10281064 2005 5
1. 2. 1. SD STZ60mg/kg 3 >11.1mmol/L 7 0 3 2.25g/kg d 19 2. SD 10 20 5 10 10, 2.25g/kg d 22 : 2:1 21 23 1/3 1/4 22 INS MDA 21 CAs 45
60 W256 2 IgG IgM TNF- 1. 2.25g/kg d 2. 2.25g/kg d DA NE >30% 1. 2. 3. 1
Preliminary Study On Eugenicpharmacological Effects of Liuwei Dihuang Pill Postgraduate Wei, Han Advisor Associate Professor Jiang, Wenyue Department Department of Integrated Traditional Chinese & Western Medicine Abstract Aim To study the effects of Liuwei Dihuang Pill (LDP) on the offsprings of gestational hyperglycemia and hot constitution rats. Contents 1. To study the effects of LDP on pregnant rats with severe hyperglycemia and their offsprings. 2. To study the effects of LDP on hot constitutions rats and their offsprings. Methods 1. To investigate effects of LDP on pregnant rats with severe hyperglycemia and offspring. SD female rats were induced hyperglycemia by an i.p. injection of STZ (60mg/kg), and fasting PG levels were tested 3 days later, PG>11.1mmol/L were chosen as model, 1w after the injection, the rats were mated with the male. The morning that conception was verified by the presence of sperm in a vaginal smear was designated gestational day 0. On gestational day 19, maternal blood was taken from abdominal aorta and fetuses were separated by c-section and observed. On gestational day 3, half of models were treated with 2.25g/kg d LDP. 2. To study the effects of LDP on hot constitutions rats and offspring. According to the anus temperature in 6 consequent days female rats were separated into two groups: cold and hot constitution. The male were also distinguish as the same way: lower and higher temperature. Fasting PG was tested among females. In the following 22 days, 10 hot female rats were medicated with 2.25g/kg d LDP, other 2
female rats with normal feeds as control group. Then anus temperature, fasting PG and urine catecholamines(cas) was determined. Two female mated with one corresponding male. Parturition progress was monitored and then each of the pup was weighed. 1/3 1/4 newborn rats from each dam were killed at 22h after delivery, and their PG, INS, MDA levels were analyzed. 21days after delivery part of the other pups PG and urine CAS were measured. Some other pups anus temperature, PG and INS levels were measured 45 days after weaning. Walker 256 (W256) carcinosarcoma cells were implanted subcutaneously 60 days after delivery. Two weeks later, rats were killed to test IgG, IgM, TNF-α and weigh Spleen, Thymus. Results 1. Compared with control group, the plasma glucose, body weight, body length, tail length, and tail length in fetuses from dams with severe hyperglycemia decreased markedly, while the insulin content in pancreas and placenta weight increased. LDP at the dose of 2.25g/kg d could modulate the above parameters to varying degrees. 2. Compared with hot constitution control, LDP at dose of 2.25g/kg d could increase the weight of hot constitution newborn rats. With aging, the weight of pups from the medicated dams was lower than that of offspring from hot constitution control, who showed catch-up tendency; significantly decreased PG in newborn rats, but with aging, statistic difference disappeared; significantly decreased INS in newborn rats, and had no statistic difference with the cold constitution ones, at some degree increased INS of hot constitution pups; significantly decreased DA and NE of pups, had no statistic difference with the cold constitution ones, and decreased E of pups; decreased temperature to some degree; significantly decreased MDA in newborn rats; decreased carcinoma weight to some degree, the rates of anti-carcinoma were above of 30%. Conclusions 1. LDP could modulate the decreased level of weight, body length, tail length, PG and increased level of INS, placenta weight in fetus led by severe maternal hyperglycemia. 2. LDP could modulate the decreased level of birth weight, INS and increased level of PG, MDA, temperature and urine catecholamines in F1 led by hot constitution. 3. LDP could inhibit the F1 carcinosarcoma growth in maternal hot constitution. Key words LDP; maternal hot constitution and cold constitution; maternal hyperglycemia; blood glucose; offspring 3
1 6 9 11 16 32 37 40 60 62 63 4
2000 60% 2020 3/4 28 2 40 1 Barker 2 3,4.. 5 6,7 5
8 9 - - HPA HPA 10 11 12 2 13 2 1958 14 15,16 17 18,19,20,21,22 23 6
( ), 24 25,26 27,28 29,30 31 29 32 32 33 2 34,35,36,37 7
8 1. SD 190 10g SYXK( )2002-0002 22 2 50 10 2. 0309134 STZ Sigma PH 4.4 0.1M 20010306 20000312 W256 860901 HCL 20030928 20030806 NAOH 20021018 EDTA 20030306 20030568 20020506 030427 20030920 20030780
E Sigma 2 8 DA Sigma NE H31021177 750ml 250ml 15ml ph5.4 EDTA3.74g 1mol/L 100ml ph8.2 5mol/L NAOH ph 8.2 250mg 5g 100ml 25 55mol/L NAOH 1:9 38 Anton 39 100g 1000ml 2mol/L HCL500ml 90 45min 2mol/L HCL250ml 70 10min 2mol/L HCL500ml 50 45min 200ml/ ph 3.4 120 1h 200 2h 3. 4. MC 3 2 CARY Eclipse LG-R-80 ZK380 BHG LD4 2 BP211D 0.1mg ph CHM86801 9
1. 1 SD 60 54 STZ60mg/kg 12 3 >11.1mmol/L 40 2 STZ 7 17:00 8:00 0 42,41 3 3 24 /60kg d 15 8 3g 2.25g/kg d SD i.p. 7 i.p.stz60mg/kg CT PG>11.1mmol/L 3 0 19 PG,ING,LP, PG,ING 1 LDP Fig 1 Flow chart of studying effects of LDP on pregnant rats with severe hyperglycemia 10
4 19 2 400mg/kg 1 2. 1 150 SD 6 x x -1 x +1 42 10 20 2 30 5 5 10 SD 2.25g/kg d 22 2 LDP Fig 2 Flow chart of studying effects of LDP on F0 rats of hot constitution 11
(3) 22 1 3 4 : 2:1 5 8:00 0 5 21 23 43 21 45 60 2 W256 IgG IgM TNF- 3 LDP Fig 3 Flow chart of studying effects of LDP on F1 rats of hot constitution 6 1/3 1/4 22 21 45 5 60 12
2 IgG IgM TNF- 7 44 W256 Wistar 1 2 10 7 0.2ml/ 60min IGG IGM TNF-, SI TI 3. 1 9 11 45 2 CAs E NE DA CAs CAs CAs 24 CAs 46 47 24 6mol/LHCL PH 3 1 0.5gEDTA 5mol/L NAOH PH 8.5 1g 15 2000rpm 5 0.1mol/L 4ml 10 1ml 0.25mol/L1ml EDTA0.2ml PH5.4 3.5 Y 1.2ml PH8.0 6.0 0.2ml 2.5 0.2ml 2.5 5mol/L 0.5ml 2 13
A 420 497nm NE 395 475nm.NE 10, 330 374nm DA E NE300ng DA500ng 0.01mol/LHCL 200 g/ml 48,49 3 14 2 3000r/min 15min 4 INS 50 6ml 4 600r/min 30min 3000r/min 15min 5 MDA, 4 10% TBA 51 6 IgG 3000r/min 15min 7 IgM 3000r/min 15min 8 TNF- 3000r/min 15min 9 a10 a150 10S -1 150S -1 1ml 10-20IU/ml 4. SPSS11.5, T Person 14