| Purpose: Observe the sugar diabetes peripheral neuropathy of model rats nerve protection;In each rat dorsal root ganglion of key protein expression level of BDNF/Trk B signaling system testing,explore Yi kang sugar diabetes peripheral neuropathy of intervention of BDNF/Trk B signaling system in rats;Yi kang sugar regulation of BDNF/Trk B signaling pathways downstream of ERK/CREB regulatory role;Yi kang sugar regulation of BDNF/Trk B signaling system another signaling pathways downstream of PI3K/Akt regulation function.Material and method:Adaptive feed rats after a week,will be 60 rats were randomly divided into two groups: the blank control group(8),module(52).Using high fat feed joint method of intraperitoneal injection of STZ induced diabetic rats model,the model can successfully copy(copy success only 41),after the module is to be built again,rats were randomly divided into four groups: model control group(11),high dose group(10),middle dose group(10),low dose group(10).According to the literature method made mould in diabetic rats.Building modules were given a high-fat feed 52 rats,4 weeks after continuous feeding,fast earlier in the day in STZ injection water,the next morning the abdominal cavity injection chain urea with cephalosporins(STZ,according to the 45 mg/kg),72 hours after detecting fasting glucose injection.Blood glucose control is at target,continue to high fat feed.Injection of STZ after 72 hours test module rats fasting glucose,fasting blood sugar above 16.7 tendency/l as diabetic rats model in rats.Blank control group: do not make processing,feed conventional normal feeding until the end of the experiment.Model control group: do not make processing,high fat feed until the end of the experiment.: the high dose group with high dose of Chinese medicine medicinal broth to fill the stomach through the mouth,2 ml / 100 g weight,once per day,high-fat feed until the end of the experiment.Dose group: to the dose of Chinese medicine medicinal broth to fill the stomach through the mouth,2 ml / 100 g weight,once per day,high-fat feed until the end of the experiment.Low dose group,with a low dose of Chinese medicine medicinal broth to fill the stomach through the mouth,2 ml / 100 g weight,once per day,high-fat feed until the end of the experiment.After the last delivery,hot plate method after the pain threshold,with 10% chloral hydrate(0.3 ml / 100 g)intraperitoneal injection of anesthesia,the determination of nerve conduction velocity,put to death in the rat,bilateral lumbar dorsal root ganglion,side soaked in 4% paraformaldehyde solution in 4 keep spare;On the other side in 2 ml of cryopreserved tubes,cryopreserved in ?-80 refrigerator,set aside.Using glucose meter measuring each rat peripheral blood sugar ?levels;Hot plate method is used to detect each group big mouse hot plate response latency;Adopting biological signal acquisition system was developed for the determination of each rat nerve conduction velocity;Using TUNEL method to detect each cell apoptosis in rat dorsal root ganglion;By using enzyme-linked immunosorbent determination of each group rats serum BDNF levels.Adopt western blot method to detect each group of BDNF in rat dorsal root ganglion,Trk B,p-Trk B,ERK1/2,p-ERK1/2,CREB,p-CREB,PI3 K,and Akt,p-Akt expression level.Results: 1.The model evaluation results: blank control group rats blood sugar,hot plate and build module response latency and nerve conduction velocity detection results show that the built module rats a significant rise in blood sugar levels,hot plate reaction significantly prolong the incubation period,slow nerve conduction velocity(p < 0.01).2.After drug intervention,the big mouse hot plate test results show that compared with blank control group,the rest of the group big mouse hot plate reaction significantly prolong the incubation period,the differences were statistically significant(p < 0.01);Compared with model control group,high,medium and low doses of yi kang intervention group rats sugar hot plate response latency was significantly shortened,statistically significant(p < 0.01);Comparison between different dose groups,have a statistically significant difference(p < 0.01),hot plate response latency shortened with yi kang increased doses of sugar.3.Each group rats nerve conduction velocity detection results show that compared with blank control group,the rest of the group rats nerve conduction velocity was significantly slowed,statistically significant(p < 0.01);Compared with model control group,high,medium and low doses of yi sugar health intervention group rats nerve conduction velocity significantly faster,with statistical difference(p < 0.01);Compared with low dose group,high dose group of nerve conduction velocity of rats increased,with difference(p < 0.05).4.Each cell apoptosis in the dorsal root ganglion of the detection results show that the blank control group accidental apoptosis in rat dorsal root ganglion cells,model control group is a large number of apoptotic cells in rat dorsal root ganglion.Compared with blank control group,the rest of the group in the dorsal root ganglia of rats and the increasing number of apoptotic cells,with significant difference(p < 0.01);Compared with model control group,the treatment group of apoptotic cells was significantly reduced,with significant difference(p < 0.01);Three dose treatment groups apoptotic cells was negative correlation with dose,random quantity increase of apoptosis cells was significantly reduced(p < 0.01).5.Each group of rats serum BDNF levels determination results showed that compared with blank control group,the rest of the group in the rat serum BDNF levels significantly reduced,statistically significant(p < 0.01);Compared with model control group,medium dose and high dose group of rats significantly increased serum BDNF levels,statistically significant(p < 0.01);Compared with low dose group,medium dose and high dose group of rats significantly increased serum BDNF levels,statistically significant(p < 0.01 or 0.05);Compared with dose group,high dose group of BDNF levels increased,with difference(p < 0.05).6.Each rat dorsal root ganglion the expression level of BDNF,Trk B and p-Trk B test results shows that: compared with blank control group,model control group and three doses of the treatment group rats the expression level of BDNF in the dorsal root ganglia,Trk B significantly lower(p < 0.01),Trk B phosphorylation level also significantly lower(p < 0.01);Three dose treatment group compared with model control group,BDNF,Trk B and p-Trk B expression level rise significantly(p < 0.01 or 0.05);Compared with low dose group,medium dose group Trk B expression level rise significantly,high dose group of BDNF,Trk B and p-Trk B expression levels were significantly higher(p < 0.01);Compared with dose group,high dose group,the expression level of BDNF raised(p < 0.05).7.Immunofluorescence double standard method to detect the dorsal root ganglion of BDNF and Trk B expression level the result shows: the blank control group microscopically amounts of BDNF,Trk B,and double positive expression cells,through the microscopic count number of double positive cells,compared with blank control group,model control group and three dose treatment groups of BDNF in rat dorsal root ganglion,Trk B double positive cells was significantly reduced(p < 0.01);Compared with model control group,middle,high dose treatment group of BDNF,Trk B double positive cells was significantly increased(p < 0.01);Low dose group,high dose treatment group of BDNF,Trk B double positive cells was increased(p < 0.01 or 0.05).8.The dorsal root ganglion ERK1/2 expression level test results shows that: compared with blank control group,model control group,low dose group,middle dose group ERK1/2 expression in dorsal root ganglia of rats were significantly reduced(p < 0.01);Compared with model control group,low,medium and high dose group of ERK1/2 expression level are different degree rise,statistically significant(p < 0.01);With the increase of dose,ERK1/2 expression level was also significantly increased,and the comparison between groups,statistically difference(p < 0.01 or 0.05).9.Dorsal root ganglion ERK1/2 expression level test results shows that: compared with blank control group,model control group,low,middle dose group of ERK1/2 expression in dorsal root ganglia of rats were significantly reduced(p < 0.01);Compared with model control group,low,medium and high dose group of ERK1/2 expression level are different degree rise,statistically significant(p < 0.01);With the increase of dose,ERK1/2 expression level is raised,the comparison between groups,statistically significant(p < 0.01 or 0.01).Dorsal root ganglia p-ERK1/2 expression level test results shows that: compared with blank control group,model control group,low dose group rats p-ERK1/2 expression in dorsal root ganglia were significantly reduced(p < 0.01);Compared with model control group,high dose group,medium and low p-ERK1/2 expression level are different degree rise,statistically significant(p < 0.01);With the increase of dose,p-ERK1/2 expression level increased significantly,also is compared between each group,there was statistical difference(p < 0.01).10.The dorsal root ganglion after CREB,p-CREB expression level test results shows that: compared with blank control group,model control group,low dose group,middle dose group after CREB expression in dorsal root ganglia of rats were significantly reduced(p < 0.01);Compared with model control group,high dose group,medium and low after CREB higher expression level are different degree,there are statistically significant(p < 0.01);Significantly increased with the increase of dose,p-CREB expression level also,comparison between each group,with statistical difference(p < 0.01 or 0.01).Compared with blank control group,model control group,low dose group p-CREB expression in dorsal root ganglia of rats were significantly reduced(p < 0.01),high dose group of p-CREB expression level was significantly increased(p < 0.05);Compared with model control group,high dose group,medium and low p-CREB higher expression level are different degree,there are statistically significant(p < 0.01);With the increase of dose,p-CREB expression level was also significantly increased,and the comparison between groups,with difference(p < 0.01 or 0.05).11.Dorsal root ganglion PI3 K expression level test results shows that: compared with blank control group,model control group,low dose group,middle dose group of PI3 K expression in dorsal root ganglia of rats were significantly reduced(p < 0.01 or 0.05);Compared with model control group,PI3 K expression level low and middle dose group were different degree rise,statistically significant(p < 0.01);Significantly increased with the increase of dose,PI3 K expression level also,comparison between each group,with difference(p < 0.01 or 0.05).12.The dorsal root ganglion Akt,p-Akt expression level detection results show that compared with blank control group,model control group,low dose group,middle dose group Akt expression in dorsal root ganglia of rats were significantly reduced(p < 0.01 or 0.05);Compared with model control group,low Akt expression level in middle and high dose group were different degree rise,statistically significant(p < 0.01);Significantly increased with the increase of dose,Akt expression level also,comparison between each group,with difference(p < 0.01 or 0.05).Conclusion: 1.Renal sugar kang can shorten diabetes peripheral neuropathy model rats hot plate response latency,increase the sciatic nerve conduction velocity.2.Renal sugar kang can reduce diabetes peripheral neuropathy model rat dorsal root ganglion cell apoptosis in quantity.3.Kidney,can increase the sugar diabetes peripheral neuropathy model rats serum BDNF levels.4.Diabetes peripheral neuropathy complications and the expression level of BDNF/Trk B in dorsal root ganglia are closely related.5.Renal sugar kang could be by adjusting the DPN model rat dorsal root ganglion of the expression of BDNF/Trk B achieve its antiapoptotic effect,but the specific regulation pathway is unclear.6.In the process of diabetic peripheral neuropathy,ERK/after CREB signaling pathway is restrained,the change may be related to increased number of apoptosis in the dorsal root ganglion,speculated that the mechanism may be due to after CREB phosphorylation of regulating the expression of BDNF levels.7.Yi kang granule for sugar regulation effects of ERK/after CREB signal pathway,the mechanism may be related to yi sugar kang granule has the dorsal root ganglion cells apoptosis related,speculated that the regulation may be due to the yi kang can activate the ERK/sugar after CREB signaling pathways,multiply after CREB phosphorylation,regulate the expression of BDNF levels rise,thus play a role of dorsal root ganglion cell apoptosis.8.Diabetes peripheral neuropathy of PI3 K signaling pathway may be suppressed,thus resulting in a loss of key protein after CREB phosphorylation level downstream,reduced the expression of BDNF,cause the dorsal root ganglion cell apoptosis.9.Yi sugar of PI3 K signaling pathways may have activation effect,at the same time promote the downstream key protein after CREB phosphorylation level rise,increased the expression of BDNF,play a role of resistance to the dorsal root ganglion cell apoptosis. |
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