Journal of Traditional Chinese Medicine ›› 2023, Vol. 43 ›› Issue (5): 963-972.DOI: 10.19852/j.cnki.jtcm.20230608.001
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Electroacupuncture stimulating Zhongji (CV3), Guanyuan (CV4), and bilateral Dahe (KI12) attenuates inflammation in rats with chronic nonbacterial prostatitis induced by estradiol through inhibiting toll-like receptor 4 pathway
LI Zhihao1, HAN Wenjun3, SONG Xiuling4, LI Yan1(
), CHEN Yuelai2()
- 1 Department of Acupuncture-Moxibustion, Tuina and Traumatology, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiaotong University Shool of Medicine, Shanghai 200233, China
2 Sleep Medical Center, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
3 Department of Andrology, the Seventh People's Hospital affiliated to Shanghai University of Traditional Chinese Medicine, Shanghai 200137, China
4 Graduate School of Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
-
Received:2022-05-22Accepted:2022-08-15Online:2023-10-15Published:2023-06-08 -
Contact:CHEN Yuelai, Sleep Medical Center, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China.chenyuelai@163.com ; LI Yan, Department of Acupuncture-Moxibustion, Tuina and Traumatology, Shanghai Sixth People's Hospital Affiliated to Shanghai Jiaotong University Shool of Medicine, Shanghai 200233, China.0001liyan@163.com . Telephone: +86-21-51322519 -
Supported by:Program of Shanghai Pudong New Area Famous Traditional Chinese Medicine(PWRzm2020-01)
Cite this article
LI Zhihao, HAN Wenjun, SONG Xiuling, LI Yan, CHEN Yuelai. Electroacupuncture stimulating Zhongji (CV3), Guanyuan (CV4), and bilateral Dahe (KI12) attenuates inflammation in rats with chronic nonbacterial prostatitis induced by estradiol through inhibiting toll-like receptor 4 pathway[J]. Journal of Traditional Chinese Medicine, 2023, 43(5): 963-972.
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Table 1 Prostate weight and prostate index of CNP rats in each group ($\bar{x}±s$)
| Group | n | Prostate weight (g) | Prostate index |
|---|---|---|---|
| Control | 8 | 1.14±0.10 | 2.36±0.22 |
| Model | 8 | 0.61±0.07a | 1.71±0.28a |
| EA | 8 | 0.45±0.07b | 1.28±0.32c |
Table 1 Prostate weight and prostate index of CNP rats in each group ($\bar{x}±s$)
| Group | n | Prostate weight (g) | Prostate index |
|---|---|---|---|
| Control | 8 | 1.14±0.10 | 2.36±0.22 |
| Model | 8 | 0.61±0.07a | 1.71±0.28a |
| EA | 8 | 0.45±0.07b | 1.28±0.32c |
Figure 1 Effects of EA on mechanical withdrawal threshold in CNP rats Control group: treated with sham operation and hydrogenated soybean oil; model group: treated with surgical castration and 17-β estradiol; EA group: treated with surgical castration, 17-β estradiol and EA for eight days. CNP: chronic nonbacterial prostatitis; EA: electroacupuncture. Data are shown as the mean ± standard deviation (repeated-measures analysis of variance and Sidak test), n = 8 rats/group. aP < 0.05 compared with the control group; bP < 0.01 compared with the control group; cP < 0.05 compared with the model group; dP < 0.01 compared with the model group.
Figure 1 Effects of EA on mechanical withdrawal threshold in CNP rats Control group: treated with sham operation and hydrogenated soybean oil; model group: treated with surgical castration and 17-β estradiol; EA group: treated with surgical castration, 17-β estradiol and EA for eight days. CNP: chronic nonbacterial prostatitis; EA: electroacupuncture. Data are shown as the mean ± standard deviation (repeated-measures analysis of variance and Sidak test), n = 8 rats/group. aP < 0.05 compared with the control group; bP < 0.01 compared with the control group; cP < 0.05 compared with the model group; dP < 0.01 compared with the model group.
Figure 2 Representative histological images of prostates from each group A-C: Prostate sections from all groups were stained with HE for morphological changes (× 100). A1-C1: selection part of images of A-C (× 200). A, A1: Control group; B, B1: Model group; C, C1: EA group. Control group: treated with sham operation and hydrogenated soybean oil; model group: treated with surgical castration and 17-β estradiol; EA group: treated with surgical castration, 17-β estradiol and EA for eight days. EA: electroacupuncture. The red arrow points to the blood vessel (normal structure); the black arrow points to the infiltration of inflammatory cells; the blue arrow points to interstitial edema; and the green arrow points to congestion in the prostates.
Figure 2 Representative histological images of prostates from each group A-C: Prostate sections from all groups were stained with HE for morphological changes (× 100). A1-C1: selection part of images of A-C (× 200). A, A1: Control group; B, B1: Model group; C, C1: EA group. Control group: treated with sham operation and hydrogenated soybean oil; model group: treated with surgical castration and 17-β estradiol; EA group: treated with surgical castration, 17-β estradiol and EA for eight days. EA: electroacupuncture. The red arrow points to the blood vessel (normal structure); the black arrow points to the infiltration of inflammatory cells; the blue arrow points to interstitial edema; and the green arrow points to congestion in the prostates.
Table 2 Level of inflammatory factors in the prostates (pg/mL, $\bar{x}±s$)
| Group | n | IL-1β | IL-6 | TNF-α |
|---|---|---|---|---|
| Control | 8 | 253.4±25.5 | 15.7±7.5 | 6.6±1.2 |
| Model | 8 | 504.2±29.3a | 64.9±5.6a | 34.8±8.6a |
| EA | 8 | 396.6±28.8b | 33.8±3.6b | 19.7±3.6c |
Table 2 Level of inflammatory factors in the prostates (pg/mL, $\bar{x}±s$)
| Group | n | IL-1β | IL-6 | TNF-α |
|---|---|---|---|---|
| Control | 8 | 253.4±25.5 | 15.7±7.5 | 6.6±1.2 |
| Model | 8 | 504.2±29.3a | 64.9±5.6a | 34.8±8.6a |
| EA | 8 | 396.6±28.8b | 33.8±3.6b | 19.7±3.6c |
Figure 3 TLR4, MyD88, IκB-α, and NF-κB expression in prostates from each group A: Western blotting bands for TLR4; B: Western blotting bands for MyD88; C: Western blotting bands for IκB-α and p-IκBα; D: Western blotting bands for NF-κB p65 and p-p65. Control: treated with sham operation and hydrogenated soybean oil; model: treated with surgical castration and 17-β estradiol; EA: treated with surgical castration, 17-β estradiol and EA for eight days. EA: electroacupuncture; TLR4: toll-like receptor 4; MyD88: myeloid differentiation factor 88; IκBα: inhibitors of kappa‐B alpha; NF-κB: nuclear factor-kappa B. Data are shown as the mean ± standard deviation (one-way analysis of variance and post hoc Tukey’s test), n = 8 rats/group. aP < 0.01, compared with the control group; bP < 0.05, compared with the model group.
Figure 3 TLR4, MyD88, IκB-α, and NF-κB expression in prostates from each group A: Western blotting bands for TLR4; B: Western blotting bands for MyD88; C: Western blotting bands for IκB-α and p-IκBα; D: Western blotting bands for NF-κB p65 and p-p65. Control: treated with sham operation and hydrogenated soybean oil; model: treated with surgical castration and 17-β estradiol; EA: treated with surgical castration, 17-β estradiol and EA for eight days. EA: electroacupuncture; TLR4: toll-like receptor 4; MyD88: myeloid differentiation factor 88; IκBα: inhibitors of kappa‐B alpha; NF-κB: nuclear factor-kappa B. Data are shown as the mean ± standard deviation (one-way analysis of variance and post hoc Tukey’s test), n = 8 rats/group. aP < 0.01, compared with the control group; bP < 0.05, compared with the model group.
Figure 4 Level of TLR4 expression and nuclear translocation of NF-κB A: prostate sections were stained with immunofluorescent staining of TLR4 (× 200). A1-A3: control group, A4-A6: model group, A7-A9: EA group. A1, A4, A7: nuclei were stained with DAPI in blue; A2, A5. A8: TLR4 were stained in red; A3, A6, A9: immunofluorescent staining of TLR4 (red) with the DAPI (blue). B: the bar graph shows the quantitative analysis of the mean fluorescence intensity of TLR4. aP < 0.01 compared with the control group; bP < 0.05, compared with the model group. C: prostate sections were stained with immunofluorescent staining of NF-κB (× 200). C1-C4: control group, C5-C8: model group, C9-C12: EA group. C1, C5, C9: nuclei were stained with DAPI in blue; C2, C6. C10: p65 were stained in green; C3, C7, C11: immunofluorescent staining of NF-κB p65 (green) with the DAPI (blue); C4, C8. C12: Selection part of images of C3, C7, C11 (× 400). The arrow indicates the nuclear translocation of NF-κB p65. EA: electroacupuncture; TLR4: toll-like receptor 4; DAPI: 4',6-Diamidino-2-Phenylindole, Dihydrochloride; NF-κB: nuclear factor-kappa B. Data are shown as the mean ± standard deviation (one-way analysis of variance and post hoc Tukey’s test), n = 8 rats/group.
Figure 4 Level of TLR4 expression and nuclear translocation of NF-κB A: prostate sections were stained with immunofluorescent staining of TLR4 (× 200). A1-A3: control group, A4-A6: model group, A7-A9: EA group. A1, A4, A7: nuclei were stained with DAPI in blue; A2, A5. A8: TLR4 were stained in red; A3, A6, A9: immunofluorescent staining of TLR4 (red) with the DAPI (blue). B: the bar graph shows the quantitative analysis of the mean fluorescence intensity of TLR4. aP < 0.01 compared with the control group; bP < 0.05, compared with the model group. C: prostate sections were stained with immunofluorescent staining of NF-κB (× 200). C1-C4: control group, C5-C8: model group, C9-C12: EA group. C1, C5, C9: nuclei were stained with DAPI in blue; C2, C6. C10: p65 were stained in green; C3, C7, C11: immunofluorescent staining of NF-κB p65 (green) with the DAPI (blue); C4, C8. C12: Selection part of images of C3, C7, C11 (× 400). The arrow indicates the nuclear translocation of NF-κB p65. EA: electroacupuncture; TLR4: toll-like receptor 4; DAPI: 4',6-Diamidino-2-Phenylindole, Dihydrochloride; NF-κB: nuclear factor-kappa B. Data are shown as the mean ± standard deviation (one-way analysis of variance and post hoc Tukey’s test), n = 8 rats/group.
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