The Clinical Efficacy And Mechanism Of Kidney Tonification And Detoxification Party On The Treatment Of Multiple Myeloma And Experimental Study

ObjectiveMultiple myeloma is a primary malignant tumors of hematopoietic system,although the overall survival rate has improved,but patients with the bone pain and renal failure still affect the no event survival.At present,the main clinical method is still the chemotherapy for the treatment of multiple myeloma.But multi-resistant phenomena appeared in the process of chemotherapy,the patients with remission rate and disease-free survival rate drops greatly,and end up with a relapse.with the introduction of new drugs and hematopoietic stem cell transplantation,which constructed the "induction,consolidation,maintenance" treatment,the curative effect of patients had the very big enhancement,but so far,the multiple myeloma is still an incurable disease.Multi-resistant phenomena is the most important factors of recurrent multiple myeloma.Use of traditional Chinese medicine,therefore,reduce the side effects of chemotherapy,enhance chemotherapy for efficiency,improve the quality of life,improve survival without events,is one of the best ways to improve the curative effect of multiple myeloma,therefore,research of attenuated synergies drugs and methods in the research field of multiple myeloma attract widely attention.Multiple myeloma belong to the category of "involving the bone" of traditional Chinese medicine theory,its pathogenesis is " kidney d eficiency and heat toxin".Therefore,we according to the clinical practice for many years,formed the kidney tonification and detoxification,composition is:banzhilian 15 g,spreading hedyotis herb 30 g,shancigu 15 g,shudi 20 g,prepared rhizome of rehmannia,arrowhead,fresh mushroom mountain yam 30 g,mountain cornus 15 g,radix dipsaci 15 g,20 g drynaria fortunei,malaytea scurfpea fruit 15 g,rhizoma zedoariae,15 g.We found that the party has a certain clinical curative effect.So,this research intends to further clarify the kidney tonification and detoxification clinical curative effect and mechanism of action,create a new path for traditional Chinese medicine treatment of multiple myeloma.Based on the above theory,this study employs the kidney tonification and detoxification treatment in patients with multiple myeloma,from genetics,fluid cytology,cellular immunological indexes to stratification of patients,the clinical curative effect of evaluation,and observe the kidney detoxification effect on myeloma cells of molecular biological indicator,illustrate the kidney detoxification efficacy in patients with multiple myeloma,and tries to illustrate the kidney detoxification of molecular targets.Including clinical and experimental research,this study aims to observe the kidney tonification and detoxification cooperation to improve effect and decrease toxicity of chemotherapy in the treatment of multiple myeloma,and to explore its molecular mechanism,the efficacy of multiple myeloma multiple myeloma cell lines on the preliminary research on kidney detoxification party RPMI8226 cell in inhibiting growth and inducing apoptosis of the role of,and access to treatment for kidney tonification and detoxification beneficial cytology,molecular biology basis for multiple myeloma.MethodsThis study is divided into two parts:clinical and experimental research:1 clinical research:Kidney tonification and detoxification party to study the clinical effect of the treatment of multiple myelomaMethodsinto the group between January 2015 and December 2015,the first affiliated hospital of guangzhou university of TCM haematology department in hospital and outpatients,in line with the diagnostic criteria of multiple myeloma was 60 cases,in accordance with the principle of random divided into the experimental group 30 cases and control group 30 cases,the experimental group to give kidney tonification and detoxification with standard chemotherapy,the control group gives the western medicine chemotherapy alone,both treatment for 4 weeks,more than three consecutive treatment curative effect evaluation.Evaluation index for efficient(CR,VGPR,PR,NR),quality of life score,symptom integral,the level of regulatory T cells.Conclusionkidney tonification and detoxification side compared with the control group had a higher efficient,and with the pure western medicine chemotherapy group no statistical difference,and into the group more than most of the patients received PR curative effect,explain kidney tonification and detoxification parties cooperate with chemotherapy and chemotherapy alone efficient fairly;Kidney tonification and detoxification party group to improve quality of life score,the difference compared with the control group(P<0.05),the kidney tonification and detoxification in order to improve the quality of myeloma chemotherapy patients survival;For the symptoms integral,kidney tonification and detoxification party group than the control group can reduce symptoms integral,and also show statistically significant.Prove that kidney tonification and detoxification may have attenuated synergies to chemotherapy,but is yet to be further study.Not only that,this study also found that the kidney tonification and detoxification group compared with control group,can lower levels of regulatory T cells,but not statistically significant,kidney detoxification tonification and can improve the patients' regulatory T cells,2 experimental research:(1)Using QT-PCR method to detect the mRNA expression of GRP78 amplification genesMethodsthe preparation of drug-containing serum and develop myeloma cell line RPMI8226,the logarithmic phase cells,adjust cell number is 1 x 105/ml,adding adjusted cell suspension 100 ul 96 hole in the enzyme label plate,each group of eight parallel holes,each plate time determination,have set up a list of blank control group,the enzyme label plate placed at 37 ? and 5%C02 concentration,after cultivation in the box under the condition of saturated humidity,respectively,to join different concentrations of medicated serum(blank control group,kidney detoxification group,kidney detoxification + DDT,DDT)different time after the training,respectively,after the drugs involved in multiple myeloma cells of RNA,the reverse transcription reaction cDNA,then carries on the QT-PCR reaction,detecting partner molecular gene mRNA expression of GRP78 augmentation,get the data.Conclusion:by using QT-PCR to detect kidney tonification and detoxification party drug-containing serum couples molecular GRP78 amplification gene mRNA expression,the effect of proof kidney tonification and detoxification pill could increase the expression of GRP78 gene,it follows that the kidney tonification and detoxification party is likely to be one of the myeloma resistance mechanism of via increased GRP78 gene,the endoplasmic reticulum stress,did not fold proteins(UPR)activation,which mediates the myeloma cell apoptosis.(2)using Western blot method to detect Akt,FOX03a phosphorylation and Bim protein expression levels The dose effectMethodsbaht myeloma cells RPMI8226 adding different concentrations of kidney detoxification party drug-containing serum(0,5,10,20 umol/h)treatment for 24 hours,the preparation of protein samples,in 96-well plates respectively protein standard solution,concentration of 0,1,2,4,6,8,9,10 times 20 ml of water,plus 5 times dilution coomassie brilliant blue dye solution 200 ml.In 96-well plates to join protein sample dilution or concentrate in turn 10-20 ml and 5 times dilution of coomassie brilliant blue dye solution 200 ml.On the determination of protein content,carries on the Western blot electrophoresis,transfer film,antibody incubation,immune response,imaging,and then to get the data and image.Conclusionthe results showed that the kidney detoxification can cut AKt,Fox03,the expression of BIM,and assumes the concentration-response relationship,suggesting that the kidney detoxification may by intervening AKt/Fox03/BIM signal transduction,which interfere with the cell survival pathway PI3K/AKt,antagonism NF-KB,help promote myeloma cells apoptosis,resistance to the effect of multiple myeloma.conclusionThis study found that the kidney detoxification in combination with chemotherapy,chemotherapy can improve the patients efficiently,there may be attenuated synergistic effect,can improve the quality of survival,improve patients without incident.Have a kidney detoxification.Experiment shows,can intervene in the regulatory T cells in patients with kidney detoxification,and may strengthen the body's immune monitoring on myeloma cells,immune regulation,its effect on promoting myeloma cell apoptosis;Kidney tonification and detoxification detoxification group after medicated serum role,GRP78 molecular chaperone gene MRNA expression level,it follows that the kidney detoxification party one of myeloma resistance mechanism may be through the increase of GRP78 gene,the endoplasmic reticulum stress,did not fold proteins(UPR)activation,which mediates the myeloma cell apoptosis;Kidney detoxification can cut AKt,Fox03,the expression of BIM,and assumes the concentration-response relationship,suggesting that the kidney detoxification may by intervening AKt/Fox03/BIM signal transduction,which interfere with the cell survival pathway PI3K/AKt,antagonism NF-KB,help promote myeloma cells apoptosis,resistance to the effect of multiple myeloma.This study preliminarily shows the kidney tonification and detoxification detoxification clinical curative effect and mechanism of action,but still need to increase sample size for further study,in addition,the kidney tonification and detoxification detoxification targets and comprehensive mechanisms,remain to be further research.