Journal of Traditional Chinese Medicine ›› 2023, Vol. 43 ›› Issue (3): 559-567.DOI: 10.19852/j.cnki.jtcm.20230208.002
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CHEN Wenlin1(), LIANG Fang1, ZHANG Yuncheng1, LV Jinzhen1, YANG Daguo2
Accepted:
2022-05-05
Online:
2023-06-15
Published:
2023-02-08
Contact:
Dr. CHEN Wenlin, Department of Liver Diseases, Shenzhen Hospital, Beijing University of Chinese Medicine, Shenzhen, Guangdong 518172, China. chenguiren69@163.com. Telephone: +86-15013727635
Supported by:
CHEN Wenlin, LIANG Fang, ZHANG Yuncheng, LV Jinzhen, YANG Daguo. Effects of Zhenggan Huayu decoction (正肝化瘀方) combined with entecavir on gut microbiota in patients with chronic hepatitis B fibrosis[J]. Journal of Traditional Chinese Medicine, 2023, 43(3): 559-567.
Before treatment (0 week) | After treatment (24 weeks) | ||||||
---|---|---|---|---|---|---|---|
Item | ETV (n = 21) | TCM (n = 38) | P value | ETV (n = 21) | TCM (n = 38) | P value | |
Sex (F/M) | 3/17 | 3/35 | 0.405 | 0.176 | 0.086 | 0.405 | |
Age | 40.45±7.35 | 41.70±9.29 | 0.456 | 40.45±7.35 | 41.70±9.29 | 0.456 | |
ALT | 86.31±105.55 | 241.84±808.09 | 0.697 | 25.32±9.48 | 26.93±12.22 | 0.563 | |
AST | 80.46±108.09 | 68.51±52.44 | 0.715 | 27.24±4.97 | 31.34±15.09 | 0.953 | |
GGT | 63.01±48.67 | 92.53±138.73 | 0.841 | 36.38±24.81 | 40.73±31.62 | 0.759 | |
ALP | 98.60±36.55 | 106.80±58.69 | 0.804 | 80.57±32.67 | 89.26±51.46 | 0.991 | |
TBA | 13.59±13.09 | 24.39±38.46 | 0.532 | 11.33±17.36 | 13.60±28.28 | 0.654 | |
TP | 73.59±6.55 | 72.82±5.75 | 0.637 | 71.79±8.05 | 74.48±5.25 | 0.328 | |
TBIL | 22.89±13.56 | 26.70±23.48 | 0.953 | 19.06±6.41 | 18.63±6.64 | 0.715 | |
DBIL | 6.01±5.98 | 8.59±12.24 | 0.741 | 3.45±1.14 | 3.97±2.80 | 0.878 | |
IBIL | 17.44±7.68 | 18.10±11.97 | 0.663 | 15.61±5.44 | 14.23±5.08 | 0.268 | |
CHE | 7485.12±2208.47 | 7555.21±2626.55 | 0.805 | 7815.45±1555.8 | 8081.00±2335.77 | 0.596 | |
TG | 1.28±0.71 | 1.37±0.69 | 0.340 | 1.16±0.41 | 1.56±1.21 | 0.396 | |
TC | 4.71±1.11 | 4.47±0.89 | 0.509 | 4.88±1.13 | 4.68±0.88 | 0.841 | |
HDL-C | 1.39±0.34 | 1.22±0.37 | 0.283 | 1.43±0.27 | 1.28±0.22 | 0.087 | |
LDL-C | 3.13±0.84 | 3.02±0.71 | 0.588 | 3.33±0.83 | 3.27±0.80 | 1.000 | |
UREA | 4.87±1.15 | 4.69±0.91 | 0.540 | 29.08±107.35 | 4.86±0.78 | 0.334 | |
CR | 72.57±13.14 | 70.72±8.75 | 0.741 | 75.46±9.86 | 73.99±11.98 | 0.841 | |
UA | 365.34±102.41 | 342.92±79.17 | 0.760 | 369.56±110.51 | 361.81±65.52 | 0.769 | |
HA | 182.92±237.93 | 202.16±272.55 | 0.800 | 87.81±88.53 | 60.15±41.98 | 0.266 | |
PIIINP | 12.13±10.30 | 11.51±9.62 | 0.888 | 8.92±5.80 | 6.63±2.14 | 0.490 | |
IV-C | 100.76±88.14 | 104.07±143.10 | 0.422 | 56.73±32.23 | 45.04±27.97 | 0.254 | |
LN | 2604.00±10286.62 | 119.10±67.09 | 0.746 | 113.61±75.35 | 93.10±38.79 | 0.526 | |
RL | 122.30±10.74 | 123.58±12.82 | 0.962 | 121.55±9.55 | 122.88±8.65 | 0.509 | |
DPV | 10.85±1.24 | 10.50±0.76 | 0.617 | 10.60±1.28 | 10.50±0.65 | 0.440 | |
TS | 41.15±5.72 | 37.38±6.42 | 0.041 | 36.55±5.01 | 35.50±5.33 | 0.594 |
Table 1 Characteristics and clinical indices of participants
Before treatment (0 week) | After treatment (24 weeks) | ||||||
---|---|---|---|---|---|---|---|
Item | ETV (n = 21) | TCM (n = 38) | P value | ETV (n = 21) | TCM (n = 38) | P value | |
Sex (F/M) | 3/17 | 3/35 | 0.405 | 0.176 | 0.086 | 0.405 | |
Age | 40.45±7.35 | 41.70±9.29 | 0.456 | 40.45±7.35 | 41.70±9.29 | 0.456 | |
ALT | 86.31±105.55 | 241.84±808.09 | 0.697 | 25.32±9.48 | 26.93±12.22 | 0.563 | |
AST | 80.46±108.09 | 68.51±52.44 | 0.715 | 27.24±4.97 | 31.34±15.09 | 0.953 | |
GGT | 63.01±48.67 | 92.53±138.73 | 0.841 | 36.38±24.81 | 40.73±31.62 | 0.759 | |
ALP | 98.60±36.55 | 106.80±58.69 | 0.804 | 80.57±32.67 | 89.26±51.46 | 0.991 | |
TBA | 13.59±13.09 | 24.39±38.46 | 0.532 | 11.33±17.36 | 13.60±28.28 | 0.654 | |
TP | 73.59±6.55 | 72.82±5.75 | 0.637 | 71.79±8.05 | 74.48±5.25 | 0.328 | |
TBIL | 22.89±13.56 | 26.70±23.48 | 0.953 | 19.06±6.41 | 18.63±6.64 | 0.715 | |
DBIL | 6.01±5.98 | 8.59±12.24 | 0.741 | 3.45±1.14 | 3.97±2.80 | 0.878 | |
IBIL | 17.44±7.68 | 18.10±11.97 | 0.663 | 15.61±5.44 | 14.23±5.08 | 0.268 | |
CHE | 7485.12±2208.47 | 7555.21±2626.55 | 0.805 | 7815.45±1555.8 | 8081.00±2335.77 | 0.596 | |
TG | 1.28±0.71 | 1.37±0.69 | 0.340 | 1.16±0.41 | 1.56±1.21 | 0.396 | |
TC | 4.71±1.11 | 4.47±0.89 | 0.509 | 4.88±1.13 | 4.68±0.88 | 0.841 | |
HDL-C | 1.39±0.34 | 1.22±0.37 | 0.283 | 1.43±0.27 | 1.28±0.22 | 0.087 | |
LDL-C | 3.13±0.84 | 3.02±0.71 | 0.588 | 3.33±0.83 | 3.27±0.80 | 1.000 | |
UREA | 4.87±1.15 | 4.69±0.91 | 0.540 | 29.08±107.35 | 4.86±0.78 | 0.334 | |
CR | 72.57±13.14 | 70.72±8.75 | 0.741 | 75.46±9.86 | 73.99±11.98 | 0.841 | |
UA | 365.34±102.41 | 342.92±79.17 | 0.760 | 369.56±110.51 | 361.81±65.52 | 0.769 | |
HA | 182.92±237.93 | 202.16±272.55 | 0.800 | 87.81±88.53 | 60.15±41.98 | 0.266 | |
PIIINP | 12.13±10.30 | 11.51±9.62 | 0.888 | 8.92±5.80 | 6.63±2.14 | 0.490 | |
IV-C | 100.76±88.14 | 104.07±143.10 | 0.422 | 56.73±32.23 | 45.04±27.97 | 0.254 | |
LN | 2604.00±10286.62 | 119.10±67.09 | 0.746 | 113.61±75.35 | 93.10±38.79 | 0.526 | |
RL | 122.30±10.74 | 123.58±12.82 | 0.962 | 121.55±9.55 | 122.88±8.65 | 0.509 | |
DPV | 10.85±1.24 | 10.50±0.76 | 0.617 | 10.60±1.28 | 10.50±0.65 | 0.440 | |
TS | 41.15±5.72 | 37.38±6.42 | 0.041 | 36.55±5.01 | 35.50±5.33 | 0.594 |
Figure 1 Effects of ETV and TCM on the gut microbiota of patients A: rarefaction curve analysis of ASVs in each group of samples. The number of identified ASVs in each group was calculated based on a randomly selected specific number of samples with 100 replacements. Quartiles numbers are plotted by a line segment; B, C: differences in gut microbiota diversity between ETV and TCM groups at different treatment periods. The significance level in the Wilcoxon signed-rank test was marked as a numerical P-value; D: the dbRDA analysis based on microbiota composition showed separations between groups. The R2 and P values generated by PERMANOVA analysis are shown in the lower-left quadrant of the plot; E: dbRDA analysis showed separations at different treatment periods within ETV and TCM groups, and R2 as well as P values generated by PERMANOVA analysis are shown at the top of the image; F: PERMANOVA analysis showed differences between treatment periods. Patients in the TCM group were treated with ZGHY 1 potion/d (oral administration in two doses) and ETV 0.5 mg/d, and the ETV group were treated with ETV 0.5 mg/d. All patients were treated for a total of 24 weeks, fecal samples were collected and analyzed in the 12 and 24 weeks, respectively. ETV: entecavir; TCM: Traditional Chinese Medicine (FZHY+ETV group); CON: control (ETV group); ZGHY: Zhenggan Huayu decoction; ASVs: amplicon sequence variants; dbRDA: distance-based redundancy analysis; PERMANOVA: permutational multivariate analysis of variance. Significant differences in TCM group: 12 w vs 24 w, aP<0.05; 0+12 w vs 24 w, aP<0.05. n = 38.
Figure 2 Microbiota composition in patients’ gut A: the composition of phyla levels in different groups. Only the top 10 members in relative abundance are shown; B: species differences at phylum levels between groups; C: the composition of family levels in different groups. Only the top 10 members in relative abundance are shown; D: species differences at family levels between groups. Patients in the TCM group were treated with ZGHY 1 potion/d (oral administration in two doses) and ETV 0.5 mg/d, and the ETV group were treated with ETV 0.5 mg/d. All patients were treated for a total of 24 weeks, fecal samples were collected and analyzed in the 12 and 24 weeks, respectively. ETV: entecavir; ZGHY: Zhenggan Huayu decoction; TCM: Traditional Chinese Medicine (FZHY + ETV group); CON: control (ETV group).
Figure 3 Heat maps show correlations between feature genera and clinical indicators Spearman correlation test showed that: * q<0.05; + q<0.01. A correlation coefficient>0.6 was considered as a positive correlation, and<-0.6 as a negative correlation. ALP: alkaline phosphatase; ALT: alanine aminotransferase; AST: aspartate aminotransferase; CHE: cholinesterase; CR: creatinine; DBIL: direct bilirubin; GGT: γ-glutamyl transferase; HA: hyaluronic acid; HDL-C: high-density lipoprotein cholesterol; IBIL: indirect bilirubin; IV-C: Type IV collagen; LDL-C: low-density lipoprotein cholesterol; LN: laminin; PIIINP: type III procollagen peptide; TBA: total bile acid; TBIL: total bilirubin; TC: total cholesterol; TG: triglyceride; TP: total protein; UA: uric acid; UREA: urea; RL: right lobe of liver oblique diameter; DPV: portal vein diameter; TS: spleen thickness.
Figure 4 Effects of ETV and TCM on gut microbiota functions A: rarefaction curve analysis of KOs in each group of samples. The number of annotated KO genes in each group was calculated based on a randomly selected specific number of samples with 100 replacements. Quartile numbers were plotted by a line segment. B: boxplot showing the Shannon index of gut microbiota function for patients in ETV and TCM groups at different treatment periods, revealing microbiota function diversity in patients’ gut. C: fold change of gut microbiota function between patients in TCM and ETV groups. Only functional items with significant differences (P<0.05) are shown, TCM group: n = 38, ETV group, n = 21. Patients in the TCM group were treated with ZGHY 1 potion/d (oral administration in two doses) and ETV 0.5 mg/d, and the ETV group were treated with ETV 0.5 mg/d. All patients were treated for a total of 24 weeks, fecal samples were collected and analyzed in the 12 and 24 weeks, respectively. ETV: entecavir; TCM: Traditional Chinese Medicine (FZHY + ETV group); CON: control (ETV group); KO: Kyoto Encyclopedia of Genes and Genomes Orthology; ZGHY: Zhenggan Huayu decoction.
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