| Background and objective:Lymphoma is a malignant tumor of lymph nodes and/or extranodal lymph nodes.Pathological findings showed a large proliferation of neoplastic lymphocytes with different degrees of differentiation and maturity had invaded each part or tissue of the whole body.The main clinical symptoms are painless,progressive lymphadenopathy,and often accompanied by fever,night sweats,emaciation,liver and spleen large,late anemia,cachexia and other clinical manifestations.In recent years,the study found that the incidence of lymphoma is increasing year by year,for all cancer patients in the proportion is about 5%,of which non Hodgkin’s lymphoma(Non-Hodgkin Lymphoma,NHL)accounted for about 89.1%of malignant lymphoma,About 7 times as much as Hodgkin lymphoma(HD).The high incidence areas of non Hodgkin lymphoma are distributed in Western Europe,the United States and the Middle East,while China,Japan and so on are low incidence areas.From the point of view of the trend,Hodgkin lymphoma incidence tends to be stable,while non Hodgkin lymphoma incidence is on the rise in developed countries.The disease can occur at any age,but the peak age of onset is 31-40 years,including NHL peak slightly forward.The sex ratio of men and women is:2-3:1.It is generally believed that it may be related to genetic mutations,as well as viruses and other pathogens,infections,radiation,chemicals,and autoimmune diseases.Malignant lymphoma is a kind of tumor with considerable heterogeneity,although occurs in the lymph node,but due to the distribution characteristics of the lymphatic system,the lymphoma is a systemic disease,can affect almost any tissue and organ in the whole body.Therefore,the clinical manifestations of malignant lymphoma both have some common characteristics,and vary greatly according to different pathological types,site and range of invasion.In China,286 out of every 100 thousand people have cancer.The incidence of malignant lymphoma is increasing and more than 25 thousand new cases are reported each year so it has become a major tumor threatening the health of the people.While in Europe,the Americas and Oceania,the incidence of malignant lymphoma is 18/10 million,slightly higher than the sum of all kinds of leukemia.In the United States,more than 30 thousand new cases of malignant lymphoma are added each year.The mortality rate of malignant lymphoma is 1.5/10.0 million in China,and is the 10-14 place in the ranking of the number of deaths from all malignancies,which is similar to leukemia.T cell lymphoma is often characterized by extranodal lesions,and necrosis and apoptosis are common in biopsy tissues.Compared with B cell lymphoma,T cell lymphoma has poor efficacy and poor prognosis,so to further study the molecular mechanism of T cell lymphoma and the prognosis more accurate assessment is particularly important.Anaplastic large cell lymphoma(anaplastic large cell lymphoma,ALCL)is a type of independent classification established by WHO in 2008.It is a relatively rare,aggressive mature T cell lymphoma,divided into ALK positive and ALK negative.the unmber of ALK positive anaplastic large cell lymphoma accounts for 3%adult and 10%-20%children non Hodgkin lymphoma,which age of onset before the 35 years old and males are more frequent.2 types of Anaplastic large cell lymphoma(ALK negative and positive)are similar in morphology,but the negative type lack anaplastic large cell lymphoma kinase ALK.T cell associated markers and cytotoxic granule associated proteins were expressed in most cases.ALK negative anaplastic large cell lymphoma is generally composed of larger and more pleomorphic,more obvious nucleolus cells.Compared with ALK positive anaplastic large cell lymphoma patients,the negative type prognosis is poor,the 5 year survival rate is about 48%,,and 5 year progression free survival is about 36%.According to the national comprehensive cancer network(the National Comprehensive Cancer Network guidelines),the treatment of anaplastic large cell lymphoma with chemotherapy,radiotherapy,surgery,assisted by stem cell transplantation combined treatment.In the early stage of the disease,chemotherapy should be followed by local radiotherapy,while chemotherapy should be used in the middle and late stages.However,because of the side effects of radiotherapy and chemotherapy,many patients can not tolerate it.Therefore,it is one of the main research contents to seek antitumor drugs with high efficiency and low toxicity at present.Chinese medicine treatment of tumors in China has a long history.It has been proved by clinical application that Chinese herbal medicine for treating malignant tumor not only has the effect of reducing toxicity and synergism,sensitivity to chemotherapy and radiotherapy;but also can prevent tumor recurrence and metastasis,blocking precancerous lesions and antitumor effect.It can also improve the immunity of the body and not destroy the normal cells of the body when it is used for the treatment of tumors.This is the effect that many chemotherapy drugs do not have;At the same time,Chinese herbal medicine has many advantages,such as no obvious side effects,wide distribution of drug sources,low price,suitable for long-term use and so on.It is one of the effective methods in the comprehensive treatment of malignant tumors.Prunella vulgaris L has benifits on liver,eye,loosing knot,reducing swelling,and analgesic effect.It is used for the pain of the eyes,the shaming of the eyes,the dizziness of the headache,the scrofula,the galls and the swelling of the breast.At present,this drug is used to treat the diseases such as goiter,lymphatic tuberculosis,mammary gland hyperplasia,hypertension,tuberculosis,acute jaundice and bacterial dysentery.Prunella vulgaris L contains rich chemical constituents,the main components are:triterpenoids,steroid,flavonoids,coumarin,organic acids,sugars,volatile oils,etc.There are many reports about the anti-tumor effect of Prunella vulgaris,such as application selfheal treatment of thyroid cancer,lymphatic sarcoma,carcinoma of the parotid gland,tonsil cancer,nasopharyngeal carcinoma,breast cancer,cervical cancer,and liver cancer and so on for a long time.At present,human tumor research has reached the level of genomics,proteomics,metabolomics,epigenetics.Genome sequencing of the whole genome marks the arrival of the post genome era;people are more and more concentrated interest in protein research,thus proteomics research also has a rapid development.Research on proteomics in cancer prevention plays an important role in the pathogenesis of tumor,tumor early diagnosis of cancer,clinical treatment and prognosis of tumor and anti tumor drug development and showed bright prospect.Metabonomics(metabonomics/metabolomics)is thought to follow the research of genomics and proteomics and is part of the biological system.The research methods is quantitative analysis of all metabolites in organisms,and find the relative metabolites with physiological and pathological changes,Most of the research objects are small molecules with a relative molecular mass of less than 1000.Epigenetics is a branch of genetics that studies genetic changes in gene expression without changing the nucleotide sequences of genes.Epigenetic phenomena are numerous,such as DNA methylation(DNA,methylation)(genomic imprinting)genomic imprinting,maternal effect(maternal effects),gene silencing(gene silencing),nucleolar dominance,transposable elements activation and RNA editor(RNA editing).As the material basis of life,protein is the main executor of cell metabolism and regulation.It is also a target of many pathogenic factors and most drugs.Because of any disease before and after before the occurrence of pathological changes and clinical treatment,The components and the number of intracellular protein will change,so,in theory,through the dynamic detection of proteins that can detect the changes of protein quality and quantity before and after the disease early and clinical treatment as early disease the diagnosis,treatment and prognosis prediction index.We have been studied the effect of extract from Prunella vulgaris L on different lymphoma cell lines for 11 years,in order to confirm the extract may have a certain effect on T cell lymphoma,this study selected Karpas299、Jurkat、HUT78 cells as the research object.First,We observed the antiproliferative effect of extract from Prunella vulgaris L on these T cell lymphoma,and found the karpas299 cell was the best.Then we study the anti-tumor effect in vitro by TMT identification of the extract from Prunella vulgaris L,differences in relative quantitative proteomics and metabolomics technology screening and identification of the extracts from the extract from Prunella vulgaris L Karpas 299 lymphoma cell line protein molecules and difference metabolites,find the possible mechanism of the extract from Prunella vulgaris L Karpas lymphoma cell lines.TNFα、The transcription factor c-Jun、cyclinD3 are proteins expression proteomics found the related gene which can induce or inhibit a group of potential tumors,in order to further verify the efficacy of the extract from Prunella vulgaris L,so we use BALB/c to construct Karpas in nude mice lymphoma animal model,further research on the effect of the extract from Prunella vulgaris L,and expression in in vitro and in vivo experiments were used to study proteins by the Western Blot.Part 1 Effects of the extract from prunella vulgaris L on the biological characteristics of Karpas 299 cells Methods:(1)Cultured Karpas299、Jurkat、HUT78 lymphoma cell lines.(2)The CCK-8 method was used to determine IC50,respectively,the extract from prunella vulgaris L and the vincristine were given,and the ratio concentration gradient was set,and the growth inhibition rate of the Karpas 299、Jurkat、HUT78 cells were studied.(3)Determination of cell vitality and the observation of cell morphology by inverted microscope.(4)The extract from prunella vulgaris L have no distinct cell cycle specificity to karpas299 cells.(5)The cell apoptosis was detected by flow cytometry,and analyzed the difference between the extract from prunella vulgaris L,the vincristine group,and the combined group.Results:(1)The different concentrations of the extract from prunella vulgaris L on the karpas 299、Jurkat、HUT78 cells after 48h,the karpas significant inhibition of proliferation,the extract from prunella vulgaris L respectively with 100 ug/ml,50 ug/ml,25 ug/ml,12.5 ug/ml,6.25 ug/ml concentration processing Krapas 299、Jurkat、HUT78 cells for 48 h,karpas 299 cells growth inhibition rate were 79.41±2.56%、61.47.±1.90%、44.52±1.73%、30.38±1.04%、8.23±0.98%.Its proliferative IC50 is 33.2±1.8ug/ml.With the increase of the concentration of the extract,the inhibition rate is higher and higher,and there is a dose and time dependence within a certain concentration range.Vincristine also has obvious inhibitory effect on the karpas 299 cells,different concentrations of vincristine on the karpas 299 cells after 48h,drug concentration is 0.008 ug/ml,0.004 ug/ml,0.002 ug/ml,0.001 ug/ml,0.0005 ug/ml,respectively for karpas cell growth inhibition rate was 79.41±2.56%、61.47.±1.90%、44.52±1.73%、30.38±1.04%、8.23±0.98%,its proliferation IC50 for 0.003±0.0004ug/ml.Jurkat cells growth inhibition rate were 81.2±2.73%、62.35±1.89%、44.38±1.61%、29.56±1.04%、7.45±0.96%.Its proliferative IC50 is 46.3±2.1 ug/ml.With the increase of the concentration of the extract,the inhibition rate is higher and higher,and there is a dose-dependence within a certain concentration range.Vincristine also has obvious inhibitory effect on the Jurkat cells,different concentrations of vincristine on the Jurkat cells after 48h,drug concentration is 0.008 ug/ml,0.004 ug/ml,0.002 ug/ml,0.001 ug/ml,0.0005 ug/ml,respectively for Jurkat cell growth inhibition rate were 89.26±2.65、50.19±2.42、41.05±1.87、30.68±1.35、19.73±1.11,its proliferation IC50 for 0.006±0.0009ug/ml.HUT78 cells growth inhibition rate were 78.20±2.49%、60.35±1.95%、45.38±1.81%、32.56±1.04%、9.01 ±0.97%.Its proliferative IC50 is 47.1 ±2.2ug/ml.With the increase of the concentration of the extract,the inhibition rate is higher and higher,and there is a dose-dependence within a certain concentration range.Vincristine also has obvious inhibitory effect on the HUT78 cells,different concentrations of vincristine on the HUT78 cells after 48h,drug concentration is 0.008 ug/ml,0.004 ug/ml,0.002 ug/ml,0.001 ug/ml,0.0005 ug/ml,respectively for HUT78 cells growth inhibition rate were 88.47±2.73、50.37±2.35、33.84±1.62、25.66±1.48、18.79±1.59,its proliferation IC50 for 0.007±0.0008ug/ml.Both drugs had significant dose dependence on cell inhibition.(2)The IC50 of extract from prunella vulgaris L and vincristine injection treated karpas 299 cells was 33.2ug/ml,0.003ug/ml.The difference between the two drugs in the experimental group and the blank control group was statistically significant,P<0.001.(3)The extract from prunella vulgaris L and vincristine treated karpas 299 cells 48h,and the concentration of IC50 was 33.2ug/ml,0.003ug/ml.The blank group G0/G1 phase cells accounted for 67.1±4.2%,G2/M phase cells accounted for 6.2±0.9%,the extract from prunella vulgaris L group G0/G1 phase cells accounted for 61.3±3.9%,G2/M phase cells accounted for 7.6±1.1%,the vincristine group G0/G1 phase cells accounted for 43.6±2%,G2/M phase cells accounted for 26.1±2.5%,and the group G0/G1 cells was 51.8±1.4%.G2/M phase cells accounted for 10.3±1.4%.Compared with the blank group,the G2/M phase cells in vincristine group increased significantly,p<0.01(P=0.0016),the G2/M phase cells in the combined group increased,p<0.05(P=0.038)and the extract from prunella vulgaris L group had no significant difference,p>0.05.(4)The extract from prunella vulgaris L and vincristine treated karpas 299 cells 48h,and the concentration of IC50 was 33.2ug/ml,0.003ug/ml.The apoptosis rate of Karpas 299 cells in blank control group was(1.32±0.14)%,the apoptosis rate of Karpas 299 cells in the extract from prunella vulgaris L group was(38.9±2.95)%,the apoptosis rate of Karpas 299 cells in the vincristine group was(42.8±3.71)%,the apoptosis rate of Karpas 299 cells in the combined group was(66.3±7.52)%,compared with the blank control group.P<0.01(P=0.0093,P=0.0064,P=0.0058).Summary(1)The extract from prunella vulgaris L has a significant anti-proliferation effect on Karpas 299 cells、Jurkat cells、HUT78 cells,and its anti-proliferation effects are time and dose dependent.(2)The antiproliferative effect from Prunella vulgaris extract on Karpas299 cells with no distinct periodic specificity.(3)Early apoptosis of Karpas 299 cells can be induced by the extract from prunella vulgaris L.Part 2 Proteomic analysis of Karpas 299 cells after the trement of prunella vulgaris L and function verification of key protein Methods:(1)Divided into three groups,respectively for the blank group,selfheal extract mu(33.2ug/ml),vincristine group(0.003ug/ml),each group of three repetition,collect extract from prunella vulgaris L respectively(33.2ug/ml),vincristine(0.003ug/ml),Acting on Karpas 299 cells after 48 h,The cells in the cell culture flasks were washed 3 times with a cell collection solution,the number of cells collected per tube was 5.0×107,liquid nitrogen cryopreservation.Protein extraction was performed using 1mM PMSF,2mM EDTA and DTT.(2)The BCA method was used for protein quantification.(3)The bioinformatics analysis of differential protein molecules mainly includes the GO function annotation and the KEGG signal pathway enrichment analysis.(4)The key differential proteins related to the tumor were found,some of which were verified by Western blot.Results:(1)This project uses TMT quantitative proteomics technology to carry out project research,and identified 6209 proteins in human cells.According to the expression multiple,the variation of the expression multiple was more than 1.2 times(the increase was greater than 1.2 times or less than 0.83 times)and P value<0.05 was the standard for the expression of protein.In this study,compared with the control group,there were 55 up-regulated proteins,such as Protein lifeguard 3、Annexin A1、CD160 antigen、Perforin 1、Myomegalin、Prothrombin、Fibronectin type III domain-containing protein 3B、Extracellular matrix protein 1、Filamin-B、Interferon-induced 35 kDa protein、Smoothelin、Thymidine phosphorylase、Vesicular integral-membrane protein VIP36、Cathepsin D、TNFα、Nicotinamide phosphoribosyl transferase and so on,and 77 down regulated proteins,such as Collagen type I alpha 1、Synaptophysin、V excision repair protein RAD23 homolog B、Myelin proteolipid protein、Synapsin-1、Antileukoproteinase、E3 ubiquitin-protein ligase RNF123、Stathmin、GO2-q chimeric G-protein、Transcription factor c-Jun、Creatine kinase B-type、Putative heat shock protein HSP 90-beta 4、Calcium/calmodulin-dependent protein kinase type Ⅱ subunit alpha、Protein FAM210A、MSTP067、cDNA FLJ78114、High mobility group protein B2、Cyclin-dependent kinase inhibitor 2A、CyclinD3 and so on.After recombinant expression plasmid transformation,protein induced expression,purification and recombinant protein enrichment,SDS-PAGE,then we selected TNFa,c-Jun and cyclinD3 as significant difference proteins to carry out functional verification.(2)After 48 administration,The results of SDS-PAGE electrophoresis and Western blot showed that the expression protein was compatible with c-Jun protein by recombinant expression plasmid transformation and protein induced expression,purification and enrichment of recombinant proteins.Western blot results showed that after 48h administration,Prunella vulgaris extract group,vincristine group and combined group reduced p-JAK3,p-STAT3,p-JNK,p-c-Jun,p-AKT,cyclinD3,Bcl-2 and mTOR to different degrees,and increased the expression of TNFa,Bcl-2 and mTOR.Compared with the control group,group of Prunella vulgaris extract with high expression of TNFa,P<0.05(P=0.0217),low expression of p-cJun,P<0.05(P=0.0164),low expression of cyclinD3,P<0.05(P=0.0329);high expression of TNFa protein in vincristine group,P<0.05(P=0.0283),low expression of p-cJun,P<0.05(P=0.0175),low expression of cyclinD3,P<0.05(P=0.0272);high expression of TNFa protein in combination group,P<0.01(P=0.0079),low expression of p-cJun,P<0.01(P=0.0052),low expression of cyclinD3,P<0.01(P=0.0061).Summary:(1)When the extract from prunella vulgaris L treated karpas299 cells 48h,many proteins successfully identified by mass spectrometry with statistical significance,and these proteins involved in apoptosis,suggesting that the anticancer mechanism may be related to this.(2)When the concentration of the extract from prunella vulgaris L was 33.2 ug/mL,the growth inhibition rate for karpas cells was about 50%,suggesting that the extracts of summer grass can significantly inhibit the growth of karpas cells.This experiment found that,up regulation of 219 proteins and down regulation of 321 proteins were successfully identified by mass spectrometry:Up regulation Protein respectively lifeguard 3,(3)After Western blot validation,the extract from prunella vulgaris L may play an anticancer role by downregulating p-JAK3,p-STAT3,p-JNK,p-c-Jun,p-AKT,mTOR,cyclinD3 and Bcl-2 expression,thereby increasing TNFa,Bax,Caspase3 expression.These proteins involved in cell metabolism and suggested that the mechanism of anticancer cells from extract of Prunella vulgaris may be related to JAK3/STAT3、JNK/cJun、AKT/mTOR signaling pathways.Part 3 Metabolomics analysis of Karpas 299 cells after the trement of extract from prunella vulgaris L Methods:(1)Divided into 3 groups,respectively for the blank group,the extract from prunella vulgaris L group(33.2ug/ml),vincristine group(0.003g/ml),each group of 6 repeat,respectively collect Karpas 299 cells in blank group and in selfheal extract(33.2ug/ml),vincristine(0.003 ug/ml)action after 48 h group,The cells in the cell culture flasks were washed 3 times with a cell collection solution,the number of cells collected per tube was 5.0×107,liquid nitrogen cryopreservation.(2)Ultrasonic cracking was used to extract protein.(3)Using Agilent 1290 Infinity LC super high performance liquid chromatography(UHPLC)HILIC chromatographic column separation.Electrospray ionization(ESI)positive ion and negative ion mode are used to detect.After the separation of the sample UHPLC,the mass spectrometry of the Triple TOF 5600 mass spectrometer(AB SCIEX)was performed.(4)Adopt XCMS program for peak alignment,retention time correction and extraction peak area.(5)According to the target C number respectively of metabolites in KEGG database to extract all the target metabolites involved metabolic pathways,and respectively indicate target metabolites in metabolic pathways through the KEGG Mapper software.(6)Metabolite cluster analysis.In the enrichment analysis of the annotation of the KEGG pathway of the target metabolite,the KEGG pathway was used as the unit to the background of all metabolites in each pathway,and the exact test was done by Fisher.Results:(1)The QC samples UHPLC-Q-TOF MS total ion flow graph was overlapped to compare,the result shows that the response intensity and retention time of chromatographic peak overlap basicly,it shows that the variation of instrumental error is smaller during the whole experiment.(2)After Pareto-scaling,the PCA analysis was carried out,and the QC samples in the positive and negative ion mode were closely clustered together,indicating that the experiment of this project was good repeatability.(3)The mechanism of the extract from prunella vulgaris L inhibits Karpas 299 cell proliferation was mainly by participating in multiple metabolic pathways,especially different metabolites in the positive ion mode such as the L-Carnitine、Betaine、L-Palmitoylcarnitine、Taurine、Glycerophosphocholine,and different metabolites in the negative ion mode such as the Stearic acid、Glycerol 3-phosphate、Arachidonic Acid(peroxide free)、Linoleic acid、L-Palmitoylcarnitine、Stearoylcamitine、Taurine、alpha-D-Galactose 1-phosphate、2-Dehydro-3-deoxy-D-gluconate。The mechanism of vincristine inhibits Karpas 299 cell proliferation was mainly by participating in multiple different metabolites in the positive ion mode such as alpha-D-Glucose 1-phosphate、Uridine 5’-monophosphate(UMP)、Uridine 5’-diphosphate(UDP)、Pantothenate,and different metabolites in the negative ion mode such as Linoleic acid、L-Citrulline、Uridine 5’-monophosphate(UMP)、alpha-D-Galactose 1-phosphate、cDNA FLJ56762、Alpha-2-HS-glycoprotein、cDNA FLJ52778、Insulin-like growth factorbinding protein 7、Keratin,type Ⅱ cytoskeletal 2 epidermal and so on.Summary(1)The mechanism of the extract from prunella vulgaris L inhibits Karpas 299 cell proliferation was mainly by participating in multiple metabolic pathways,especially the UDP-D-Galactose、alpha-D-Glucose 1-phosphate and other metabolites,to participate in the swelling of inhibiting tumor cell proliferation.(2)To study the function of these metabolites,the effect of promoting or suppressing the proliferation and apoptosis of tumor cells provides the basis for subsequent experiments.Part 4 The experiment of the extract from prunella vulgaris L in vivo and function verification of key protein Methods:(1)To construct the animal model of Anaplastic large cell lymphoma,we divided the extract from prunella vulgaris L into 3 groups(the high dose group,the middle dose group and the low dose group)at first and found the middle dose group was the best,then we studied 6 groups which is divided into the blank control group,the low dose group,the middle dose group and the high dose group,vincristine group,and the combined group(vincristine combined with the middle dose of extract from prunella vulgaris L).(2)Daily observation of the growth of the tumor mice in each group,weighing on the next day,and recording the weight changes of the mice in each group.(3)The size of the tumor was measured.(4)Histopathology HE staining and immunohistochemistry.(5)Western blot detection of p-JAK3、p-STAT3、p-JNK、p-c-Jun、p-AKT、mTOR、cyclinD3、TNFα、Bcl-2、Bax、caspase3 protein expression in tumor tissues metabolites in each pathway,and the exact test was done by Fisher.Results:(1)The rate of tumor formation in Karpas 299 lymphoma animal model was 100%.(2)The extract from prunella vulgaris L can inhibit Karpas 299 lymphoma.Compared with the blank control group,the volume of tumor tissue decreased significantly in three dose groups in the pre experiment,P<0.01(P=0.0075,P=0.0062,P=0.0071).Compared with the blank control group,the average tumor weight of the low and high dose group decreased,P<0.05(P=0.0283,P=0.0196),and the average tumor weight of the middle dose group decreased significantly,P<0.01(P=0.0044),the tumor suppressor rates of each group were 27.3%,40.1.5%and 31.5%respectively.Compared with the blank control group,the volume of tumor tissue decreased significantly in three dose groups in the formal experiment,P<0.01(P=0.0094,P=0.0091,P=0.0077),and the average tumor weight of the three dose groups decreased significantly,P<0.01(P=0.0072,P=0.0055,P=0.0068),the tumor suppressor rates of each group were3 8.9%、39.5%、39.1%.(3)It suggested that T cells labeled CD3,CD5,and CD7 were all expressed in varying degrees by immunohistochemical staining.CD30 was all diffuse cell membrane positive.ALK was positive in the blank control group and the extract from prunella vulgaris L group,and the other groups were negative.The proliferation index of Ki67 was 60-80%in the blank control group and the vincristine group,but the proliferation index of Ki67 was 30-40%in the other groups.Compared with the blank control group,the expression of c-Jun in each group decreased,P<0.05(P=0.0291).There was no significant difference between the drug groups,P>0.05.The expression of cyclinD3 in each group was reduced,P<0.05(P=0.0406).The cyclinD3 in the combined group was significantly lower,P<0.01(P=0.0054).The expression of TNFa was significantly higher in each drug delivery group,but the expression of TNFa increased in the low dose group,the extract from prunella vulgaris L group,P<0.05(P=0.0316),vincristine group and combined group increased significantly,P<0.01(P=0.0082).(4)Through Western blot expression,Compared with the blank control group,the expression of c-Jun in each group decreased,P<0.05(P=0.0291).The expression of TNFa of the extract from prunella vulgaris L group increased,P<0.01(P=0.0047),p-cJun decreased,P<0.05(P=0.0265),cyclinD3 decreased,P<0.05(P=0.0329);The expression of TNFa of the vincristine group increased,P<0.01(P=0.0051);p-cJun decreased,P<0.01(P=0.0078),cyclinD3 decreased,P<0.01(P=0.0096);The expression of TNFa of the combined group increased,P<0.01(P=0.0015),p-cJun decreased,P<0.01(P=0.0012),cyclinD3 decreased,P<0.01(P=0.0053).Summary(1)The extract from prunella vulgaris L can inhibit Karpas 299 lymphoma and the middle dose group was the best.(2)The extract from prunella vulgaris L reduced p-JAK3,p-STAT3,p-JNK,p-c,Jun,p-AKT,mTOR,cyclinD3,Bcl-2,and up regulated TNFα,Bax,Caspase3 proteins,and may play an anti-tumor role by acting on the JAK3/STAT3、JNK/cJun and AKT/mTOR signaling pathways.Conclusion 1.The extract from prunella vulgaris L has a significant anti-proliferation effect on Karpas 299 cells,and its anti-proliferation effects are obvious in time and dose dependence.The extract from prunella vulgaris L has no distinct cell cycle specificity to karpas299 cells.Early apoptosis of Karpas 299 cells can be induced by the extract from prunella vulgaris L.2.Up regulation of 219 proteins and down regulation of 321 proteins were successfully identified by mass spectrometry,these proteins involved in cell metabolism,the extract from prunella vulgaris L may play an anti-tumor effect through JAK3/STAT3、JNK/cJun and AKT/mTOR signaling pathways.3.The extract from prunella vulgaris L inhibits Karpas cell proliferation,mainly by participating in multiple metabolic pathways,there were 9 different metabolites in the positive ion mode,and 14 different metabolites in the negative ion mode,to participate in the swelling of inhibiting tumor cell proliferation.4.The extract from prunella vulgaris L can inhibit Karpas lmphoma and may It may also play an anti-tumor effect through JAK3/STAT3、JNK/cJun and AKT/mTOR signaling pathways. |
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