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浙江中医药大学 基础医学院,杭州 310053
潘晶,在读硕士,从事中医临床基础研究,E-mail:panj921@163.com
杜月光,博士,教授,硕士生导师,从事中医药对内分泌难治病研究,E-mail:duyueguang@163.com
收稿日期:2018-10-20,
网络出版日期:2018-07-12,
纸质出版日期:2019-03-20
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潘晶, 杜月光, 郭燚. 三七皂苷对TGF-
Jing PAN, Yue-guang DU, Yi GUO. Effect of Panax Notoginseng Saponins on EMT of Rat Renal Proximal Tubular Epithelial Cells Induced by TGF-β1[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(6): 89-94.
潘晶, 杜月光, 郭燚. 三七皂苷对TGF-
Jing PAN, Yue-guang DU, Yi GUO. Effect of Panax Notoginseng Saponins on EMT of Rat Renal Proximal Tubular Epithelial Cells Induced by TGF-β1[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(6): 89-94. DOI: 10.13422/j.cnki.syfjx.20190636.
目的:
2
研究三七皂苷(PNS)对转化生长因子-
β
1
(TGF-
β
1
)诱导的大鼠肾小管细胞上皮-间充质转化(EMT)的干预作用,并从沉默信息调节因子2相关酶1(SIRT1)/TGF-
β
1
/Smad通路分析其可能机制。
方法:
2
在DMEM培养基中,用10%胎牛血清培养大鼠肾小管上皮细胞(NRK-52E),分为正常组,TGF-
β
1
组(5 μg·L
-1
),白藜芦醇组(50 mg·L
-1
),SIRT1阻断剂EX527组(10 μmol·L
-1
),PNS组(100 mg·L
-1
),EX527+PNS组(EX527 10 μmol·L
-1
,PNS 100 mg·L
-1
),48 h后收集细胞;采用实时荧光定量聚合酶链式反应(Real-time PCR)检测SIRT1,
α
-平滑肌肌动蛋白(
α
-SMA),E-钙黏附蛋白(E-cadherin),TGF-
β
1
,Smad3,Smad4 mRNA表达;蛋白免疫印迹法(Western blot)检测SIRT1,
α
-SMA,E-cadherin,TGF-
β
1
蛋白表达。
结果:
2
与正常组比较,TGF-
β
1
组
α
-SMA mRNA及蛋白表达明显增多(
P
<
0.05),E-cadherin表达显著减少(
P
<
0.01);与TGF-
β
1
组比较,PNS,白藜芦醇组,E-cadherin mRNA及蛋白表达均显著增加(
P
<
0.01),
α
-SMA表达显著减少(
P
<
0.01),EMT发生被抑制,同时,SIRT1表达显著增加(
P
<
0.01),TGF-
β
1
,Smad3,Smad4表达显著降低(
P
<
0.01)。在SIRT1阻断剂EX527的干预下,PNS则不能发挥明显抑制作用。
结论:
2
PNS可以阻止TGF-
β
1
诱导的肾小管上皮细胞EMT的发生,其机制可能是通过激活SIRT1进而抑制TGF-
β
1
/Smad通路实现的。
Objective:
2
To investigate the intervention effect of panax notoginseng saponins (PNS) on epithelial-mesenchymal transition (EMT) of rat renal tubular epithelial cells (NRK-52E) induced by transforming growth factor-
β
1
(TGF-
β
1
)
and analyze the mechanism based on the silent information regulation 2 homolog 1(SIRT1)/TGF-
β
1
/Smad signaling pathway.
Method:
2
NRK-52E were cultured in DMEM medium with 10%fetal bovine serum
and divided into normal control group
TGF-
β
1
group (5 μg·L
-1
)
resveratrol (RSV) group (50 mg·L
-1
)
EX527 group (10 μmol·L
-1
)
Panax notoginseng saponins (PNS) group (100 mg·L
-1
)
and EX527+ PNS group (10 μmol·L
-1
+ 100 mg·L
-1
). Then cells were collected after drug intervention for 48 h. The expressions of
α
-SMA
E-cadherin
SIRT1
TGF-
β
1
Smad3
Smad4 mRNA in each group were detected by Real-time PCR. The protein expressions of
α
-SMA
E-cadherin
SIRT1 and TGF-
β
1
were detected by Western blot.
Result:
2
Compared with normal group
mRNA and protein expressions of
α
-SMA increased obviously(
P
<
0.05)
but E-cadherin decreased significantly(
P
<
0.01)in TGF-
β
1
group. Compared with TGF-
β
1
group
mRNA and protein expressions of
α
-SMA decreased significantly(
P
<
0.01)
while E-cadherin increased(
P
<
0.01)in resveratrol and PNS groups
and EMT was inhibited. Meanwhile
mRNA and protein expressions of SIRT1 increased significantly(
P
<
0.01)
while mRNA expressions of TGF-
β
1
Smad3
and Smad4 decreased(
P
<
0.01). Under the intervention of SIRT1 blocker EX527
PNS could not play a significant inhibitory effect on the cells.
Conclusion:
2
PNS can prevent the occurrence of EMT of renal tubular epithelial cells induced by TGF-
β
1
and the mechanism may be related to active SIRT1 to inhibit TGF-
β
1
/Smad pathway.
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