Study On The Mechanism Of Action Of Traditional Chinese Medicine Compound Wenluotongsan On Prevention And Treatment Of Peripheral Neurotoxicity Induced By Paclitaxel

Chemotherapy-induced peripheral neuropathy(CIPN)is a serious dose-limiting toxicity of many Anti-neoplastic agents.It severely impairs cancer survivors'quality of life.So far there are no agents proved effective for the prevention or management of CIPN.And the most effective management of CIPN is still dose deduction or regimen suspending.The reason of this situation is that the mechanisms of CIPN are still not explicit.Therefore,it's very important and urgent to understand the mechanisms of CIPN,and speeding the process of discovering for eutherapeutic drug or treatment.The decoction of this study was Wen-Luo-Tong(WLT),which was a clinical empirical formula of Oncology Department of China-Japan Friendship Hospital for CIPN.Previous RCT study,which enrolled 102 CIPN patients,demonstrated that WLT could reduce the pain of peripheral neurotoxicity induced by chemotherapy(pain relief rate:85.07%),improved the quality of life,and with good safety.Other experimental study showed that WLT had neuroprotective effects that improved OXA induced behavioral changes as well as preventing the decrease of NGF level,SNCV,and neuron cell aptosis without affecting the anti-tumor effects of oxaliplatin.On the basis of preliminary clinical research and animal experiment,this study explores the mechanisms of WLT alleviating CIPN.Objectives:1.To verify the therapeutic effects of WLT on peripheral neurotoxicity induced by paclitaxel.2.To clarify the material basis for the treatment of the peripheral neurotoxicity of paclitaxel.3.Explore the mechanisms,predict targets,and provide clues for further research.Methods:The rat model of the peripheral neurotoxicity of paclitaxel was established following the methods from literatures.Animals of PTX were intraperitoneally(i.p.)injected(8 mg/kg,cumulative dose of 24mg/kg)on 3 alternate days(dl,4,7).Animals of WLT and Control received an equivalent volume of saline.Animals took pediluvium of WLT or water for 30min,twice a day.The intervention was initiated 1 day before paclitaxel administration and lasted for 11 days(d0-10).Mechanical paw withdrawal threshold(PWT)was tested before,during,and after paclitaxel treatment(d0,2,4,6,8,10)by an experimenter blinded to treatment groups.Blood and tissue samples were collected on day 10.Blood samples were drawn from the abdominal aorta,and L4-6 spinal cord segments were removed.Sections of the L4-6 spinal cord were processed for immunohistochemistry using CX3CL1.Results of pathological IOD scores were analyzed using IPP softwarre.UPLC-ESI-MS was used to analyze blood samples.According to significant metabolic disturbance of PIPN and retrimed alteration by WLT,the material basis of the pharmacodynamics of WLT was analyzed.Further,with the help of bioinformatics tools,metabolite-protein interaction networks were constructed.The mechanisms of WLT on CIPN management were explored.Results:1.According to the results of behavioral tests,immunohistochemistry,and UPLC-ESI-MS analysis,the animal model of paclitaxel induced peripheral neuropathy was successfully duplicated.2.Administration with paclitaxel had induced marked mechanical allodynia on PTX and WLT since day 2.Statistical difference between PTX and Control had been shown since day 4,while that of between PTX and WLT had been shown since day 6.(P<0.05)It demonstrated that WLT attenuated the PWT decrease of paclitaxel-induced peripheral neuropathy.3.Immunochemistry staining showed that paclitaxel increased the expression of CX3CL1 in neurons and dendrites of spinal cord sections.Meanwhile,compared with PTX group,WLT markedly down-regulated the expression of CX3CL1.(P<0.01)4.UHPLC-ESI/MS analysis identified the Metabolic Disturbances of Paclitaxel induced Peripherial Neuropathy,and marked 19 different metabolites between PTX and Control.5.The linoleic acid and glycerophospholipid pathways were identified as targets of paclitaxel induced peripheral neuropathy.WLT attenuated paclitaxel induced peripheral neuropathy by regulating these two pathways,specifically down-regualting linoleic acid and up-regulating PC(20:4(5Z,8Z,11Z,14Z)/18:0),LysoPC(20:5(5Z,8Z,11Z,14Z,17Z)).Conclusions:1.WLT attenuated paclitaxel induced peripheral neuropathy by alleviating the pain and down-regulating the expression of CX3CL1 in spinal cord.2.The linoleic acid and glycerophospholipid pathways were related to paclitaxel induced peripheral neuropathy.Linoleic acid might be risk factors of PIPN.3.The linoleic acid and glycerophospholipid pathways were identified as targets of paclitaxel induced peripheral neuropathy.