than APN of model group (P<0.01); the APN mean of BZD group and YGW group was higher than model group, but no significant difference.3. Effect of high fat diet and prescription for tonifying Kidney and prescription for supplementing Qi and blood during embryonic development on IRmRNA, LRmRNA and APNmRNA.IRmRNA and LRmRNA:There was no significant difference among IRmRNA and LRmRNA in liver tissue of normal control group and experiment groups. But the IRmRNA and LRmRNA means of model group were lower than the IRmRNA and LRmRNA means of normal control group, the IRmRNA and LRmRNA means of BZD group and ZGW group were higher than the IRmRNA and LRmRNA means of model group.APNmRNA:There was no significant difference among APNmRNA in adipose tissue of normal control group and experiment groups. But the APNmRNA mean of model group was lower than the APNmRNA mean of normal control group, the APNmRNA mean of ZGW group and YGW group was higher than the APNmRNA mean of model group.4. Effect of high fat diet and prescription for tonifying Kidney and prescription for supplementing Qi and blood fed during embryonic development on liver tissue morphology and insulin expression in β cells.Liver tissue morphology: There was no fatty degeneration of liver tissue in normal control group. There was diffused empty bubbles of liver tissue in model group. There was a little empty bubbles of liver tissue in ZGW group. The pathological changes of liver in YGW group and BZD group were between model group and ZGW group.Insulin expression in β cells:The color of positive insulin staining is tan. The shapes of pancreas islets in normal control group were regular and the positive insulin staining lies in the cytoplasm of β cells. The positive β cells arranged regularly in uniform size. The shapes of pancreas islets in model group were irregular in different size. Many pancreas islets were hyperplasia and hypertrophy. The positive β cells arranged irregularly in large size. The shapes of pancreas islets in ZGW group were mainly regular, the shapes of posit ive β cells arranged mainly regularly and there were no obvious hyperplasia and hypertrophy. The pathological changes of pancreas islets in YGW group and BZD group were between model group and ZGW group.Conelusions1. Wistar rats fed high fat diet can get IGT with abnormal blood lip id, insulin resistance,, leptin resistance,lower LNP, fatty degeneration of liver tissue, hyperplasia and hypertrophy of pancreas islets, large β cells and the expression of IRmRNA, LRmRNA and APNmRNA tends to decrease.2. ZGW during embryonic development can prevent IGT caused by feeding high fat diet by wi thstanding the adverse changes of above measurements. These effects may be associated with modifying some genes related to IGT. YGW and BZD during embryonic development can withstand the adverse changes of above measurements partly,but the effects are not as good as ZGW and can not prevent IGT caused by feeding high fat diet. 3. ZGW during embryonic development can prevent IGT caused by feeding high fat diet, which reflect the innate character of IGT--Yin deficiency, and provide an evidence of the theory-"the kidney being the origin of congenital constitution""kidney controlling reproduction and upgrowth","invigorating the mother to benefit the children" in TCM, and reflect the theory "preventive treatment of disease"4. This research provides a new idea of prevent IGT at the viewpoint of TCM. |
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