Tianjin Medical Journal ›› 2023, Vol. 51 ›› Issue (4): 360-365.doi: 10.11958/20221110
• Experimental Research • Previous Articles Next Articles
TANG Qiaofei1(), CAO Jianqiu2, ZHANG Shuang1,△(
)
Received:
2022-07-14
Revised:
2022-09-22
Published:
2023-04-15
Online:
2023-04-20
Contact:
ZHANG Shuang
E-mail:tanghuashengmi@163.com;zs_5257@163.com
TANG Qiaofei, CAO Jianqiu, ZHANG Shuang. Study on the mechanism of let-7e-5p regulating Fas/FasL to affect the pathogenesis of allergic rhinitis in mice[J]. Tianjin Medical Journal, 2023, 51(4): 360-365.
CLC Number:
分级评 分(分) | 喷嚏 (个/30 min) | 清涕 | 搔鼻(次) |
---|---|---|---|
0 | 0 | 无 | 无 |
1 | 1~3 | 流涕至鼻前孔 | 轻微抓鼻几次 |
2 | 4~10 | 流涕超过鼻前孔 | 双爪反复挠鼻 |
3 | >11 | 涕流满面 | 抓挠鼻面不止、四处摩擦 |
Tab.1 Score criteria for allergic rhinitis in mice
分级评 分(分) | 喷嚏 (个/30 min) | 清涕 | 搔鼻(次) |
---|---|---|---|
0 | 0 | 无 | 无 |
1 | 1~3 | 流涕至鼻前孔 | 轻微抓鼻几次 |
2 | 4~10 | 流涕超过鼻前孔 | 双爪反复挠鼻 |
3 | >11 | 涕流满面 | 抓挠鼻面不止、四处摩擦 |
基因名称 | 引物序列(5'→3') | 产物大小(bp) |
---|---|---|
let-7e-5p | 上游:TGAGGTAGGAGGTTGTATAGTT | 54 |
下游:GCAGGGTCCGAGGTATTC | ||
U6 | 上游:CGCAAGGATGACACGCAAAT | 74 |
下游:GCAGGGTCCGAGGTATTC | ||
Fas | 上游:AACCAGACTTCTACTGCGATTC | 144 |
下游:TCAACAACCATAGGCGATTT | ||
FasL | 上游:GCCATCACAACCACTCCC | 107 |
下游:CCAGAGCCACCAGAACCA | ||
β-actin | 上游:CTGTGCCCATCTACGAGGGCTAT | 155 |
下游:TTTGATGTCACGCACGATTTCC |
Tab.2 The primer sequence for real-time PCR
基因名称 | 引物序列(5'→3') | 产物大小(bp) |
---|---|---|
let-7e-5p | 上游:TGAGGTAGGAGGTTGTATAGTT | 54 |
下游:GCAGGGTCCGAGGTATTC | ||
U6 | 上游:CGCAAGGATGACACGCAAAT | 74 |
下游:GCAGGGTCCGAGGTATTC | ||
Fas | 上游:AACCAGACTTCTACTGCGATTC | 144 |
下游:TCAACAACCATAGGCGATTT | ||
FasL | 上游:GCCATCACAACCACTCCC | 107 |
下游:CCAGAGCCACCAGAACCA | ||
β-actin | 上游:CTGTGCCCATCTACGAGGGCTAT | 155 |
下游:TTTGATGTCACGCACGATTTCC |
组别 | 评分(分) | 嗜酸性粒细胞(个/视野) |
---|---|---|
Control组 | 1.33±0.58 | 21.50±2.65 |
AR组 | 7.67±0.58a | 147.75±6.75a |
AR+agomir-NC组 | 7.33±0.58a | 136.25±6.13a |
AR+agomir组 | 4.00±1.00b | 75.00±10.42b |
AR+antagomir-NC组 | 7.33±0.58a | 149.75±1.71a |
AR+antagomir组 | 9.00±0.00c | 226.25±8.18c |
F | 63.310** | 441.500** |
Tab.3 Comparison of allergic rhinitis score and eosinophil number between the six groups of mice
组别 | 评分(分) | 嗜酸性粒细胞(个/视野) |
---|---|---|
Control组 | 1.33±0.58 | 21.50±2.65 |
AR组 | 7.67±0.58a | 147.75±6.75a |
AR+agomir-NC组 | 7.33±0.58a | 136.25±6.13a |
AR+agomir组 | 4.00±1.00b | 75.00±10.42b |
AR+antagomir-NC组 | 7.33±0.58a | 149.75±1.71a |
AR+antagomir组 | 9.00±0.00c | 226.25±8.18c |
F | 63.310** | 441.500** |
组别 | let-7e-5p | Fas | FasL |
---|---|---|---|
Control组 | 1.00±0.07 | 1.00±0.07 | 1.00±0.06 |
AR组 | 0.30±0.00a | 3.53±0.16a | 2.80±0.05a |
AR+agomir-NC组 | 0.34±0.02a | 3.34±0.25a | 2.41±0.04a |
AR+agomir组 | 0.70±0.04b | 1.98±0.05b | 1.33±0.13b |
AR+antagomir-NC组 | 0.31±0.02a | 3.48±0.36a | 2.59±0.16a |
AR+antagomir组 | 0.20±0.00c | 4.59±0.30c | 3.59±0.22c |
F | 226.200** | 92.260** | 173.300** |
Tab.4 Comparison of mRNA expression of let-7e-5p,Fas and FasL in nasal mucosa between the six groups of mice
组别 | let-7e-5p | Fas | FasL |
---|---|---|---|
Control组 | 1.00±0.07 | 1.00±0.07 | 1.00±0.06 |
AR组 | 0.30±0.00a | 3.53±0.16a | 2.80±0.05a |
AR+agomir-NC组 | 0.34±0.02a | 3.34±0.25a | 2.41±0.04a |
AR+agomir组 | 0.70±0.04b | 1.98±0.05b | 1.33±0.13b |
AR+antagomir-NC组 | 0.31±0.02a | 3.48±0.36a | 2.59±0.16a |
AR+antagomir组 | 0.20±0.00c | 4.59±0.30c | 3.59±0.22c |
F | 226.200** | 92.260** | 173.300** |
组别 | Fas | FasL |
---|---|---|
Control组 | 1.00±0.02 | 1.01±0.03 |
AR组 | 4.91±0.12a | 3.15±0.06a |
AR+agomir-NC组 | 4.85±0.07a | 3.12±0.09a |
AR+agomir组 | 1.97±0.09b | 1.82±0.15b |
AR+antagomir-NC组 | 4.64±0.14a | 3.14±0.10a |
AR+antagomir组 | 5.57±0.37c | 4.50±0.16c |
F | 334.900** | 382.100** |
Tab.5 Comparison of protein expression levels of Fas and FasL in nasal mucosa between the six groups of mice
组别 | Fas | FasL |
---|---|---|
Control组 | 1.00±0.02 | 1.01±0.03 |
AR组 | 4.91±0.12a | 3.15±0.06a |
AR+agomir-NC组 | 4.85±0.07a | 3.12±0.09a |
AR+agomir组 | 1.97±0.09b | 1.82±0.15b |
AR+antagomir-NC组 | 4.64±0.14a | 3.14±0.10a |
AR+antagomir组 | 5.57±0.37c | 4.50±0.16c |
F | 334.900** | 382.100** |
组别 | IgE (μg/L) | sIgE (μg/L) | 1型辅助性T细胞(Th1) | 2型辅助性T细胞(Th2) | ||
---|---|---|---|---|---|---|
IL-12(ng/L) | IFN-γ(ng/L) | IL-4(ng/L) | IL-13(ng/L) | |||
Control组 | 28.77±3.15 | 2.18±0.24 | 186.44±20.53 | 70.77±8.24 | 29.14±2.93 | 18.56±1.97 |
AR组 | 179.68±16.82a | 39.37±3.98a | 93.57±10.48a | 32.62±3.73a | 72.93±7.49a | 146.29±16.68a |
AR+agomir-NC组 | 192.71±21.11a | 35.67±3.59a | 89.58±9.79a | 34.47±4.19a | 73.87±8.21a | 151.06±17.66a |
AR+agomir组 | 48.16±4.14ab | 26.56±3.11ab | 167.82±18.15b | 55.21±6.78ab | 49.96±5.67ab | 106.80±10.84ab |
AR+antagomir-NC组 | 187.66±21.23a | 39.63±4.00a | 91.68±10.14a | 32.71±3.83a | 74.71±8.20a | 150.27±16.83a |
AR+antagomir组 | 272.61±28.86abc | 55.23±6.35abc | 75.08±8.56ab | 20.77±2.39ab | 116.89±14.16abc | 195.96±20.68abc |
F | 78.040** | 59.920** | 35.280** | 36.440** | 35.960** | 46.590** |
Tab.6 Changes of serum IgE,sIgE,IL-12,IFN-γ,IL-4 and IL-13 levels in each group
组别 | IgE (μg/L) | sIgE (μg/L) | 1型辅助性T细胞(Th1) | 2型辅助性T细胞(Th2) | ||
---|---|---|---|---|---|---|
IL-12(ng/L) | IFN-γ(ng/L) | IL-4(ng/L) | IL-13(ng/L) | |||
Control组 | 28.77±3.15 | 2.18±0.24 | 186.44±20.53 | 70.77±8.24 | 29.14±2.93 | 18.56±1.97 |
AR组 | 179.68±16.82a | 39.37±3.98a | 93.57±10.48a | 32.62±3.73a | 72.93±7.49a | 146.29±16.68a |
AR+agomir-NC组 | 192.71±21.11a | 35.67±3.59a | 89.58±9.79a | 34.47±4.19a | 73.87±8.21a | 151.06±17.66a |
AR+agomir组 | 48.16±4.14ab | 26.56±3.11ab | 167.82±18.15b | 55.21±6.78ab | 49.96±5.67ab | 106.80±10.84ab |
AR+antagomir-NC组 | 187.66±21.23a | 39.63±4.00a | 91.68±10.14a | 32.71±3.83a | 74.71±8.20a | 150.27±16.83a |
AR+antagomir组 | 272.61±28.86abc | 55.23±6.35abc | 75.08±8.56ab | 20.77±2.39ab | 116.89±14.16abc | 195.96±20.68abc |
F | 78.040** | 59.920** | 35.280** | 36.440** | 35.960** | 46.590** |
组别 | Fas | FasL |
---|---|---|
NC+3'UTR-WT组 | 0.92±0.12 | 0.98±0.15 |
let-7e-5p agomir+3'UTR-WT组 | 0.49±0.06a | 0.50±0.06a |
NC+3'UTR-MT组 | 1.00±0.11 | 1.00±0.13 |
let-7e-5p agomir+3'UTR-MT组 | 0.91±0.13b | 1.02±0.11b |
F | 13.230** | 13.740** |
Tab.7 Comparison of luciferase relative activity between the four groups
组别 | Fas | FasL |
---|---|---|
NC+3'UTR-WT组 | 0.92±0.12 | 0.98±0.15 |
let-7e-5p agomir+3'UTR-WT组 | 0.49±0.06a | 0.50±0.06a |
NC+3'UTR-MT组 | 1.00±0.11 | 1.00±0.13 |
let-7e-5p agomir+3'UTR-MT组 | 0.91±0.13b | 1.02±0.11b |
F | 13.230** | 13.740** |
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