| Diabetic nephropathy (DN) is the most common diabetic microvascular complications. About 20% ~ 40% people with DM will develop to DN, that is one of the leading death causes of patients. Urine protein increasing and glomerular sclerosis are the main features of DN.Microcirculation is the blood circulation between arteriole and veinule. Microcirculation disturbance is the abnormality of shape and function of blood vessel and blood flow velocity. Microcirculation disturbance is closely associated with the initiation and development of diabetic nephropathy.The microcirculation disturbance of DN patients will result in thromboxan increasing, hyperfunction and aggregation of paltelet, vasoconstriction, hypercoagulability and slowing of the blood flow, which will make the blood flow standstill, and the oxygen, nutrient and energy can not be transported effectively. All of these will lead to aggravation of the DN, and the end-stage renal disease occuring.Shuangqing is made up of seven kinds of traditional Chinese medicines. They are semen cassiae; solomonseal rhizome; mulberry leaf; chrysanthemi flos; pueraria; batatatis rhizome; phragmitis rhizome. Many of them have remarkable effect on activating blood circulation and dissipate blood stasis. They can soften blood vessel, improve microcirculation, prevent and cure microcirculation disturbance.Our research is to observe the effects of shuangqing on diabetic nephropathy in rats and the microcirculation of the ala auris of mice.Part 1 Effect of shuangqing on diabetic nephropathy in ratsObjective: To observe the preventive and therapeutic effect of shuangqing on the FPG increasing and kidney injury of early diabetic nephropathy rats. Methods: 80 Healthy male Wistar rats (220±20g) were selected. Eleven rats were left as normal control group, and the others were made diabetic nephropathy model. After anaesthetizing the rats with 1% pentobarbital, we removed their left kidney, and then gave them 1ml benzylpenicillin sodium to protect them against infections. Two weeks later, without food for 12 hours, rats were treated by intraperitoneal injection with streptozotocin (STZ)- citric acid balance solution 50mg/kg while the rats in the normal control group were treated by intraperitoneal injection with citric acid balance solution by the same volume. Blood samples were obtained from fossa orbitalis vein of rats and fasting plasma glucose (FPG) was determined three days after STZ was given. Rats whose blood glucose was higher than 11.1mmol/L were selected and divided into 5 groups according to FPG and body weight: (1) Positive control group (valsartan 8mg/kg + glibenclamide 1mg/kg); (2) High dose of Shuangqing group(SQ-H, 4g/kg); (3) Middle dose of Shuangqing group (SQ-M, 2g/kg); (4) Low dose of Shuangqing group (SQ-L, 1g/kg); (5) Model group (AD). The rats in normal group were given distilled water. All of the rats were treated by intragastric administration with 1ml/100g everyday for 4 weeks.Results:1 The condition and body weight of the rats: The rats in model group were of polydipsia, polyphagia, polyuria and emaciation. The symptom of"three mores and one less"was obvious. The rats were depressed and slow in reacting, and their fur was dull. The rats in normal group were in good condition. The symptoms of the rats in positive and shuangqing groups were not so serious as that in model group.At the beginning of the experiment, body weight of the rats in each group was similar. But 4 weeks later, the body weight of the rats in normal (321±23) group was higher than that in model group (225±37, P<0.05). The body weight of the rats in positive (298±49) and SQ-H (279±46) groups was higher than that of the rats in model group (P<0.05).2 The change of FPG: After 4 weeks, the FPG in model group (35.2±10.9) was higher than that in normal group (4.3±0.7) (P<0.05). The FPG in SQ-H, SQ-M, SQ-L groups and positive group was 16.3±5.4, 17.0±9.6, 19.2±7.8 and 6.5±2.2, respectively, all of them were lower than that in model group (P<0.05).3 The indexes of kidney: After 4 weeks, the ratio of kidney weight to body weight (KW/BW) of the rats in model group (0.64±0.07) was higher than that in normal group (0.25±0.04, P<0.05). The KW/BW in positive group (0.46±0.09) and SQ-H (0.51±0.06) group was lower than that in model group (P<0.05). The content of 24h urine protein (Pro) of rats in model group (227±47) was higher than that in normal (32±12) group, the urine Pro of rats in positive (31±12), SQ-H (120±47) and SQ-M group(83±63) was lower than that in model group (P<0.05). BUN in serum of rats in model group (12.0±2.1) was higher than that in normal group (6.0±2.1, P<0.05), and it was higher than that in positive group (7.7±1.4, P<0.05).4 The pathological section: In model group, the glomerulus became bigger, the edema of the renal tubular epithelial cells and the hyperemia of the renal interstitium were serious. Compared with the pathological section of model group, the glomerulus in positive group was smaller, the edema of the renal tubular epithelial cells and the hyperemia of the renal interstitium were lighter. The normal group was regular. The indexes in SQ-H and SQ-M groups were better than that in model groups. SQ-L had no significant effect on the pathological change.Conclusion: Shuangqing can decrease the blood glucose of the early diabetic nephropathy rats. It can protect the kidney of the rats in a dose-dependent manner. Part 2 Effect of shuangqing on the microcirculation of the ala auris of miceObjective: Observe the effect of shuangqing on microcirculation.Methods: Kunming mice (18~22g) were divided randomly into 5 groups (n=8): Normal group (normal saline, NS); Positive group (50mg/kg of Compound Danshen Tablet, CDT); SQ-H (6.4g/kg); SQ-M (3.2g/kg); SQ-L (1.6g/kg). Drugs were given by intraperitoneal injection with 0.2ml/10g. The ala auris of the mice was depilated by adhesive plaster after the mice were anaesthetized by 1% pentobarbital. The mice were fixed on the observation desk and some paraffin was given onto the ala auris. The changes of the speed of blood flow and the diameter of arteriole and veinule were observed after drugs were given. Photos were taken at 0, 10, 20, 30min by microcirculation microscope, and then the diameter of blood vessels was measured.Results:1 The diameter changes of veinule (V): NS had no significant effect on diameter of veinule. The diameter in positive group at 10 min, 20 min and 30 min after drug administration was 50.81 (P<0.05), 50.69 (P<0.01) and 50.44 (P<0.01), respectively, which was bigger than that at 0 min (48.44). SQ-L had no significant effect on diameter of veinule. The diameter in SQ-M group at 10 min (53.61), 20 min (54.00) and 30 min (53.44) after drug administration was bigger than that at 0 min(50.50) (P<0.05). The diameter in SQ-H group at 10 min (59.00), 20 min (59.21) after drug administration was bigger than that at 0 min (56.36) (P<0.05), but the diameter at 30 min had no significant difference with 0 min.2 The diameter changes of arteriole (A): NS had no significant effect on diameter of arteriole. The diameter in positive group at 10 min, 20 min and 30 min after drug administration was 30.00 (P<0.05), 31.50 (P<0.01) and 32.00 (P<0.01), respectively, which was bigger than that at 0 min (27.69). SQ-L had no significant effect on diameter of arteriole. The diameter in SQ-M group at 10 min, 20 min and 30 min after drug administration was 24.67 (P<0.05), 24.94 (P<0.05) and 25.89 (P<0.01), respectively, which was bigger than that at 0 min (21.11). The diameter in SQ-H group at 20 min (35.50), 30 min (35.57) after drug administration was bigger than that at 0 min (32.50) (P<0.05), but the diameter at 10 min had no significant difference with 0 min.3 The changes of blood flow: In NS and SQ-L groups, the numbers of blood capillary didn't increase obviously after drug administration. But the blood flow became faster and more blood capillaries were found after drug was given in SQ-H, SQ-M and positive control groups.Conclusion: Shuangqing can dilate the blood vessels of the ala auris of mice in a dose-dependent manner.
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