| Purpose:The purpose of this study was to investigate the mechanism of the Modified Guizhi Fuling Pills in the prevention and treatment of painful diabetic peripheral neuropathy,and its effect on HIF-1α and ET-1 levels,so as to provide a deep theoretical basis for the clinical use of traditional Chinese medicine in the prevention and treatment of painful diabetic peripheral neuropathy.Materials and methods:A total of 104 healthy male SD rats with SPF grade and body weight range of 200-220 g were selected.They were adaptive fed for one week before the experiment.During the adaptive period,they were free to eat and drink water,the feeding temperature was kept at 20-25℃,the humidity was kept at 40-50%,the daily average light was 12 h,and the padding was changed on time every day.Twelve rats were randomly selected as blank control group by random number table method.After fasting for 12 h,the other rats were induced into diabetic model by one-time intrapitoneal injection of 2%streptozotocin solution 50mg/kg.The diabetic rats were fed ordinary diet for 4 weeks.PDPN modeling was successful if the mechanical pain threshold or thermal pain threshold decreased by more than 50%.They were randomly divided into model group,pancreatic kininogenase treatment group,the Modified Guizhi Fuling Pills low-dose,medium-dose and high-dose treatment groups.The changes of daily activity state,mental state,blood glucose,body weight,mechanical pain threshold,thermal pain threshold and nerve conduction velocity of rats were observed 0 weeks before administration and 2,4 and 8 weeks after administration.The levels of HIF-1α and ET-1 in serum and sciatic nerve of rats were determined by ELISA,PCR and WB,and statistically analyzed by SPSS software.Results:1.Compared with the normal group,the mental state of rats in the model group was poor,the reaction was slow,the hair color was dark and dull,the body weight increased slowly,and the water intake and urine volume increased significantly;All the treatment groups showed the above symptoms but the degree was less than the model group.2.Blood glucose level in model group was significantly higher than that in normal group(P<0.01);At 4 weeks,blood glucose level in medium and high dose groups was significantly decreased compared with model group(P<0.05);At 8 weeks,blood glucose of rats in the Modified Guizhi Fuling Pills treatment groups decreased significantly compared with model group(P<0.01).3.The body weight of rats in model group was significantly lower than that in normal group after modeling(P<0.01);At 4 weeks,the body weight of the high-dose group was significantly increased compared with that of the model group(P<0.05);at 8 weeks,the body weight of the medium-dose group was significantly increased compared with that of the model group(P<0.05);the body weight of the pancreatic kininogenase group and high-dose group was significantly increased compared with that of the model group(P<0.01).4.The tolerance degree of rats in model group to mechanical stimulation was significantly weakened compared with that in normal group(P<0.01);At 4 weeks,the degree of tolerance to mechanical stimulation in medium and high dose groups was significantly higher than that in model group(P<0.05);At 8 weeks,the tolerance degree of mechanical stimulation in pancreatic kininogenase group and low-dose group was significantly higher than that in model group(P<0.05);The degree of tolerance to mechanical stimulation in medium and high dose groups was significantly higher than that in model group(P<0.01).5.The latency time of foot shrinkage reaction in model group and treatment groups was significantly shorter than that in normal group(P<0.01);At 4 weeks and 8 weeks of treatment,the incubation time of heat contraction foot reaction in pancreatic kininogenase group and the Modified Guizhi Fuling Pills groups was longer than that in model group(P<0.01),and the improvement was most obvious in high dose group(P<0.01).6.The nerve conduction velocity of rats in model group was significantly lower than that in normal group(P<0.01);At 4 weeks and 8 weeks of treatment,MNCV in pancreatic kininogenase group,medium and high dose treatment groups was significantly increased compared with model group(P<0.01).7.After 8 weeks of treatment,the serum ET-1 content in the model group was significantly higher than that in the normal group(P<0.01),and the serum ET-1 content in the pancreatic kininogenase group and the Modified Guizhi Fuling Pills low-dose,medium-dose and high-dose treatment groups was significantly lower than that in the model group(P<0.01).After 8 weeks of treatment,HIF-1αm RNA and ET-1m RNA expression levels in sciatic nerve of rats in model group were significantly increased compared with normal group(P<0.01);The m RNA expression level of HIF-1α in sciatic nerve of the Modified Guizhi Fuling Pills low-dose group decreased,but there was no significant difference compared with model group.The m RNA expression levels of HIF-1 α and ET-1 in sciatic nerve of pancreatic kininogenase group and the Modified Guizhi Fuling Pills medium and high dose groups were significantly decreased compared with model group(P<0.01).After 8 weeks of treatment,the expressions of HIF-1α and ET-1 in sciatic nerve of model group were significantly increased compared with normal group(P<0.01);The expression levels of HIF-1α protein and ET-1protein in sciatic nerve of pancreatic kininogenase group and the Modified Guizhi Fuling Pills were significantly decreased compared with model group(P<0.01).Conclusion:The experimental results showed that the Modified Guizhi Fuling Pills could improve the hyperalgesia of rats with painful diabetic peripheral neuropathy,accelerate the nerve conduction velocity of rats with painful diabetic peripheral neuropathy,and play a certain protective effect on nerve tissue by improving microcirculation. |
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