Journal of Traditional Chinese Medicine ›› 2024, Vol. 44 ›› Issue (4): 652-659.DOI: 10.19852/j.cnki.jtcm.2024.04.003
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Ganoderma Lucidum Spore Oil enhances the effect of cyclophosphamide via inhibiting programmed death-1 and prolongs the survival of H22 tumor-bearing mice
JIANG Zhaojian1, CAI Hongfei1, YUAN Cheng1, CAO Lin1, XU Wendong1, HAN Yaming1, ZHANG Qin1, LI Jing1, WANG Qin2(
), LIU Juyan3,4()
- 1 Guangzhou HanFang Pharmaceutical Company Limited, National Engineering Research Center of Pharmaceutical Processing Technology of Traditional Chinese Medicine and Drug Innovation, Guangdong Provincial Key Laboratory of Medicinal Lipids, Guangzhou 510240, China
2 Zhongshan Medical College, Sun Yat-sen University, Guangzhou 510080, China
3 Guangzhou Pharmaceutical Holdings Limited, Guangzhou 510130, China
4 National Engineering Research Center of Pharmaceutical Processing Technology of Traditional Chinese Medicine and Drug Innovation, Guangzhou 510240, China
-
Received:2023-02-16Accepted:2023-07-20Online:2024-08-15Published:2024-07-15 -
Contact:WANG Qin, Zhongshan Medical College, Sun Yat-sen University, Guangzhou 510080, China.wangqin6@mail.sysu.edu.cn ; LIU Juyan, Guangzhou Pharmaceutical Holdings Limited, Guangzhou 510130, China.LJYan65@163.com Telephone: + 86-13710583307 -
Supported by:Ministry of Science and Technology of China: Evaluation, Re-examination, Promotion and Demonstration of Complete Sets of Technical Equipment for Efficient and Energy-Saving Extraction and Separation of Traditional Chinese Medicine(2017YFC1703104);Key Laboratory Project of Pharmaceutical Lipids in Guangdong Province : Assessment of Enterprise-Class Key Laboratory of Guangdong Province(2020B1212070024);Guangdong Province Key Areas Research and Development Program Project : Research and Development on Innovative Key Technologies and Intelligent Production Equipment about Processing, Decoction, Extraction and Separation of Traditional Chinese Medicine(2020B1111120002)
Cite this article
JIANG Zhaojian, CAI Hongfei, YUAN Cheng, CAO Lin, XU Wendong, HAN Yaming, ZHANG Qin, LI Jing, WANG Qin, LIU Juyan. Ganoderma Lucidum Spore Oil enhances the effect of cyclophosphamide via inhibiting programmed death-1 and prolongs the survival of H22 tumor-bearing mice[J]. Journal of Traditional Chinese Medicine, 2024, 44(4): 652-659.
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Figure 1 Effect of GLSO and CTX on tumor growth and body weight of H22 tumor-bearing mice in different groups A: size of tumors in each group; B: weight of tumors in each group; C: body weight of mice in each group. Control group: mice were given normal saline; GLSO group: mice were given 4.68 g/kg GLSO and normal saline; CTX group: mice were given normal saline and 50 mg/kg CTX; Vehicle + CTX: mice were given vehicle and 50 mg/kg CTX; GLSO-H + CTX: mice were given 4.68 g/kg GLSO and 50 mg/kg CTX; GLSO-M + CTX: mice were given 2.496 g/kg GLSO and 50 mg/kg CTX; GLSO-L + CTX: mice were given 0.312 g/kg GLSO and 50 mg/kg CTX. GLSO: Ganoderma Lucidum Spore Oil; CTX: cyclophosphamide. Data are presented as the mean ± standard deviation using one-way analysis of variance (n = 6-8). aP < 0.001 vs Control group; bP < 0.001, cP < 0.01 vs Vehicle + CTX group.
Figure 1 Effect of GLSO and CTX on tumor growth and body weight of H22 tumor-bearing mice in different groups A: size of tumors in each group; B: weight of tumors in each group; C: body weight of mice in each group. Control group: mice were given normal saline; GLSO group: mice were given 4.68 g/kg GLSO and normal saline; CTX group: mice were given normal saline and 50 mg/kg CTX; Vehicle + CTX: mice were given vehicle and 50 mg/kg CTX; GLSO-H + CTX: mice were given 4.68 g/kg GLSO and 50 mg/kg CTX; GLSO-M + CTX: mice were given 2.496 g/kg GLSO and 50 mg/kg CTX; GLSO-L + CTX: mice were given 0.312 g/kg GLSO and 50 mg/kg CTX. GLSO: Ganoderma Lucidum Spore Oil; CTX: cyclophosphamide. Data are presented as the mean ± standard deviation using one-way analysis of variance (n = 6-8). aP < 0.001 vs Control group; bP < 0.001, cP < 0.01 vs Vehicle + CTX group.
Table 1 Survival of mice in each group ($\bar{x}±s$)
| Group | n | Average survival time (days) | Increased life span (%) | Median survival time (days) |
|---|---|---|---|---|
| Control | 24 | 32.5±2.7a | - | 28.0±3.1a |
| GLSO | 22 | 35.9±3.4 | 10.23 | 33.0±2.4 |
| Vehicle+CTX | 25 | 41.1±2.8 | 26.37 | 45.0±4.0 |
| GLSO-H+CTX | 24 | 44.6±3.9 | 37.15 | 49.0±12.3 |
| GLSO-M+CTX | 25 | 54.0±3.8b | 65.83 | 62.0±5.8b |
| GLSO-L+CTX | 25 | 54.7±3.9a | 68.04 | 62.0±10.8a |
Table 1 Survival of mice in each group ($\bar{x}±s$)
| Group | n | Average survival time (days) | Increased life span (%) | Median survival time (days) |
|---|---|---|---|---|
| Control | 24 | 32.5±2.7a | - | 28.0±3.1a |
| GLSO | 22 | 35.9±3.4 | 10.23 | 33.0±2.4 |
| Vehicle+CTX | 25 | 41.1±2.8 | 26.37 | 45.0±4.0 |
| GLSO-H+CTX | 24 | 44.6±3.9 | 37.15 | 49.0±12.3 |
| GLSO-M+CTX | 25 | 54.0±3.8b | 65.83 | 62.0±5.8b |
| GLSO-L+CTX | 25 | 54.7±3.9a | 68.04 | 62.0±10.8a |
Figure 2 Effect of GLSO on the survival of H22 tumor-bearing mice treated with CTX in different groups Control group: mice were given normal saline; GLSO group: mice were given 4.68 g/kg GLSO and normal saline; Vehicle + CTX: mice were given vehicle and 50 mg/kg CTX; GLSO-H + CTX: mice were given 4.68 g/kg GLSO and 50 mg/kg CTX; GLSO-M + CTX: mice were given 2.496 g/kg GLSO and 50 mg/kg CTX; GLSO-L + CTX: mice were given 0.312 g/kg GLSO and 50 mg/kg CTX. GLSO: Ganoderma Lucidum Spore Oil; CTX: cyclophos-phamide. Kaplan-Meier survival analysis was employed to compare the difference of survival rates between groups (n = 22-25).
Figure 2 Effect of GLSO on the survival of H22 tumor-bearing mice treated with CTX in different groups Control group: mice were given normal saline; GLSO group: mice were given 4.68 g/kg GLSO and normal saline; Vehicle + CTX: mice were given vehicle and 50 mg/kg CTX; GLSO-H + CTX: mice were given 4.68 g/kg GLSO and 50 mg/kg CTX; GLSO-M + CTX: mice were given 2.496 g/kg GLSO and 50 mg/kg CTX; GLSO-L + CTX: mice were given 0.312 g/kg GLSO and 50 mg/kg CTX. GLSO: Ganoderma Lucidum Spore Oil; CTX: cyclophos-phamide. Kaplan-Meier survival analysis was employed to compare the difference of survival rates between groups (n = 22-25).
Table 2 Thymus, spleen and liver indices in each group (mg/g, $\bar{x}±s$)
| Group | n | Thymus index | Spleen index | Liver index |
|---|---|---|---|---|
| Control | 6 | 0.226±0.045 | 4.711±0.638 | 42.531±10.011 |
| GLSO | 8 | 0.248±0.074 | 4.656±0.387 | 49.983±6.393 |
| CTX | 8 | 0.160±0.030a | 5.484±1.766 | 55.010±4.888a |
| Vehicle+CTX | 8 | 0.167±0.018a | 5.096±1.048 | 53.687±5.359d |
| GLSO-H+CTX | 8 | 0.232±0.040b | 5.292±0.784 | 52.048±6.397 |
| GLSO-M+CTX | 8 | 0.204±0.026b | 5.511±0.960 | 53.738±4.913 |
| GLSO-L+CTX | 8 | 0.246±0.046c | 4.763±0.399 | 51.727±6.517 |
Table 2 Thymus, spleen and liver indices in each group (mg/g, $\bar{x}±s$)
| Group | n | Thymus index | Spleen index | Liver index |
|---|---|---|---|---|
| Control | 6 | 0.226±0.045 | 4.711±0.638 | 42.531±10.011 |
| GLSO | 8 | 0.248±0.074 | 4.656±0.387 | 49.983±6.393 |
| CTX | 8 | 0.160±0.030a | 5.484±1.766 | 55.010±4.888a |
| Vehicle+CTX | 8 | 0.167±0.018a | 5.096±1.048 | 53.687±5.359d |
| GLSO-H+CTX | 8 | 0.232±0.040b | 5.292±0.784 | 52.048±6.397 |
| GLSO-M+CTX | 8 | 0.204±0.026b | 5.511±0.960 | 53.738±4.913 |
| GLSO-L+CTX | 8 | 0.246±0.046c | 4.763±0.399 | 51.727±6.517 |
Table 3 Plasma level of UREA, CRE, ALT, AST in each group ($\bar{x}±s$)
| Group | n | UREA (mmol/L) | CRE (μmol/L) | ALT (U/L) | AST (U/L) |
|---|---|---|---|---|---|
| Control | 7 | 6.8±1.0 | 16.0±1.4 | 194.9±18.7 | 1514.1±171.1 |
| GLSO | 7 | 5.3±1.3a | 14.0±2.2 | 195.0±79.1 | 1462.4±452.5 |
| CTX | 7 | 6.8±1.2 | 15.3±2.3 | 74.7±10.2b | 383.1±70.9b |
| Vehicle+CTX | 7 | 6.3±0.8 | 15.6±1.7 | 72.1±9.1b | 348.7±73.5b |
| GLSO-H+CTX | 7 | 5.4±0.5c | 15.3±2.8 | 73.9±12.7 | 383.0±58.1 |
| GLSO-M+CTX | 7 | 5.2±0.8c | 14.4±1.7 | 72.4±4.3 | 408.4±103.5 |
| GLSO-L+CTX | 7 | 5.8±1.4 | 14.1±2.3 | 69.0±12.7 | 366.9±60.1 |
Table 3 Plasma level of UREA, CRE, ALT, AST in each group ($\bar{x}±s$)
| Group | n | UREA (mmol/L) | CRE (μmol/L) | ALT (U/L) | AST (U/L) |
|---|---|---|---|---|---|
| Control | 7 | 6.8±1.0 | 16.0±1.4 | 194.9±18.7 | 1514.1±171.1 |
| GLSO | 7 | 5.3±1.3a | 14.0±2.2 | 195.0±79.1 | 1462.4±452.5 |
| CTX | 7 | 6.8±1.2 | 15.3±2.3 | 74.7±10.2b | 383.1±70.9b |
| Vehicle+CTX | 7 | 6.3±0.8 | 15.6±1.7 | 72.1±9.1b | 348.7±73.5b |
| GLSO-H+CTX | 7 | 5.4±0.5c | 15.3±2.8 | 73.9±12.7 | 383.0±58.1 |
| GLSO-M+CTX | 7 | 5.2±0.8c | 14.4±1.7 | 72.4±4.3 | 408.4±103.5 |
| GLSO-L+CTX | 7 | 5.8±1.4 | 14.1±2.3 | 69.0±12.7 | 366.9±60.1 |
Figure 3 Effect of GLSO and CTX on the protein expression level of PD-1, JAK2, p-STAT3 and STAT3 in spleen and PD-L1, JAK2, p-STAT3 and STAT3 in tumor A: PD-1, JAK2, p-STAT3, STAT3, and pSTAT3/STAT3 level in spleen; A1: PD-1 expression was detected by Western blot analysis; A2: JAK2 expression was detected by Western blot analysis; A3: p-STAT3 expression was detected by Western blot analysis; A4: STAT3 expression was detected by Western blot analysis; A5: Western blot analysis of PD-1 expression; A6: Western blot analysis of JAK2 expression; A7: Western blot analysis of p-STAT3 expression; A8: Western blot analysis of STAT3 expression; A9: Western blot analysis of p-STAT3/STAT3 level; B: PD-L1, JAK2, p-STAT3, STAT3, and pSTAT3/STAT3 level in tumor; B1: PD-L1 expression was detected by Western blot analysis; B2: JAK2 expression was detected by Western blot analysis; B3: p-STAT3 expression was detected by Western blot analysis; B4: STAT3 expression was detected by Western blot analysis; B5: Western blot analysis of PD-L1 expression; B6: Western blot analysis of JAK2 expression; B7: Western blot analysis of p-STAT3 expression; B8: Western blot analysis of STAT3 expression; B9: Western blot analysis of p-STAT3/STAT3 level. Control group: mice were given normal saline; GLSO group: mice were given 4.68 g/kg GLSO and normal saline; CTX group: mice were given normal saline and 50 mg/kg CTX; Vehicle + CTX: mice were given vehicle and 50 mg/kg CTX; GLSO-H + CTX: mice were given 4.68 g/kg GLSO and 50 mg/kg CTX; GLSO-M + CTX: mice were given 2.496 g/kg GLSO and 50 mg/kg CTX; GLSO-L + CTX: mice were given 0.312 g/kg GLSO and 50 mg/kg CTX. GLSO: Ganoderma Lucidum Spore Oil; CTX: cyclophosphamide; PD-1: programmed death-1; PD-L1: programmed death ligand 1; JAK2: janus kinase 2; p-STAT3: phosphorylated signal transducer and activator of transcription 3; STAT3: signal transducer and activator of transcription 3. Data are presented as the mean ± standard deviation using one-way analysis of variance (n = 5). aP < 0.05, bP < 0.01, dP < 0.001, vs Control group, cP < 0.05, eP < 0.001, vs Vehicle + CTX group.
Figure 3 Effect of GLSO and CTX on the protein expression level of PD-1, JAK2, p-STAT3 and STAT3 in spleen and PD-L1, JAK2, p-STAT3 and STAT3 in tumor A: PD-1, JAK2, p-STAT3, STAT3, and pSTAT3/STAT3 level in spleen; A1: PD-1 expression was detected by Western blot analysis; A2: JAK2 expression was detected by Western blot analysis; A3: p-STAT3 expression was detected by Western blot analysis; A4: STAT3 expression was detected by Western blot analysis; A5: Western blot analysis of PD-1 expression; A6: Western blot analysis of JAK2 expression; A7: Western blot analysis of p-STAT3 expression; A8: Western blot analysis of STAT3 expression; A9: Western blot analysis of p-STAT3/STAT3 level; B: PD-L1, JAK2, p-STAT3, STAT3, and pSTAT3/STAT3 level in tumor; B1: PD-L1 expression was detected by Western blot analysis; B2: JAK2 expression was detected by Western blot analysis; B3: p-STAT3 expression was detected by Western blot analysis; B4: STAT3 expression was detected by Western blot analysis; B5: Western blot analysis of PD-L1 expression; B6: Western blot analysis of JAK2 expression; B7: Western blot analysis of p-STAT3 expression; B8: Western blot analysis of STAT3 expression; B9: Western blot analysis of p-STAT3/STAT3 level. Control group: mice were given normal saline; GLSO group: mice were given 4.68 g/kg GLSO and normal saline; CTX group: mice were given normal saline and 50 mg/kg CTX; Vehicle + CTX: mice were given vehicle and 50 mg/kg CTX; GLSO-H + CTX: mice were given 4.68 g/kg GLSO and 50 mg/kg CTX; GLSO-M + CTX: mice were given 2.496 g/kg GLSO and 50 mg/kg CTX; GLSO-L + CTX: mice were given 0.312 g/kg GLSO and 50 mg/kg CTX. GLSO: Ganoderma Lucidum Spore Oil; CTX: cyclophosphamide; PD-1: programmed death-1; PD-L1: programmed death ligand 1; JAK2: janus kinase 2; p-STAT3: phosphorylated signal transducer and activator of transcription 3; STAT3: signal transducer and activator of transcription 3. Data are presented as the mean ± standard deviation using one-way analysis of variance (n = 5). aP < 0.05, bP < 0.01, dP < 0.001, vs Control group, cP < 0.05, eP < 0.001, vs Vehicle + CTX group.
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