Clinical And Experimental Research Of Tangtongfang Treatment On Diabetic Neuropathy And Vascular Disease

Diabetic peripheral neuropathy and lower extremity vascular disease are two common chronic diabetes complications. Neurological examination revealed that60%-90%of diabetics were accompanied with varying degrees of diabetic neuropathy, and30%-40%of the patients were asymptomatic. Ankle-brachial index examination showed that about19.47%-23.8%of the diabetic patients older than50years were suffered from lower extremity arterial disease. Diabetic vascular disease is the pathological basis of diabetic peripheral neuropathy, it’s also the major cause of the latter’s appearance and aggravation. Moreover, two diseases relate closely, and they are the major factor of diabetic foot and lower limb amputation of diabetic patients, seriously affecting the life quality of patients with diabetes. Recently, Chinese medicine play an important roal in the prevention and treatment of diabetic peripheral neuropathy and lower extremity vascular disease. It can improve the clinical symptoms, increase peripheral nerve conduction velocity and lower limb blood supply, and reduce the incidence of diabetic foot and lower limb amputation. Under the guidance of "three types of dialectical" academic thought, based on the National Natural Science Foundation Project-"to explore the mechanism of traditional Chinese medicine in the prevention and treatment of diabetic peripheral neuropathy from apoptosis mediated by ICAM-1" which is charged by my tutor, this thesis is to observe the treatment efficacy of "Tangtongfang " which is my tutor’s experience prescription, and to explore the treatment mechanism of Tangtongfang on lower extremity neuropathy and peripheral vascular disease. The thesis consists three parts-theoretical research, clinical research and experimental research.1Theoretical researchTheoretical research contains three chapters. Chapter1introduced Professor Lin Lan’s academic thought about diabetes and its complications, the formation of diabetical "three types of dialectical" theoretical system and its clinical applications, the treatment way of diabetic peripheral neuropathy and lower extremity vascular disease of Pro. Lin, and the prescription basis of Chinese medicine "Tangtongfang". Chapter2is about the research progress of diabetic peripheral neuropathy disease these years, including its western pathogenesis, traditional Chinese medicine ("TCM") pathogenesis and TCM treatment method. Chapter3is about the research progress of diabetic lower extremity vascular disease, including its western pathogenesis, TCM pathogenesis and TCM treatment method.2Clinical researchObjective-To observe the clinical efficacy of Tangtongfang treatment on diabetic peripheral neuropathy and vascular disease.Methods-The patients suffering from diabetic peripheral neuropathy (Qi deficiency and blood stasis syndrome) were chosen for observation and then were divided into two groups-treatment group and control group. Besides diet therapy, exercise therapy, we give the patients from the two groups corresponding hypoglycemic, antihypertensive therapy according to their actual situation. The control group was asked to take pancreatic kininogenase enteric-coated tablets240IU three times per day; based on the control group treatment, the treatment group also took Chinese medicine Tangtongfang1dose and twice per day. Both of the groups were treated for3months. Collect the general information of the patients, such as sex, age, height, weight, course of disease, and so on; According to the clinical symptom of TCM questionnaire, every patient was scored before and after treatment; The fasting plasma glucose (FBG),2h postprandial blood glucose (2hPBG), HbAlc, lipids, coagulation, liver and kidney function, the nerve conduction velocity, ankle-brachial index, lower extremity arterial diameter before and after treatment. SPSS18.0is adopted as the statistical software and the significance level is0.05.Results-1.Comparison of scores of TCM clinical symptom-Before treatment, the individual symptom scores of the two groups had no statistically significant difference (P>0.05); after treatment, the treatment group improved more significantly(P<0.05) at the following aspects:fatigued spirit and lack of strength, shortness of breath lazy words, dizziness, numbness of the limbs, limb pain and cold extremities.2.Comparison of blood sugar-Before treatment, FBG、2hPBG、HbAlc of the two groups had no statistically significant difference (P>0.05); after treatment, FBG、2hPBG、HbAlc from the two groups both declined, significantly (P<0.05),but not significantly between the two groups (P>0.05).3.Comparison of blood pressure and BMI-Before treatment, the systolic blood pressure, diastolic blood pressure, body mass index of the two groups had no statistically significant difference (P>0.05); after treatment, the indicators from the two groups both declined significantly (P<0.05),but not significantly between the two groups (P>0.05).4.Comparison of lipids-Before treatment, the lipids of the two groups had no significant difference (P>0.05); after treatment, the lipids from the control group had no significant differences than before (P>0.05); compared with the control group, the CHO、TG、LDL-C of the treatment group declined significantly (P<0.01).5.Comparison of coagulation function-Before treatment, the coagulation of the two groups had no statistically significant difference (P>0.05); after treatment, the indicators from the two groups both improved significantly (P <0.01).Compared with the control group, the thromboplastin time and fibrinogen of the treatment group improved significantly (P<0.01).6.Comparison of nerve conduction velocity-Before treatment, the nerve conduction velocity of the two groups had no statistically significant difference (P>0.05); after treatment, the indicators from the treatment group increased significantly (P<0.01); compared with the control group, the conduction velocity of tibial nerve, peroneal nerve, sural nerve of treatment group increased significantly (P<0.01).7.Comparison of ankle-brachial index-Before treatment, the ankle-brachial index of the two groups had no statistically significant difference (P>0.05); after treatment, the indicators from the two groups both rose significantly (P<0.01); compared with the control group, the treatment group rosed more significantly (P<0.05).8.Comparison of lower extremity vascular diameter-Before treatment, the diameters of femoral artery, popliteal artery, dorsal artery from the two groups had no statistically significant difference (P>0.05); after treatment, the diameters from the two groups both increased significantly (P<0.05); compared with the control group, the treatment group increased more significantly (P<0.05).9. Efficacy comparison-The efficacy of the treatment groups was85.71%, better than the control group(65.91%)(P<0.05).Conclusions-Tangtongfang with pancreatic kallikrein enteric-coated tablets can improve diabetic patients’ clinical symptoms of diabetic neuropathy and vascular disease; can regulate diabetic patients’ lipids level through lowering cholesterol, triglycerides and low-density lipoprotein; can regulate coagulation function through inhibiting of platelet aggregation and lower fibrinogen levels; can lower limb improve blood supply through increasing lower extremity vascular diameter; can improve diabetic patients’ peripheral nerve conduction velocity through increasing the blood supply to the nerve. Compared with control group, we can find that the mechanism of Tangtongfang treatment on diabetic neuropathy and vascular disease has close relationship with lipids and coagulation function regulation, which can reduce lower extremity arterial thrombosis and plaque formation, then improve the patients’ blood supply, then raise diabetic patients’ ankle-brachial index and peripheral nerve conduction velocity, finally improve diabetic patients’clinical symptoms.3Experimental researchObjective-To observe the impact of Tangtongfang on diabetic rats’sciatic nerves and femoral artery after ischemia-reperfusion, and to explore the pathogenesis of diabetic peripheral neuropathy and lower extremity vascular disease, and the protective mechanism of Tangtongfang.Methods-Choose18of110SD rats as normal control group and the left as diabetic group, and the model rats suffering from diabetes were induced by STZ. After feeding for4weeks, choose all the modeling rats suffering from diabetes and keep their abdominal aorta, right iliac artery and the right femoral artery blocked for some time, then take them as the right lower limb ischemia-reperfusion model rats. Ischemia-reperfusion later, the diabetic rats were divided into4groups:model group, Tangtongfang low-dose group, Tangtongfang medium-dose group and Tangtongfang high-dose group. The low, medium and high group will be administered by Tangtongfang with the dose of3.5,7,14times of the adults; the normal control group and model group will be fed by water, once per day for8weeks. Observe the mental status, daily activities, eating, drinking, urine and hair color changes, to weigh the rats once per2weeks, and check the blood sugar from tail once per2weeks. After treatment; detect the level of sICAM-1, MPO, lipids in the rats’plasma, the tail nerve conduction velocity, sciatic nerve conduction velocity and sciatic nerve action potential amplitude of the rats; observe the morphological changes of the sciatic nerve and femoral artery; femoral and sciatic nerve were stained by ICAM-1、 TNF-a、MPO、8-OHdG、tunel respectively after treatment; use Western Blot detection to detect the expression level of ICAM-1and MPO in right sciatic nerve after treatment; use Real Time PCR detection to detect the expression level of ICAM-1mRNA in right sciatic nerve after treatment. SPSS18.0is adopted as the statistical software and the significance level is0.05.Results-1.Comparison of rats’ weight:Before modeled(Week0), there is no significant differences in rats’ weight between the groups (P>0.05); before administered by group(Week5), Tangtongfang groups had no significant difference compared with model group (P>0.05), but lighter than normal group significantly (P<0.01); after administered(Week13), Tangtongfang groups had no significant difference compared with model group (P>0.05), but lighter than normal group significantly (P<0.01).2.Comparison of rats’blood sugar:After modeled, the blood sugar of diabetic rats rose significantly than normal group (P<0.01); before administered by group(Week5), the rats’blood sugar of Tangtongfang groups had no significant difference compared with model group (P>0.05), but higher than normal group significantly (P<0.01); after administered(Week13), the rats’blood sugar of Tangtongfang groups had no significant difference compared with model group (P>0.05), but higher than normal group significantly (P<0.01).3.Comparison of rats’lipids:Compared with the normal control group, the rats’CHO、TG、LDL-C in plasma from model group rose significantly (P<0.01); after Tangtongfang treatment, the rats’CHCK TG^LDL-C from treatment groups declined significantly compared with model group (P<0.05)4.Comparison of expression level of sICAM-1and MPO in plasma:Compared with the normal control group, the level of sICAM-1and MPO from model group rose significantly (P<0.01).After Tangtongfang treatment, the level of sICAM-1and MPO from the treatment groups declined significantly compared with model group (P<0.01).5.Comparison of electrophysiological indicators:Compared with the normal control group, the rats’tail nerve conduction velocity, sciatic nerve conduction velocity and action potential amplitude from model group declined significantly (P<0.01).After Tangtongfang treatment, the rats’tail nerve conduction velocity, sciatic nerve conduction velocity and action potential amplitude from treatment groups rose significantly compared with model group (P<0.01).6. Comparison of morphological observation:Compared with the normal control group, the rats’sciatic and femoral artery from the model group injured significantly. After Tangtongfang treatment, the injury got better significantly.7.Comparison of immunohistochemical staining:Compared with the normal control group, the expression level of ICAM-1、TNF-a in sciatic nerve from model group rose significantly (P<0.01), and the expression level of ICAM-1、TNF-a、8-OHdG in femoral artery rose significantly(P<0.01).After Tangtongfang treatment, the expression level of ICAM-1、TNF-a in sciatic nerve from treatment group declied significantly compared with model group (P<0.05).The expression level of ICAM-1、TNF-a、8-OHdG in femoral artery from treatment group declined, and the difference between Tangtongfang medium-dose group/high-dose group and model group was significant (P<0.05).8.TUNEL Staining:In normal control group, sciatic nerve stained by TUNEL had few apoptotic cells, while there was a large quantity of apoptotic cells in the treatment groups. The difference between the model group and the normal control group was significant (P<0.01).After Tangtongfang treatment, the number of apoptotic cells decreased and the difference between the model group and the treatment groups was significant (P<0.05).9.Western blot detection:Compared with the normal control group, the expression level of ICAM-1and MPO in sciatic nerve from model group rose significantly (P<0.01).After Tangtongfang treatment, the expression level of ICAM-1and MPO in sciatic nerve declined, and the difference between the model group and Tangtongfang medium-dose group/high-dose group was significant (P<0.01).10.Real Time PCR detection:Compared with the normal control group, the expression level of ICAM-1and mRNA in sciatic nerve from model group rose significantly (P<0.01).After Tangtongfang treatment, the expression level of ICAM-1and mRNA in sciatic nerve declined significantly compared with the model group (P<0.01)Conclusions-Limb ischemia and reperfusion can cause peripheral neuropathy and lower extremity vascular disease in diabetic rats simultaneously. The symptom included:morphological changes of the sciatic nerve and femoral artery, sciatic nerve conduction velocity decline and neuronal apoptosis increase. The mechanism is closely related with oxidative stress and enhanced inflammatory response, and the symptom included:the expression level of sICAM-1and MPO in plasma rise, the expression level of ICAM-1, TNF-a and MPO in sciatic nerve rise, the expression level of ICAM-1, TNF-a, and8-OHdG in femoral artery rise. Tangtongfang can ease oxidative stress and inflammatory response of neurovascular tissue through reducing the expression level of sICAM-1and MPO in plasma, the expression level of ICAM-1, TNF-a and MPO in sciatic nerve, the expression level of ICAM-1, TNF-a,8-OHdG in femoral artery. Then the injury in sciatic nerve and femoral artery was eased, and their morphology was improved.