Journal of Southern Medical University ›› 2026, Vol. 46 ›› Issue (3): 513-522.doi: 10.12122/j.issn.1673-4254.2026.03.05
Caixia LI1(
), Lihua CUI1, Jie CAO2, Yuxing FAN2, Xueying ZHOU3, Shukun ZHANG1, Yanjie ZUO3()
Received:2025-09-11
Online:2026-03-20
Published:2026-03-26
Contact:
Yanjie ZUO
E-mail:licaixia2013 @163.com;zyjcz9886@163.com
Caixia LI, Lihua CUI, Jie CAO, Yuxing FAN, Xueying ZHOU, Shukun ZHANG, Yanjie ZUO. Huoxue Qingjie Ling alleviates bile duct ligation-induced hepatic fibrosis in rats by regulating the Sirt1-autophagy signaling pathway[J]. Journal of Southern Medical University, 2026, 46(3): 513-522.
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URL: https://www.j-smu.com/EN/10.12122/j.issn.1673-4254.2026.03.05
| Gene | Primer sequences (5'-3') (Forward) | Primer sequences (5'-3') (Reverse) |
|---|---|---|
| FN | GATTCTTCTGGCGTCTGCAC | GATTCTTCTGGCGTCTGCAC |
| COL I | GGATAGGGACTTGTGTGA | GCTGGAAGAGTGAAGAGG |
| α-SMA | AGGGAGTGATGGTTGGAATG | GATGATGCCGTGTTCTATCG |
| Beclin-1 | AGGGAGTGATGGTTGGAATG | GATGATGCCGTGTTCTATCG |
| Atg5 | AGGGAGTGATGGTTGGAATG | GATGATGCCGTGTTCTATCG |
| LC3B | AGGGAGTGATGGTTGGAATG | GATGATGCCGTGTTCTATCG |
| P62 | ACGACTGGACGCATTTGTCT | TGGTGGGAGATGTGGGATCA |
| Sirt 1 | ACCGAGACAACCTCCTGTTG | ATGGACCTATCCGTGGCCTT |
| GAPDH | AGATGGGTGAAGGTCGGTGTG | CTGGAAGATGGTGATGGGTT |
Tab.1 Primer sequences for qRT-PCR
| Gene | Primer sequences (5'-3') (Forward) | Primer sequences (5'-3') (Reverse) |
|---|---|---|
| FN | GATTCTTCTGGCGTCTGCAC | GATTCTTCTGGCGTCTGCAC |
| COL I | GGATAGGGACTTGTGTGA | GCTGGAAGAGTGAAGAGG |
| α-SMA | AGGGAGTGATGGTTGGAATG | GATGATGCCGTGTTCTATCG |
| Beclin-1 | AGGGAGTGATGGTTGGAATG | GATGATGCCGTGTTCTATCG |
| Atg5 | AGGGAGTGATGGTTGGAATG | GATGATGCCGTGTTCTATCG |
| LC3B | AGGGAGTGATGGTTGGAATG | GATGATGCCGTGTTCTATCG |
| P62 | ACGACTGGACGCATTTGTCT | TGGTGGGAGATGTGGGATCA |
| Sirt 1 | ACCGAGACAACCTCCTGTTG | ATGGACCTATCCGTGGCCTT |
| GAPDH | AGATGGGTGAAGGTCGGTGTG | CTGGAAGATGGTGATGGGTT |
| Group | Dose (g/kg) | Body mass (g) | Liver mass (g) | Liver index (%) |
|---|---|---|---|---|
| Sham | 296.0±6.6 | 11.4±1.0 | 3.8±0.3 | |
| BDL | 198.1±11.81) | 17.9±1.81) | 9.0±0.81) | |
| HXQJL-L | 7.74 | 235.6±11.22) | 14.9±0.82) | 6.3±0.42) |
| HXQJL-H | 15.48 | 258.2±10.92)3) | 12.4±1.32)3) | 4.8±0.62) |
| EX527 | 0.005 | 163.6±8.82) | 21.4±1.02) | 13.1±1.02) |
| EX527+HXQJL | 0.005+15.48 | 204.0±13.04) | 17.5±2.14) | 8.6±0.94) |
Tab.2 Effect of Huoxue Qingjie Ling (HXQJL) on body weight, liver mass and liver index in rats with BDL-induced liver fibrosis (Mean±SD, n=8)
| Group | Dose (g/kg) | Body mass (g) | Liver mass (g) | Liver index (%) |
|---|---|---|---|---|
| Sham | 296.0±6.6 | 11.4±1.0 | 3.8±0.3 | |
| BDL | 198.1±11.81) | 17.9±1.81) | 9.0±0.81) | |
| HXQJL-L | 7.74 | 235.6±11.22) | 14.9±0.82) | 6.3±0.42) |
| HXQJL-H | 15.48 | 258.2±10.92)3) | 12.4±1.32)3) | 4.8±0.62) |
| EX527 | 0.005 | 163.6±8.82) | 21.4±1.02) | 13.1±1.02) |
| EX527+HXQJL | 0.005+15.48 | 204.0±13.04) | 17.5±2.14) | 8.6±0.94) |
| Group | Dose (g/kg) | ALT (U/L) | AST (U/L) | ALP (U/L) | GGT (U/L) | |
|---|---|---|---|---|---|---|
| Sham | 52.0±15.8 | 136.8±13.3 | 155±26.9 | 1.8±0.7 | ||
| BDL | 128.2±13.11) | 383.3±711) | 389.2±53.51) | 47.7±19.61) | ||
| HXQJL-L | 7.74 | 59.3±9.62) | 182.8±20.52) | 214.7±31.42) | 15.8±4.22) | |
| HXQJL-H | 15.48 | 53.6±7.42) | 141.9±28.12) | 179.4±57.62) | 10.1±4.92) | |
| EX527 | 0.005 | 169.7±16.42) | 490±107.82) | 577.5±70.92) | 78.2±7.72) | |
| EX527+HXQJL | 0.005+15.48 | 105.7±31.64) 289.7±101.34) | 361.2±54.24) | 41.3±14.44) | ||
Tab.3 Effect of HXQJL on serum levels of ALT, AST, ALP and GGT in rats with BDL-induced liver fibrosis (Mean±SD, n=8)
| Group | Dose (g/kg) | ALT (U/L) | AST (U/L) | ALP (U/L) | GGT (U/L) | |
|---|---|---|---|---|---|---|
| Sham | 52.0±15.8 | 136.8±13.3 | 155±26.9 | 1.8±0.7 | ||
| BDL | 128.2±13.11) | 383.3±711) | 389.2±53.51) | 47.7±19.61) | ||
| HXQJL-L | 7.74 | 59.3±9.62) | 182.8±20.52) | 214.7±31.42) | 15.8±4.22) | |
| HXQJL-H | 15.48 | 53.6±7.42) | 141.9±28.12) | 179.4±57.62) | 10.1±4.92) | |
| EX527 | 0.005 | 169.7±16.42) | 490±107.82) | 577.5±70.92) | 78.2±7.72) | |
| EX527+HXQJL | 0.005+15.48 | 105.7±31.64) 289.7±101.34) | 361.2±54.24) | 41.3±14.44) | ||
Fig.1 HE staining (Original magnification: ×200) and Sirius red staining (×100) of the liver tissue of rats in different groups.
| Group | Dose (g/kg) | Pathology score | Sirius red positive area (%) |
|---|---|---|---|
| Sham | 1.2±0.1 | 3.2±0.6 | |
| BDL | 3.9±0.31) | 47.5±7.81) | |
| HXQJL-L | 7.74 | 1.9±0.32) | 28.9±5.52) |
| HXQJL-H | 15.48 | 1.6±0.12) | 16.0±2.22) 3) |
| EX527 | 0.005 | 5.3±0.52) | 64.9±5.42) |
| EX527+HXQJL | 0.005+15.48 | 3.1±0.64) | 47.3±3.22) |
Tab.4 Effect of HXQJL on liver histopathological damage in rats with BDL-induced liver fibrosis (Mean±SD, n=8)
| Group | Dose (g/kg) | Pathology score | Sirius red positive area (%) |
|---|---|---|---|
| Sham | 1.2±0.1 | 3.2±0.6 | |
| BDL | 3.9±0.31) | 47.5±7.81) | |
| HXQJL-L | 7.74 | 1.9±0.32) | 28.9±5.52) |
| HXQJL-H | 15.48 | 1.6±0.12) | 16.0±2.22) 3) |
| EX527 | 0.005 | 5.3±0.52) | 64.9±5.42) |
| EX527+HXQJL | 0.005+15.48 | 3.1±0.64) | 47.3±3.22) |
| Group | Dose (g/kg) | FN | COL I | α-SMA |
|---|---|---|---|---|
| Sham | 1.00±0.19 | 1.00±0.08 | 1.03±0.15 | |
| BDL | 2.84±0.621) | 11.87±3.151) | 4.62±1.121) | |
| HXQJL-L | 7.74 | 1.62±0.262) | 6.07±1.192) | 2.67±0.622) |
| HXQJL-H | 15.48 | 1.36±0.442) | 3.46±0.692) | 1.79±0.472) |
| EX527 | 0.005 | 5.36±1.062) | 18.96±3.362) | 8.56±1.392) |
| EX527+HXQJL | 0.005+15.48 | 3.08±1.404) | 13.79±4.924) | 3.91±1.54) |
Tab.5 Effect of HXQJL on mRNA level of FN, COL I and α-SMAin the liver tissues of rats with BDL-induced liver fibrosis (Mean±SD, n=8)
| Group | Dose (g/kg) | FN | COL I | α-SMA |
|---|---|---|---|---|
| Sham | 1.00±0.19 | 1.00±0.08 | 1.03±0.15 | |
| BDL | 2.84±0.621) | 11.87±3.151) | 4.62±1.121) | |
| HXQJL-L | 7.74 | 1.62±0.262) | 6.07±1.192) | 2.67±0.622) |
| HXQJL-H | 15.48 | 1.36±0.442) | 3.46±0.692) | 1.79±0.472) |
| EX527 | 0.005 | 5.36±1.062) | 18.96±3.362) | 8.56±1.392) |
| EX527+HXQJL | 0.005+15.48 | 3.08±1.404) | 13.79±4.924) | 3.91±1.54) |
Fig.2 Effect of HXQJL on expression of FN, COL I and α‑SMA protein in liver tissues in rats with BDL-induced liver fibrosis. A: Sham group; B: BDL group; C: HXQJL-L group; D: HXQJL-H group; E: EX527 group; F: EX527+HXQJL group.
| Group | Dose (g/kg) | FN | COL I | α-SMA |
|---|---|---|---|---|
| Sham | 0.11±0.02 | 0.14±0.03 | 0.36±0.06 | |
| BDL | 0.36±0.021) | 0.38±0.011) | 0.75±0.071) | |
| HXQJL-L | 7.74 | 0.14±0.01 | 0.20±0.032) | 0.40±0.012) |
| HXQJL-H | 15.48 | 0.12±0.012) | 0.17±0.032) | 0.33±0.042) |
| EX527 | 0.005 | 2.33±0.172) | 1.12±0.112) | 1.22±0.102) |
| EX527+HXQJL | 0.005+15.48 | 1.44±0.134) | 0.70±0.064) | 1.00±0.024) |
Tab.6 Effect of HXQJL on protein expression levels of FN, COL I and α-SMAin the liver tissues of rats with BDL-induced liver fibrosis (Mean±SD, n=8)
| Group | Dose (g/kg) | FN | COL I | α-SMA |
|---|---|---|---|---|
| Sham | 0.11±0.02 | 0.14±0.03 | 0.36±0.06 | |
| BDL | 0.36±0.021) | 0.38±0.011) | 0.75±0.071) | |
| HXQJL-L | 7.74 | 0.14±0.01 | 0.20±0.032) | 0.40±0.012) |
| HXQJL-H | 15.48 | 0.12±0.012) | 0.17±0.032) | 0.33±0.042) |
| EX527 | 0.005 | 2.33±0.172) | 1.12±0.112) | 1.22±0.102) |
| EX527+HXQJL | 0.005+15.48 | 1.44±0.134) | 0.70±0.064) | 1.00±0.024) |
Fig.3 Effect of HXQJL onα-SMA expression in the liver tissues of rats with BDL-induced liver fibrosis(×200). A: Sham group; B: BDL group; C: HXQJL-L group; D: HXQJL-H group; E: EX527 group; F: EX527+HXQJL group.
| Groups | Dose (g/kg) | Beclin-1 | Atg5 | LC3B | p62 |
|---|---|---|---|---|---|
| Sham | 1.01±0.08 | 1.00±0.20 | 1.00±0.17 | 1.00±0.14 | |
| BDL | 2.13±0.491) | 1.61±0.121) | 1.48±0.141) | 0.63±0.101) | |
| HXQJL-L | 7.74 | 1.11±0.082) | 1.09±0.182) | 0.92±0.162) | 0.83±0.182) |
| HXQJL-H | 15.48 | 1.03±0.182) | 0.85±0.202) | 0.83±0.252) | 0.98±0.142) |
| EX527 | 0.005 | 2.99±0.252) | 2.01±1.382) | 1.92±0.172) | 0.24±0.082) |
| EX527+HXQJL | 0.005+15.48 | 1.91±0.434) | 1.5±0.304) | 1.47±0.274) | 0.68±0.164) |
Tab.7 Effect of HXQJL on mRNA levels of Beclin-1, Atg5, LC3B and p62in the liver tissues of rats with BDL-induced liver fibrosis (Mean±SD, n=8)
| Groups | Dose (g/kg) | Beclin-1 | Atg5 | LC3B | p62 |
|---|---|---|---|---|---|
| Sham | 1.01±0.08 | 1.00±0.20 | 1.00±0.17 | 1.00±0.14 | |
| BDL | 2.13±0.491) | 1.61±0.121) | 1.48±0.141) | 0.63±0.101) | |
| HXQJL-L | 7.74 | 1.11±0.082) | 1.09±0.182) | 0.92±0.162) | 0.83±0.182) |
| HXQJL-H | 15.48 | 1.03±0.182) | 0.85±0.202) | 0.83±0.252) | 0.98±0.142) |
| EX527 | 0.005 | 2.99±0.252) | 2.01±1.382) | 1.92±0.172) | 0.24±0.082) |
| EX527+HXQJL | 0.005+15.48 | 1.91±0.434) | 1.5±0.304) | 1.47±0.274) | 0.68±0.164) |
Fig. 4 Effect of HXQJL on expressions of Beclin-1, Atg5, LC3B and p62 in the liver tissues of rats with BDL-induced liver fibrosis. A: Sham group; B: BDL group; C: HXQJL-L group; D: HXQJL-H group; E: EX527 group; F: EX527+HXQJL group.
| Group | Dose (g/kg) | Beclin-1 | Atg5 | p62 | LC3B-II/LC3B-I |
|---|---|---|---|---|---|
| Sham | 0.48±0.01 | 0.60±0.04 | 1.25±0.11 | 1.81±0.01 | |
| BDL | 0.77±0.021) | 0.99±0.071) | 0.43±0.031) | 2.35±0.121) | |
| HXQJL-L | 7.74 | 0.52±0.032) | 0.73±0.032) | 0.57±0.03 | 2.14±0.022) |
| HXQJL-H | 15.48 | 0.49±0.022) | 0.67±0.042) | 0.83±0.072) | 1.83±0.022) |
| EX527 | 0.005 | 1.15±0.112) | 1.27±0.082) | 0.25±0.052) | 2.66±0.032) |
| EX527+HXQJL | 0.005+15.48 | 0.78±0.104) | 0.86±0.034) | 0.47±0.034) | 2.56±0.094) |
Tab.8 Effect of HXQJL on protein level of Beclin-1, Atg5, LC3B and p62in the liver tissues of rats with BDL-induced liver fibrosis (Mean±SD, n=8)
| Group | Dose (g/kg) | Beclin-1 | Atg5 | p62 | LC3B-II/LC3B-I |
|---|---|---|---|---|---|
| Sham | 0.48±0.01 | 0.60±0.04 | 1.25±0.11 | 1.81±0.01 | |
| BDL | 0.77±0.021) | 0.99±0.071) | 0.43±0.031) | 2.35±0.121) | |
| HXQJL-L | 7.74 | 0.52±0.032) | 0.73±0.032) | 0.57±0.03 | 2.14±0.022) |
| HXQJL-H | 15.48 | 0.49±0.022) | 0.67±0.042) | 0.83±0.072) | 1.83±0.022) |
| EX527 | 0.005 | 1.15±0.112) | 1.27±0.082) | 0.25±0.052) | 2.66±0.032) |
| EX527+HXQJL | 0.005+15.48 | 0.78±0.104) | 0.86±0.034) | 0.47±0.034) | 2.56±0.094) |
Fig.5 Effect of HXQJL on expressions of LC3B in the liver tissues of rats with BDL-induced liver fibrosis (×200). A: Sham group; B: BDL group; C: HXQJL-L group; D: HXQJL-H group; E: EX527 group; F: EX527+HXQJL group.
| Group | Dose (g/kg) | Sirt 1 |
|---|---|---|
| Sham | 1.00±0.14 | |
| BDL | 0.60±0.041) | |
| HXQJL-L | 7.74 | 0.99±0.112) |
| HXQJL-H | 15.48 | 0.99±0.082) |
| EX527 | 0.005 | 0.28±0.082) |
| EX527+HXQJL | 0.005+15.48 | 0.51±0.154) |
Tab.9 Effect of HXQJL on mRNA level ofSirt1in the liver tissues of the rats with BDL-induced liver fibrosis (Mean±SD, n=8)
| Group | Dose (g/kg) | Sirt 1 |
|---|---|---|
| Sham | 1.00±0.14 | |
| BDL | 0.60±0.041) | |
| HXQJL-L | 7.74 | 0.99±0.112) |
| HXQJL-H | 15.48 | 0.99±0.082) |
| EX527 | 0.005 | 0.28±0.082) |
| EX527+HXQJL | 0.005+15.48 | 0.51±0.154) |
Fig.6 Effect of HXQJL on expression of Sirt 1 in liver tissues in rats with BDL-induced liver fibrosis. A: Sham group; B: BDL group; C: HXQJL-L group; D: HXQJL-H group; E: EX527 group; F: EX527+HXQJL group.
| Group | Dose (g/kg) | Sirt 1 |
|---|---|---|
| Sham | 1.14±0.10 | |
| BDL | 0.43±0.041) | |
| HXQJL-L | 7.74 | 1.09±0.042) |
| HXQJL-H | 15.48 | 1.13±0.102) |
| EX527 | 0.005 | 0.21±0.012) |
| EX527+HXQJL | 0.005+15.48 | 0.39±0.034) |
Tab.10 Effect of HXQJL on protein level of Sirt1in the liver tissues of the rats with BDL-induced liver fibrosis (Mean±SD, n=8)
| Group | Dose (g/kg) | Sirt 1 |
|---|---|---|
| Sham | 1.14±0.10 | |
| BDL | 0.43±0.041) | |
| HXQJL-L | 7.74 | 1.09±0.042) |
| HXQJL-H | 15.48 | 1.13±0.102) |
| EX527 | 0.005 | 0.21±0.012) |
| EX527+HXQJL | 0.005+15.48 | 0.39±0.034) |
Fig.7 Effect of HXQJL on Sirt 1 expression in the liver tissues of the rats with BDL-induced liver fibrosis(×200). A: Sham group; B: BDL group; C: HXQJL-L group; D: HXQJL-H group; E: EX527 group; F: EX527+HXQJL group.
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