By studying the effect of Buzhong Yiqi Decoction(补中益气汤)on the endoplasmic reticulum stress of A549 cells and A549/DDP cells through Nrf2/ROSpathway,the molecular mechanism of Buzhong Yiqi Decoction improving cisplatin resistance in non-small cell lung cancer was investigated.
The serum containing Buzhong Yiqi Decoction was prepared and A549 cells and A549/DDP cells were cultured. They were randomly divided into group A(A549 cells),Group B(A549 cells+20μg/mL cisplatin),group C(A549 cells+20μg/mL cisplatin+10% Buzhong Yiqi Decoction-containing serum),group D(A549/DDP cells),group E(A549/DDP cells+20μg/mL cisplatin)and group F(A549/DDP cells+20μg/mL cisplatin+10% Buzhong Yiqi Decoction-containing serum). Half inhibitory concentration of cisplatin(IC50)was detected by CCK-8 method,and reactive oxygen species(ROS)were detected by DCFH-DA fluorescent probe method. The protein expression levels of Nrf2,P-Nrf2,PERK,P-PERK,eIF2α,P-eIF2α,IRE1 and XBP1 were detected by Western blot.
In both A549 cells and A549/DDP cells,Buzhong Yiqi Decoction significantly decreased the IC50 value of cisplatin(P<0.05). In both cells,cisplatin could up-regulate the protein expressions of Nrf2 and P-Nrf2,and after the intervention of Buzhong Yiqi Decoction,the expression levels of both were down-regulated(P<0.05)in A549/DDPcells compared with those in A549 cells(P<0.05). Compared with the blank group,cisplatin could up-regulate the ROSexpression of the two cells(P<0.05),the ROS expression was further up-regulated after combined with Buzhong Yiqi Decoction(P<0.05),and the ROSexpression level of A549 cells was significantly higher than that of A549/DDP cells(P<0.05). There were no significant differences in the expressions of PERK and eIF2αamong all groups(P>0.05). Cisplatin significantly up-regulated the protein expressions of PPERK,P-eIF2α,IRE1 and XBP1 in the two cell lines,and the expression was up-regulated again after the intervention of Buzhong Yiqi Decoction(P<0.05),and the expression of ER stress protein in A549 cells was higher than that in A549/DDP cells(P<0.05).
Buzhong Yiqi Decoction may improve cisplatin resistance of A549 cells and A549/DDP cells by regulating ERSthrough Nrf2/ROSsignal pathway.
| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |