| Objectives:A type II diabetic mellitus rat model was induced to observe the effects of spleen activating and collaterals regulating decoction on dynamin-related protein 1(Drp-1),serum glutathione(GSH-Px)activity,super oxide dismutase(SOD)activity and malonaldehyde(MDA)content in rat ischiadic nerve,detect the change of Drp-1 gene and protein expression in DM rat ischiadic nerve,discuss the protective role of spleen activating and collaterals regulating decoction on rat ischiadic nerve and the potential mechanism of action,and provide experimental evidence for developing effective drugs for the treatment of diabetic peripheral neuropathy(DPN).Methods:SD rats were selected,injected with streptozocin(STZ)and fed with high fat and high glucose fodder to replicate a DM rat model;these DM rats were randomly divided into model group,positive control group,and high dose,medium dose and low dose group(spleen activating and collaterals regulating decoction).Normal group was also set up.After 16 weeks of intervention,the fasting glucose of each group was detected;high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),total cholesterol(TC)and triglyceride(TG)in serum were determined by completely automatic biochemistry analyzer;serum glutathion peroxidase(GSH-PX)activity of DPN rats was determined by colorimetric method,and serum total SOD activity of DPN rats was determined by xanthine oxidase method.Ischiadic nerve conduction velocity was detected by RT-PCR method;HE staining was employed to observe the pathological change of ischiadic nerve of each group under microscope;Envision immunohistochemical staining method and Western blot was used to detect the expression of Drp-1 protein in ischiadic nerve.Results:In the model group,the glucose level of was obviously increased,the lipid level was abnormal,the TG,LDL-C and TC levels were increased,and the HDL-C level was decreased;after the treatment with spleen activating and collaterals regulating decoction,the TG,LDL-C and TC levels were lowered,and the HDL-C level was increased.The microscopic observation of ischiadic nerve sections showed that the nerve fibers of the normal group were closely arranged,while the nerve fibers of the model group were loosely structured,but the above-mentioned pathological change was alleviated following the treatment with spleen activating and collaterals regulating decoction.The nerve conduction velocity was reduced compared with the normal group,showing a statistically significant difference(P<0.01);following the treatment with spleen activating and collaterals regulating decoction,the nerve conduction velocities of the high,medium and low dose groups were faster than those of controls in the model group,showing statistically significant differences(P<0.05).Compared with the normal group,the expressions of Drp-1 protein and mRNA were up-regulated,serum MDA content was increased,and serum GSH-PX activity and SOD activity were decreased in rat ischiadic nerve tissue of the model group;compared with the model group,the expressions of Drp-1 protein and mRNA were down-regulated,serum MDA content was decreased,and serum GSH-Px activity was increased in rat ischiadic nerve tissue of the three treatment groups.Conclusion:A DPN rat model is successfully replicated.In the model group,serum SOD and GSH-Px activity is decreased,MDA content is elevated,and the expressions of Drp-1 mRNA and protein are up-regulated.This suggests that the possible mechanism by which spleen activating and collaterals regulating decoction prevents and treats DNP may be that SOD and GSH-PX activity is improved to lower the content of MDA and downregulate the expression levels of Drp-1 protein and mRNA.This study confirms that spleen activating and collaterals regulating decoction can improve the pathological state of nerves and repair impaired nerves. |
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