Journal of Traditional Chinese Medicine ›› 2023, Vol. 43 ›› Issue (2): 303-311.DOI: 10.19852/j.cnki.jtcm.20220922.002
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Efficacy of Qifu Lizhong enema prescription (芪附理中灌肠方) on intestinal mucosal tight junction function modulation of ulcerative colitis rat model
CHENG Jing1, Lü Shangbin2, ZHANG Yi1(
), HU Shuangyuan1, WEI Pengfei1, ZHOU Wenhua1, SHI Junfeng1
- 1 Clinical Medical college, Chengdu University of Traditional Chinese Medicine, Chengdu 610075, China
2 Basic Medical college, Chengdu University of Traditional Chinese Medicine, Chengdu 610075, China
-
Received:2022-01-11Accepted:2022-04-23Online:2023-04-15Published:2023-03-14 -
Contact:ZHANG Yi, Clinical Medical college, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China. zhangyi2288@163.com. Telephone: +86-18980880221 -
Supported by:National Natural Science Foundation of China: Research on the Biological Mechanism of Nourishing Fire and Nourishing Soil Building Muscle Regeneration in Repairing Intestinal Mucosal Barrier Damage in UC Based on MLCK/Rho/PKC-Crosstalk(81973821)
Cite this article
CHENG Jing, Lü Shangbin, ZHANG Yi, HU Shuangyuan, WEI Pengfei, ZHOU Wenhua, SHI Junfeng. Efficacy of Qifu Lizhong enema prescription (芪附理中灌肠方) on intestinal mucosal tight junction function modulation of ulcerative colitis rat model[J]. Journal of Traditional Chinese Medicine, 2023, 43(2): 303-311.
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Table 1 DAI scoring and specimen collection
| Score | Decreased body mass (%) | Stool properties | Blood in the stool |
|---|---|---|---|
| 0 | None | Normal | Occult blood test (-) |
| 1 | 1-5 | Normal | Occult blood test (-) |
| 2 | 6-10 | soft stool | Occult blood test (+) |
| 3 | 11-15 | soft stool | Occult blood test (+) |
| 4 | >15 | dilute stool | blood in the naked eye |
Table 1 DAI scoring and specimen collection
| Score | Decreased body mass (%) | Stool properties | Blood in the stool |
|---|---|---|---|
| 0 | None | Normal | Occult blood test (-) |
| 1 | 1-5 | Normal | Occult blood test (-) |
| 2 | 6-10 | soft stool | Occult blood test (+) |
| 3 | 11-15 | soft stool | Occult blood test (+) |
| 4 | >15 | dilute stool | blood in the naked eye |
Figure 1 Histopathological changes of the intestinal mucosa in various groups (hematoxylin and eosin staining, ×200 Scale bar = 100 μm, ×400 Scale bar = 200 μm). A: rat colon mucosal tissue of normal group, A1: × 200, A2: × 400 (normal group treat with physiological saline in 2 weeks); B: rat colon mucosal tissue of model group B1: × 200, B2 × 400 (model group treat with physiological saline in 2 weeks); C: rat colon mucosal tissue of mesalazine group C1: × 200, C2: × 400 (mesalazine group treat with mesalazine of 0.045 g·kg-1·d-1 in 2 weeks); D: rat colon mucosal tissue of H-QFLZ, D1: × 200, D2: × 400 (H-QFLZ treat with QFLZ of 21.6 g·kg-1·d-1 in 2 weeks); E: rat colon mucosal tissue of M-QFLZ,E1: × 200, E2: × 400 (M-QFLZ treat with QFLZ of 10.8 g·kg-1·d-1 in 2 weeks); F: rat colon mucosal tissue of L-QFLZ, F1: × 200, F2: × 400 (L-QFLZ treat with QFLZ of 5.4 g·kg-1·d-1 in 2 weeks). QFLZ: Qifulizhong enema prescription.
Figure 1 Histopathological changes of the intestinal mucosa in various groups (hematoxylin and eosin staining, ×200 Scale bar = 100 μm, ×400 Scale bar = 200 μm). A: rat colon mucosal tissue of normal group, A1: × 200, A2: × 400 (normal group treat with physiological saline in 2 weeks); B: rat colon mucosal tissue of model group B1: × 200, B2 × 400 (model group treat with physiological saline in 2 weeks); C: rat colon mucosal tissue of mesalazine group C1: × 200, C2: × 400 (mesalazine group treat with mesalazine of 0.045 g·kg-1·d-1 in 2 weeks); D: rat colon mucosal tissue of H-QFLZ, D1: × 200, D2: × 400 (H-QFLZ treat with QFLZ of 21.6 g·kg-1·d-1 in 2 weeks); E: rat colon mucosal tissue of M-QFLZ,E1: × 200, E2: × 400 (M-QFLZ treat with QFLZ of 10.8 g·kg-1·d-1 in 2 weeks); F: rat colon mucosal tissue of L-QFLZ, F1: × 200, F2: × 400 (L-QFLZ treat with QFLZ of 5.4 g·kg-1·d-1 in 2 weeks). QFLZ: Qifulizhong enema prescription.
Table 2 Effect of QFLZ on DAI scores in UC model rats
| Group | n | DAI score |
|---|---|---|
| Normal | 10 | 0 |
| Model | 10 | 2.66±0.51a |
| Mesalazine | 10 | 1.54±0.13b |
| H-QFLZ | 10 | 1.60±0.10b |
| M-QFLZ | 10 | 1.18±0.32b |
| L-QFLZ | 10 | 1.65±0.48b |
Table 2 Effect of QFLZ on DAI scores in UC model rats
| Group | n | DAI score |
|---|---|---|
| Normal | 10 | 0 |
| Model | 10 | 2.66±0.51a |
| Mesalazine | 10 | 1.54±0.13b |
| H-QFLZ | 10 | 1.60±0.10b |
| M-QFLZ | 10 | 1.18±0.32b |
| L-QFLZ | 10 | 1.65±0.48b |
Figure 2 Effects of QFLZ on the production levels of Claudin 1 and Claudin 2 in UC rats A: evaluation of Claudin 1 in rat intestinal mucosa. Claudin 1 in intestinal mucosa from various groups (IHC, ×100). A1: normal group (treated with physiological saline in 2 weeks); A2: model group (treated with physiological saline in 2 weeks); A3: mesalazine group (treated with mesalazine of 0.045 g·kg-1·d-1 in 2 weeks); A4: H-QFLZ group (treated with QFLZ of 21.6 g·kg-1·d-1 in 2 weeks); A5: M-QFLZ group (treated with QFLZ of 10.8 g·kg-1·d-1 in 2 weeks); A6: L-QFLZ group (treated with QFLZ of 5.4 g·kg-1·d-1 in 2 weeks); B: evaluation of Claudin 2 in rat intestinal mucosa. Claudin 2 in rat intestinal mucosa from various groups (IHC, ×100). B1: normal group (treated with physiological saline in 2 weeks); B2: model group (treated with physiological saline in 2 weeks); B3: mesalazine group (treated with mesalazine of 0.045 g·kg-1·d-1 in 2 weeks); B4: H-QFLZ group (treated with QFLZ of 21.6 g·kg-1·d-1 in 2 weeks); B5: medium dose QFLZ group (treated with QFLZ of 10.8 g·kg-1·d-1 in 2 weeks); B6: L-QFLZ group (treated with QFLZ of 5.4 g·kg-1·d-1 in 2 weeks); C: Claudin 1 and Claudin 2 protein expression of Western blot, 1-6 respectively represent normal group, model group, mesalazine group, H-QFLZ, M-QFLZ and L-QFLZ; E: Claudin 1 protein expression (IOD); the results are expressed as mean ± standard deviation (n = 10); F: Claudin 2 protein expression (IOD); the results are expressed as mean ± standard deviation (n = 10); G: Claudin 1 protein expression (WB); the results are expressed as mean ± standard deviation (n = 3); H: Claudin 2 protein expression (WB); the results are expressed as mean ± standard deviation (n = 3). QFLZ: Qifulizhong enema prescription; IHC: immunohistochemistry; UC: ulcerative colitis. aP < 0.05, vs normal; bP < 0.05, vs model; cP < 0.05, mesalazine.
Figure 2 Effects of QFLZ on the production levels of Claudin 1 and Claudin 2 in UC rats A: evaluation of Claudin 1 in rat intestinal mucosa. Claudin 1 in intestinal mucosa from various groups (IHC, ×100). A1: normal group (treated with physiological saline in 2 weeks); A2: model group (treated with physiological saline in 2 weeks); A3: mesalazine group (treated with mesalazine of 0.045 g·kg-1·d-1 in 2 weeks); A4: H-QFLZ group (treated with QFLZ of 21.6 g·kg-1·d-1 in 2 weeks); A5: M-QFLZ group (treated with QFLZ of 10.8 g·kg-1·d-1 in 2 weeks); A6: L-QFLZ group (treated with QFLZ of 5.4 g·kg-1·d-1 in 2 weeks); B: evaluation of Claudin 2 in rat intestinal mucosa. Claudin 2 in rat intestinal mucosa from various groups (IHC, ×100). B1: normal group (treated with physiological saline in 2 weeks); B2: model group (treated with physiological saline in 2 weeks); B3: mesalazine group (treated with mesalazine of 0.045 g·kg-1·d-1 in 2 weeks); B4: H-QFLZ group (treated with QFLZ of 21.6 g·kg-1·d-1 in 2 weeks); B5: medium dose QFLZ group (treated with QFLZ of 10.8 g·kg-1·d-1 in 2 weeks); B6: L-QFLZ group (treated with QFLZ of 5.4 g·kg-1·d-1 in 2 weeks); C: Claudin 1 and Claudin 2 protein expression of Western blot, 1-6 respectively represent normal group, model group, mesalazine group, H-QFLZ, M-QFLZ and L-QFLZ; E: Claudin 1 protein expression (IOD); the results are expressed as mean ± standard deviation (n = 10); F: Claudin 2 protein expression (IOD); the results are expressed as mean ± standard deviation (n = 10); G: Claudin 1 protein expression (WB); the results are expressed as mean ± standard deviation (n = 3); H: Claudin 2 protein expression (WB); the results are expressed as mean ± standard deviation (n = 3). QFLZ: Qifulizhong enema prescription; IHC: immunohistochemistry; UC: ulcerative colitis. aP < 0.05, vs normal; bP < 0.05, vs model; cP < 0.05, mesalazine.
Figure 3 Effects of QFLZ on the production levels of F-actin and ZO-1 in UC rats A: evaluation of F-actin in rat intestinal mucosa. F-actin in intestinal mucosa from various groups (IHC, ×100). A1: normal group (treated with physiological saline in 2 weeks); A2: model group (treated with physiological saline in 2 weeks); A3: mesalazine group (treated with mesalazine of 0.045 g·kg-1·d-1 in 2 weeks); A4: H-QFLZ group (treated with QFLZ of 21.6 g·kg-1·d-1 in 2 weeks); A5: M-QFLZ group (treated with QFLZ of 10.8 g·kg-1·d-1 in 2 weeks); A6: L-QFLZ group (treated with QFLZ of 5.4 g·kg-1·d-1 in 2 weeks); B: evaluation of ZO-1 in rat intestinal mucosa. ZO-1 in rat intestinal mucosa from various groups (IHC, ×100). B1: normal group (treated with physiological saline in 2 weeks); B2: model group (treated with physiological saline in 2 weeks); B3: mesalazine group (treated with mesalazine of 0.045 g·kg-1·d-1 in 2 weeks); B4: H-QFLZ group (treated with QFLZ of 21.6 g·kg-1·d-1 in 2 weeks); B5: medium dose QFLZ group (treated with QFLZ of 10.8 g·kg-1·d-1 in 2 weeks); B6: L-QFLZ group (treated with QFLZ of 5.4 g·kg-1·d-1 in 2 weeks); C: F-actin protein expression and ZO-1 protein expression of Western Blot, 1-6 respectively represent normal group, model group, mesalazine group, H-QFLZ, M-QFLZ and L-QFLZ; D: F-actin protein expression (IOD). The results are expressed as mean ± standard deviation (n = 10); E: ZO-1 protein expression (IOD). The results are expressed as mean ± standard deviation (n = 10); F: F-actin protein expression (WB); the results are expressed as mean ± standard deviation (n = 3); G: ZO-1 protein expression (WB); the results are expressed as mean ± standard deviation (n = 3). QFLZ: Qifulizhong enema prescription; IHC: immunohistochemistry; ZO-1: zonula occludens-1 protein; UC: ulcerative colitis. aP < 0.05, vs normal; bP < 0.05, vs model.
Figure 3 Effects of QFLZ on the production levels of F-actin and ZO-1 in UC rats A: evaluation of F-actin in rat intestinal mucosa. F-actin in intestinal mucosa from various groups (IHC, ×100). A1: normal group (treated with physiological saline in 2 weeks); A2: model group (treated with physiological saline in 2 weeks); A3: mesalazine group (treated with mesalazine of 0.045 g·kg-1·d-1 in 2 weeks); A4: H-QFLZ group (treated with QFLZ of 21.6 g·kg-1·d-1 in 2 weeks); A5: M-QFLZ group (treated with QFLZ of 10.8 g·kg-1·d-1 in 2 weeks); A6: L-QFLZ group (treated with QFLZ of 5.4 g·kg-1·d-1 in 2 weeks); B: evaluation of ZO-1 in rat intestinal mucosa. ZO-1 in rat intestinal mucosa from various groups (IHC, ×100). B1: normal group (treated with physiological saline in 2 weeks); B2: model group (treated with physiological saline in 2 weeks); B3: mesalazine group (treated with mesalazine of 0.045 g·kg-1·d-1 in 2 weeks); B4: H-QFLZ group (treated with QFLZ of 21.6 g·kg-1·d-1 in 2 weeks); B5: medium dose QFLZ group (treated with QFLZ of 10.8 g·kg-1·d-1 in 2 weeks); B6: L-QFLZ group (treated with QFLZ of 5.4 g·kg-1·d-1 in 2 weeks); C: F-actin protein expression and ZO-1 protein expression of Western Blot, 1-6 respectively represent normal group, model group, mesalazine group, H-QFLZ, M-QFLZ and L-QFLZ; D: F-actin protein expression (IOD). The results are expressed as mean ± standard deviation (n = 10); E: ZO-1 protein expression (IOD). The results are expressed as mean ± standard deviation (n = 10); F: F-actin protein expression (WB); the results are expressed as mean ± standard deviation (n = 3); G: ZO-1 protein expression (WB); the results are expressed as mean ± standard deviation (n = 3). QFLZ: Qifulizhong enema prescription; IHC: immunohistochemistry; ZO-1: zonula occludens-1 protein; UC: ulcerative colitis. aP < 0.05, vs normal; bP < 0.05, vs model.
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