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Thymopolypeptides combined with matrine type alkaloids suppress HBV replication

苦参碱类生物碱联合胸腺肽抗HBV作用研究



全 文 :2016
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Vol41,No.7 April,2016

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ThymopolypeptidescombinedwithmatrinetypealkaloidssuppressHBVreplication
LIUXiaoqiong1,2,SHENHonghui2,CHENJiaxin2,BAIZhaofang2,WANGJiabo2,XIAOXiaohe2
(1.PharmacyColege,JiangxiUniversityofTraditionalChineseMedicine,Nanchang330004,China;
2.ChinaMilitaryInstituteofChineseMedicine,302MilitaryHospital,Beijing100039,China)
[Abstract] Toinvestigatetheantiviralefectofthymopolypeptidescombinedwith4kindsofmatrinetypealkaloidsonHepG2215
cels,oxymatrine,sophocarpidine,sophocarpine,andsophoridine(atconcentrationof02mmol·L-1respectively)wererespectively
combinedwiththymopolypeptides(0025,01g·L-1),andafter48hand72htreatmentonHepG2215cels,thecelsandsu
pernatantswerecolectedThecelsactivityinvariousgroupswasdeterminedbyCCK8methodtoevaluatethetoxicefectsofthe
drugsonHepG2215celsEnzymelinkedimmunosorbentassay(ELISA)wasusedtodetermineHBeAgandHBsAglevelsincelu
larsupernatantsHBVDNAlevelsincelularsupernatantsandcelswerequantifiedwithfluorogenicquantitativePCRmethod;andthe
expressionlevelofIFNαinsupernatantswasdetectedwithCBAmethodTheresultsindicatedthatsinglethymopolypeptidesat0025
04g·L-1hadnotoxicitytocelsThymopolypeptidesinthisconcentrationrangecombinedwith02mmol·L-1matrinetypealka
loidsalsohadnotoxicitytocelsAntiHBVactivityofdrugcombinationwasbeterthanthatofalkaliorthymopolypeptidesaloneThy
mopolypeptidesat0025g·L-1hadbeterinhibitoryefectthanthymopolypeptidesat01g·L-1onintracelularHBVDNAexpres
sion,buttheinhibitoryefectonsupernatantHBeAglevelwasonthecontraryAntiHBVactivitywassimilarbetweenalkaloidscom
binedwith01g·L-1andalkaloidscombinedwith0025g·L-1TherewasnostatisticaldiferenceinantiHBVefectbetweenvari
·5721·
2016
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Vol41,No.7 April,2016
ouscombinedgroups(P<005)Ingeneral,72hantiHBVefectwasbeterthan48hantiHBVefect(P<005)Theexpression
ofIFNαwasincreasedafterdrugcombination,withpositivecorelationtothechangesofotherfourindicators(P<005)Inconclu
sion,oxymatrine,sophocarpidine,sophocarpineandsophoridinecombinedwiththymopolypeptidescouldinhibitHBsAgandHBeAgse
cretioninHepG2215celsandHBVDNAreplication,andfurtherpromotetheantiviralefectbypromotingtheexpressionofIFNα
[Keywords] thymopolypeptides;matrinetypealkaloids;HepG2215cels;hepatitisBvirus
doi:10.4268/cjcmm20160719
  
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·7721·
2016
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Vol41,No.7 April,2016
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baWXYžŸ®|$ˆ¿…†÷ãä.
¤

>kÚ

„•$,ñ›Ò­Ø



})Ma
\Âj]Û
[12],
§j]ÛB6zôOh0ž
[13]。
çƒ$m±TݾĘ¾Ma\]
IFNα±
·8721·
Mfuè

TݾĘ¾UVWXYô
HBV
0ž«¬
    
A,B
—`•
48,72h
ֻ
HBVDNA
9ˇ
;C,D
—`•
48,72h
a8
HBVDNA
9ˇ
;E,F
—`•
48,72hHBsAg
9ˇ
;G,H
—`•
48,72
hHBeAg
9ˇ

UVž,


.02mmol·L-1
TݾĘ¾UV
0025g·L-1
WXY

UVž,


.02mmol·L-1
TݾĘ¾
UV
01g·L-1。
ÑU´WXYñò
1)P<005,
ÑU´Â˜¾ñò
2)P<005。
q
3 
U´~,¬UVž,±ô
HBV
Éæñò
(n=5)
Fig3 AntiHBVactivityofsingledruggroupsandcombinationgroups(n=5)
·9721·
2016
y

z ³
41
ų

}
Vol41,No.7 April,2016
*
1 
,˜V±*¦µ
Table1 Thesynergisticefectofdrugcombination
˜¾
WXY
/g·L-1
ֻ
HBVDNA
a8
HBVDNA HsAg HeAg
48h 72h 48h 72h 48h 72h 48h 72h
t_Tݾ
0025  + + -  + + +
01  + + +    
Tݾ
   0025  + + + + - - -
01  +  + + +  
Êæ¾
   0025  + + + - + - -
01 + +  + + +  
Êâ¾
   0025  + + + - +  
01 + +  +  +  
  
å
:-
aô0ž
;+
€Å0ž
;*¦0ž。
*
2 
TݾĘ¾UVWXYì
HepG2215
Zaֻ
IFNα*s¢%±òó(珋x±s,n=5)
Table2 TheefectofthymopolypeptidescombinedwithmatrinetypealkaloidsonexpressionofIFNαinHepG2215cel(珋x±s,
n=5) ng·L-1
`
48h 72h
ìH
246±018 287±018
0025g·L-1
WXY
332±075 411±090
01g·L-1
WXY
311±038 501±131
02mmol·L-1
t_Tݾ
364±102 363±123
02mmol·L-1
t_Tݾ
+0025g·L-1
WXY
577±1201,3) 940±1921,2)
02mmol·L-1
t_Tݾ
+01g·L-1
WXY
373±125 748±1721,2)
02mmol·L-1
Tݾ
376±114 384±106
02mmol·L-1
Tݾ
+0025g·L-1
WXY
608±1171,3) 854±1201,2)
02mmol·L-1
Tݾ
+01g·L-1
WXY
419±084 700±2181,2)
02mmol·L-1
Êæ¾
362±080 348±058
02mmol·L-1
Êæ¾
+0025g·L-1
WXY
620±0751,3) 804±1311,2)
02mmol·L-1
Êæ¾
+01g·L-1
WXY
407±107 760±1541,2)
02mmol·L-1
Êâ¾
321±030 506±082
02mmol·L-1
Êâ¾
+0025g·L-1
WXY
626±0711,3) 806±1001,2)
02mmol·L-1
Êâ¾
+01g·L-1
WXY
38±104 768±1651,2)
  
å

Ñ삱U´WXYñò
1)P<005;
Ñ삱U´Â˜¾ñò
2)P<005;
¥¦Å¾UVž,ýþ±ñò
3)P<005。
*
3 IFNαÑÓâ‰Â±€Å(n=5)
Table3  CorelationbetweenIFNαwithotherindicators
(n=5)
‰Â
r P
ֻ
HBVDNA 0873 0001
a8
HBVDNA 0671 0001
HBsAg 0713 0001
HBeAg 0521 0005
*s

UVWXYýY
,IFNα±*sßøÅ,IFNα
± _¬Óâ‰ÂÛ?€µ

#iWXY¬TÝ
¾Ä˜¾U´püUVž,M{É@¸Ö
IFNα±*sÏøeôOhÉæ±。
WXYÀíwžŸñµmQ‚0±…†$ˆ

}ÉIžÃߥÆ[

@¸çƒ*ú

¥¦Å¾±
WXYì
HepG2215
Za±ô
HBV
É楦

¿9Ë
HBeAg
œ‘


0025g· L-1
WXY±É
潟U´
01g· L-1
WXY

}ÉìŸa8
HBV
DNA
9ˇ


01g· L-1
WXYÉ潟U´
0025g· L-1
WXY

WXY¥¦Å¾¿Zaçƒ
÷±¥¦ÉæMWXY¥¦Ãßìf°Ö‚¥{
±˜Ï祦

’#i

¿òY±…†VS‚ž$

5쥦ÜO¥¦*+±ž,Ãßʋš

:/Ø
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…†«¬*úpžWXYUVj]Ûpê
·0821·
Mfuè

TݾĘ¾UVWXYô
HBV
0ž«¬

¼

Ę$ˆñµm‚-BîdˆÉ
[1416],
°ð
烊çt_Tݾ¬­`âUVž,{ðh·

HBV
±9Ë¢%
[17],
t_TݾM{+9Ë
s,
HBV
±æË

¦§¬êÄ,˜_–ß°Å
Ï*¦ôOh

}ìŸ@“ìêĘêtãÂ
s,µaÊìj]Û±$ˆrɱñµmQ‚0û
üÏé

|ˆ~B[â±oCµ

TݾÄ,¤
UVWXYòê͞Ÿçp…†$ˆ$

}ÉÓì
ñµmQ‚0¬./±kY$ˆÁÉB[âˆ
É
[1820]。
9çƒ$
,2
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Ę¾ýY±ô
HBV
Éæ19€”

冽ŸU
´WXY±Éæ

’#i¿…†ž,$

쟥*
zãj]Ûpê

¼

Ę$ˆ±O$

MaÓ
žTݾÄ,¤UVWXYÖnôOh$ˆ

‰÷bg

TݾĘ¾UVWXYô
HBV
É潟U´ž,

ÓôOhfËM{Ñ\]
IFN
α±*sB。€ü±UVIž-Bòݱ…†«
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ÓôOhfËÁB6Ö[Í
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„q=

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$%«JAÚ

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,2010(9):10.
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merasechainreactionamplificationsystem[J].BiotechnolLet,
2015,37(10):2063.
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[10] SpangeloBL,FarimondDD,ThapaM,etal.Thymosinfrac
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†|

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è

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åƞWX´YUVÔñ`â$ˆñµm‚
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ˆÉ—à
[J].
$%çžA,
,2015(18):58.
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ëðÕ

Õ2K

è

WX´Yh·øej]Û¿ôO
h$¹$±–Õµ
CTL
ɂ
[J].
±P˜AÖ×
,2015
(7):1259.
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IF

y2



9ÔÖ×UVWXY$ˆ
52¨
ÝOh|ßñ
µmQ‚0ˆÉ¤
[J].
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,2015(1):98.
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$%«JAÚ

AC)
,2011(30):70.
[19] 
B~~

{_

{Æ¥

è

æœTݾåƤUVWXYˆ
ì./ûü¢ª$./Â.˜±òó
[J].
±P./A

,2015(1):119.
[20] 
G,

s0µ

MØQ

è

TݾÛUVWX´YômQ‚
0OhˆÉ¤
[J].
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·1821·