人参研究GINSENG RESEARCH 2022年第1期
(5-FU)曲笑锋1，石菊1，李铁锡2，于艳辉1，刘松枝2，唐晓桐1，徐建1，鄢必新1*
(1.吉林修正药业新药开发有限公司，长春130103;2.吉林天力泰药业有限公司，长白朝鲜族自治县134400)
摘
要:(5-FU)NF-kb p65/p38,。①BALB/c ,,6,、5-FU (70mg/kg)、1(0.5g/kg)、3(1.5g/kg)、1+5-FU (0.5g/kg+70mg/kg)、3+5-FU (1.5g/kg+70mg/kg),NK ;②Western Blot NF-kb p65p38。①3+5-FU IgM、IgG、TNF-α(P <0.05,P <0.001),1+5-FU IgM、IgG (P <0.01);5-FU ,3+5-FU IgG (P <0.01),1+5-FU IgG (P <0.001)。NK 81.5%、41.1%、159.6%、96.2%、86.3%、120.6%194.2%。②5-FU 1、3+5-FU NF-kb p65/p38。5-FU NK ,NF-kb p65/p38,Bcl-2,,。
关键词:;5-FU;;;NK ;NF-kb p65;p38
Abstract:Objective To investigate the effect of FuFangWanNianQing Capsule combined with fluorouracil
injection (5-FU)on the level of NF-kb p65/p38and immunomodulatory in gastric cancer bearing mice,and to provide experimental basis for clarifying the pharmacodynamic mechanism.Methods ①To establish the subcutaneous transplanted tumor model of gastric cancer in BALB/c mice.In addition to the control group,the tumorigenic mice were randomly divided into 6groups:model group,5-FU group (70mg/kg),1time clinical do of FuFangWanNianQing Capsule group (0.5g/kg),3times clinical do of FuFangWanNianQing Capsule group (1.5g/kg),5-FU +1time clinical do of FuFangWanNianQing Capsule group (0.5g/kg+70mg/kg),5-FU+3times clinical do of FuFangWanNianQing Capsule group (1.5g/kg+70mg/kg),and detect the changes of NK cell activity and rum enzymes in tumor bearing mice after treatment;②The expressions of NF-kb p65and p38proteins in tumor tissues of tumor bearing mice were detected by Western blot.Results ①The activities of IgM,IgG and TNF-αin the rum of tumor bearing mice of the 5-FU +3times clinical do of FuFangWanNianQing Capsule group decread in varying degrees (P <0.05,P <0.001),and the activities of IgM and IgG in the rum of tumor bearing mice of the 5-FU+1time clinical do of FuFangWanNianQing Capsule group decread significantly (P <0.01);Compared with the 5-FU group,the rum IgG activity of tumor bearing mice in the 5-FU+3times clinical do of FuFangWanNianQing Capsule group decread significantly (P <0.01),and the rum IgG activity of tumor bearing mice in the 5-FU+1time clinical do of FuFangWanNianQing Capsule group decreas
ed significantly (P <0.001);The killing rates of NK cells in each experimental group were 81.5%,41.1%,159.6%,96.2%,86.3%,120.6%and 194.2%.②5-FU group,5-FU+3times clinical do of FuFangWanNianQing Capsule group and 5-FU+1time clinical do of FuFangWanNianQing Capsule group could down regulate NF-kB p65/p38level.Conclusion FuFangWanNianQing Capsule combined with 5-FU can improve body immunity by enhancing NK cell killing activity,and its anticancer mechanism may be through reducing NF-kB p65/p38level,inhibit Bcl-2activation,so as to inhibit tumor cell proliferation and promote tumor cell apoptosis.
Keywords:FuFangWanNianQing Capsule;5-FU;combined application;gastric cancer;NK cell activity;NF-kb p65;p38
Immunomodulatory effect and anticancer mechanism of
FuFangWanNianQing Capsule combined with fluorouracil injection(5-FU)
on gastric cancer bearing mice
基金项目:,:20190404002YY。。
作者简介:,,,,。
*通信作者:,,,。E-mail:***************.
QU Xiao-feng 1,SHI Ju 1,LI Tie-xi 2,YU Yan-hui 1,LIU Song-zhi 2,TANG Xiao-tong 1,XU jian 1,YAN Bi-xin 1*
(1.Jilin Xiuzheng Pharmaceutical New Drug Development Co.,Ltd.,Jilin 130103;2.Jilin Tianlitai Pharmaceutical Co.Ltd.,Jilin 134400)
DOI:10.19403/jki.1671-1521.2022.01.006
28
、、、、、、、、、。,。、、
,。
,
NF-kb p65/p38,,
。
1材料
1.1
BALB/c,,18~22g,
,:SCXK() 2020-0002。
1.2
:
,:20190701;(5-FU):
,:190604;10×:So⁃larbio,:D1060-500;5×Tris-, Solarbio,:T1070-500;5×loading buffer: Biosharp,:660272;1×PBST:Solarbio, :T1070-500,P1031-500;ECL
:,:20191121; Page Ruler Prest:thermofisher,:00318120;β-actin:BOSTER,:BA2305;P38MAPK:CST,: 8690;NF-kb/p65,CST,:8242;: BOSTER,:BA1054;:BOSTER, :BA1050;PBS:HyClone,:AE29449011;R
P⁃MI1640:HyClone,:AE29163435;Gey's
:,:L14M11G113128;LDH :,:20201228; IgG:,: 20201228;IgM:, :20201228;TNF-α:
,:20201229。
1.3
SMP500-18272-LSIO;JA1103N
;HHS;;Trans-blot SD cell;Gel imaging system;MCO-18AIC
。
2方法
2.1
BALB/c84,7,12, ,、、5-FU 、1、
3、1
+5-FU(0.5g·kg-1+70mg·kg-1)、
3+5-FU(1.5g·kg-1+ 70mg·kg-1),14d。
2.2
,
,,,
,,,0.9%
, 2.0×106/mL,
。
2.3
,
7d,6×106mL,0.2mL
[1-5]。24h,、
0.9%10mL·kg-1,1, 5-FU1/14d,5-FU
、5-FU,
5-FU,,1。
14d。
2.4Western Blot[6-7]
50mg,800μL ,,4℃
1h,12000rpm,20min,,
,-80℃。,50μg ,5×,98℃
5min,10s。10%SDS-
,80V,20min,, 100~120V,80min。,20V,50min。
5%PBST1h, ,(3%BSA PBST1∶1000
),4℃,。,PBST,10min/。
(5%1∶1000),, 45min。,PBST,10min/。A
B1∶1,,
NF-kb p65、p38。2.5[8]
,-80℃,
。G(IgG)、M(IgM)、α(TNF-α),
。
2.6NK[9-11]
2.6.1
,, PBS,。
70μm。
1500,5min Gey's, 3min。
,PBS。
2.6.2NK
K562。100μL K562(2.5×106/mL) 96。100μL
(1×107/mL)。
。:
:K562100μL+100μL
:K562100μL+RPMI1640100μL
:K562100μL+LDH() 100μL
37℃、5%CO24h LDH
LDH。NK
:
人参研究GINSENG RESEARCH 2022年第1期
NK 81.5%、41.1%、159.6%、96.2%、86.3%、120.6%194.2%,2。
表2
对小鼠NK 细胞杀伤率的影响(x±S,n=12)
3.2
NF-kb p65/p38
,,5-FU 1、3+5-FU NF-kb p65/p38;1、3NF-kb p65/p38,1。
1NF-kb p65、p38NK (%)=OD test -OD min OD max -OD test
×100%
OD test ,OD min ,OD max 。2.7SPSS 19.0,±,P <0.05;Western Blot ,Image J、Graphpad Prism 。3实验结果
3.13.1.1,,IgM、IgG、TNF-α(P <0.05,P <0.001);,3+5-FU IgM、IgG、TNF -α(P <0.05,P <0.001),1+5-FU IgM、IgG (P <0.01);5-FU ,3+5-FU IgG (P <0.01),1+5-FU IgG (P <0.001),1。
3.1.2NK 表1对胃癌MFC 荷瘤小鼠血清中IgM、IgG 和TNF-α活力的影响(x±S,n=12)
IgM(μg/mL)IgG(mg/mL)TNF-α(ng/L)—4163.95±405.62
10.6±5.083.3±12.2—
7999.39±373.41▲▲▲
26.2±9.4▲▲▲
96.6±11.7▲5-FU 70mg/kg 4291.11±584.53***27.8±3.583.0±23.510.5g/kg 3094.20±300.69***14.5±7.3*84.3±33.13 1.5g/kg 3861.00±449.76***15.9±4.4*80.4±7.7*1+5-FU 0.5g/kg+70mg/kg 4053.04±374.88***13.2±6.7**△△△83.3±14.63+5-FU 1.5g/kg+70mg/kg
3986.08±388.85***
11.2±4.1**△△
76.8±15.5*
注：与空白对照组比较,▲P <0.05,▲▲▲P <0.001；与模型对照组比较,*P <0.05,**P <0.01,***P <0.001；与5-FU 对照组比较,△△P <0.01,△△△P <0.001。
NK (%)—81.5±2.0—41.1±6.25-FU 70mg/kg 159.6±4.510.5g/kg 96.2±1.9
3 1.5g/kg 86.3±2.01+5-FU 0.5g/kg+70mg/kg
120.6±7.33+5-FU 1.5g/kg+70mg/kg
194.2±2.5
30
人参研究GINSENG RESEARCH2022年第1期
4讨论为国奉献的诗句
,, ,,,
,50%1
,270%[12]。
,
,。
,
,、
[13]。
、、、,
、、,、、
,、,
,、,。NK
(antibody-dependent cell-me⁃diated cytotoxicity,ADCC)[14],
,NK
。BALB/c
,
[15,16],,
NK, IgG、IgM,
TNF-α,
,
。
金鱼怎么画>抢不到票怎么办
香港关税6:
、、、
、、[17,18]。
NF-kb;MAPK
,JNK、ERK、p38。, p38、、,
。p38
,p38
Bcl-2Bax,
[19-22]。,
NF-kb p65/p38,
NF-kb p65/p38MAPK,p38
她力量Bcl-2,
安全语录经典
。,5-FU
NF-kb p65/p38,Bcl-2,
,。
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