益气活血化瘀法治疗痛性糖尿病周围神经病疗效机制的研究

【作者】 李长辉；

【导师】 于世家；

【作者基本信息】 辽宁中医药大学 ， 中西医结合临床， 2013， 博士

【摘要】 目的:探讨中药木丹颗粒(糖末宁颗粒剂)对链脲佐菌素(STZ)致痛性糖尿病周围神经病变模型大鼠血清疼痛物质五羟色胺（5-HT）、β内啡肽(β-EP)、组胺、缓激肽的影响；及其对腓肠神中电压门控钠离子通道1.7、1.8、1.9(Nav1.7、Nav1.8、Nav1.9)，辣椒素受体1(TRPV1) mRNA表达的影响。方法:采用链脲佐菌素腹腔注射制作痛性糖尿病周围神经病变大鼠模型,给予木丹颗粒灌胃,以苯妥英钠为阳性对照组,并设正常对照组及模型对照组,观察治疗前及治疗后第3W、5W、9W周痛性糖尿病周围神经病变大鼠血清疼痛物质五羟色胺、β内啡肽(β-EP)、组胺、缓激肽含量的变化；并采用RT-PCR法检测治疗前后对痛性糖尿病周围神经病变大鼠腓肠神经Nav1.7、Nav1.8、Nav1.9、TRPV1mRNA表达的变化。结果：模型组在造模后的3W、5W、9W血清中5-HT、组胺、缓激肽水平明显高于空白对照组，有统计学差异(P<0.01)，而β-EP明显低于空白对照组，有统计学差异(P<0.01)。与模型组比较在3W、5W、9W三个时间点木丹颗粒组和苯妥英钠组血清中组胺、缓激肽水平均明显降低，有统计学差异(P<0.01);血清中5-HT在第3W时木丹颗粒组与模型组比较明显降低，有统计学差异(P<0.05),苯妥英钠组与模型组比较明显降低，有统计学差异(P<0.01)；在第5W、9W木丹颗粒组和苯妥英钠组与模型组比较明显降低，有统计学差异(P<0.01)；3W、5W、9W木丹颗粒组和苯妥英钠组血清中β-EP水平与模型组比较明显呈升高，有统计学差异(P<0.01)。与木丹颗粒组比较：在3W、5W、9W时苯妥英钠组血清中5-HT组明显高于木丹颗粒组，有统计学差异（P<0.05）；组胺水平苯妥英钠组明显高于木丹颗粒组，有统计学差异(P<0.01)；缓激肽水平虽有升高但无统计学差异(P>0.05)；内啡肽水平虽有下降但无统计学差异(P>0.05)。模型组3W、5W、9W时Nav1.7、Nav1.8、Nav1.9、TRPV1的表达呈上升趋势，苯妥英钠组可以下调Nav1.7、Nav1.8、Nav1.9、TRPV1的表达，且在3W、5W、9W呈下降趋势。木丹颗粒组与苯妥英钠组显示出相同的变化趋势。结论：1.本实验在糖尿病大鼠模型基础上成功诱发痛性周围神经病变模型，此模型为今后类似模型提供参考。2.木丹颗粒治疗痛性糖尿病周围神经病变的机制可能与减少血清中致痛物质五羟色胺、组胺、缓激肽的水平的表达，增加止痛物质β-内啡肽的水平表达有关。3.木丹颗粒对痛性糖尿病周围神经病变大鼠电压门控钠离子通道1.7、1.8、1.9通道阻断作用，下调其表达，可能是其改善痛性糖尿病周围神经病症状的机制之一。4.木丹颗粒可减少痛性糖尿病周围神经病变辣椒素受体1（TRPV1）的表达，可能是其改善痛性糖尿病周围神经病症状的机制之一。更多还原

【Abstract】 Objective:To investigate the effect of Mudan granule (Tangmoning granule) on streptozotocin (STZ)induced painful peripheral diabetic neuropathy rats serum pain substance fivehydroxytryptamine (5-HT), β-endorphin (β-EP), effect of histamine, bradykinin; and thevoltage gated sodium sural nerve in ion channel1.7,1.8,1.9(Nav1.7, Nav1.8, Nav1.9),vanilloid receptor1(TRPV1) mRNA expressioMethod：By intraperitoneal injection of streptozotocin model of painful diabetic peripheralneuropathy in rats given orally, Mudan granule, with phenytoin sodium as positive controlgroup, and normal control group and model control group, enkephalin pain rats serum3W,5W,9W weeks of painful diabetic peripheral neuropathy in five., β-were observed before andafter treatment (β-EP), histamine, bradykinin content changes; and using the method ofRT-PCR test before and after the treatment of the sural nerve in Nav rats of painful diabeticperipheral neuropathy in1.7, Nav1.8, Nav1.9, TRPV1mRNA expressioResult：The model group after making the mold3W,5W,9W in serum5-HT, histamine,bradykinin levels were significantly higher than that in the control group, there wasstatistically significant difference (P<0.01), and β-EP were significantly lower than those inthe blank control group, there were significant differences (P<0.01). Compared with themodel group in3W,5W,9W three time points of Mudan granule group and phenytoin serumhistamine, bradykinin levels were significantly reduced, there is significant difference(P<0.01); the serum5-HT in section3W of Mudan granule group compared with the modelgroup decreased significantly, there was statistically significant difference (P<0.05),phenytoin group and compared with model group decreased significantly, there wasstatistically significant difference (P<0.01); in5W,9W Mudan granule group and phenytoingroup and model group decreased, there was statistically significant difference (P<0.01);3W,5W,9W of Mudan granule group and phenytoin serum β-EP level was increased significantlyas compared with the model group, there is statistical differences (P<0.01). Compared withthe Mudan granule group:3W,5W,9W in5-HT group was significantly higher than that ofphenytoin sodium in serum of Mudan granule group, there was statistically significantdifference (P<0.05); histamine level phenytoin group was significantly higher than that ofMudan granule group, there was statistically significant difference (P<0.01); bradykinin levelswere elevated but no statistical difference (P>0.05); inside enkephalin levels were decreasedbut there was no statistical difference (P>0.05Expression of3W of model group,5W,9W Nav1.7, Nav1.8, Nav1.9, TRPV1assumes the trend of escalation, phenytoin group can downregulate the expression of Nav1.7, Nav1.8,Nav1.9, TRPV1,3W,5W, and9W decreased. Mudan granule group and phenytoin groupshowed the same change trend.Knottheory:1the successful model of evoked painful peripheral neuropathy in diabetic rat model, thismodel provides reference for similar model.2mechanisms of Mudan granule in the treatment of painful diabetic peripheral neuropathymay be associated with a decrease in expression of serotonin, histamine, pain in five ofbradykinin induced by serum levels of analgesic substances, increased endorphin levelsexpression.3.Inhibition of3of Mudan Granule on painful diabetic peripheral neuropathy in rats ofvoltage-gated sodium channels1.7,1.8,1.9channel, down-regulating expression, may bethe mechanism of improvement of painful diabetic peripheral neuropathysymptoms of.4of Mudan granule can reduce painful diabetic peripheral neuropathy vanilloid receptor1(TRPV1) expression, may be the mechanism of improving the painful diabetic peripheralneuropathy symptoms of更多还原