Acteoside promotes autophagy and apoptosis of hepatoma cells by regulating JNK signaling pathway / 中国中药杂志

This study explored the molecular mechanism of acteoside against hepatoma 22(H22) tumor in mice through c-Jun N-terminal kinase(JNK) signaling pathway. H22 cells were subcutaneously inoculated in 50 male BALB/c mice, and then the model mice were classified into model group, low-dose, medium-dose, and high-dose acteoside groups, and cisplatin group. The administration lasted 2 weeks for each group(5 consecutive days/week). The general conditions of mice in each group, such as mental status, diet intake, water intake, activity, and fur were observed. The body weight, tumor volume, tumor weight, and tumor-inhibiting rate were compared before and after administration. Morphological changes of liver cancer tissues were observed based on hematoxylin and eosin(HE) staining, and the expression of phosphorylated(p)-JNK, JNK, B-cell lymphoma-2(Bcl-2), Beclin-1, and light chain 3(LC3) in each tissue was detected by immunohistochemistry and Western blot. qRT-PCR was performed to detect the mRNA expression of JNK, Bcl-2, Beclin-1, and LC3. The general conditions of mice in model and low-dose acteoside groups were poor, while the general conditions of mice in the remaining three groups were improved. The body weight of mice in medium-dose acteoside group, high-dose acteoside group, and cisplatin group was smaller than that in model group(P<0.01). The tumor volume in model group was insignificantly different from that in low-dose acteoside group, and the volume in cisplatin group showed no significant difference from that in high-dose acteoside group. Tumor volume and weight in medium-dose and high-dose acteoside groups and cisplatin group were lower than those in the model group(P<0.001). The tumor-inhibiting rates were 10.72%, 40.32%, 53.79%, and 56.44% in the low-dose, medium-dose, and high-dose acteoside groups and cisplatin group, respectively. HE staining showed gradual decrease in the count of hepatoma cells and increasing sign of cell necrosis in the acteoside and cisplatin groups, and the necrosis was particularly obvious in the high-dose acteoside group and cisplatin group. Immunohistochemical results suggested that the expression of Beclin-1, LC3, p-JNK, and JNK was up-regulated in acteoside and cisplatin groups(P<0.05). The results of immunohistochemistry, Western blot, and qRT-PCR indicated that the expression of Bcl-2 was down-regulated in the medium-dose and high-dose acteoside groups and cisplatin group(P<0.01). Western blot showed that the expression of Beclin-1, LC3, and p-JNK was up-regulated in acteoside and cisplatin groups(P<0.01), and there was no difference in the expression of JNK among groups. qRT-PCR results showed that the levels of Beclin-1 and LC3 mRNA were up-regulated in the acteoside and cisplatin groups(P<0.05), and the level of JNK mRNA was up-regulated in medium-dose and high-dose acteoside groups and cisplatin group(P<0.001). Acteoside promotes apoptosis and autophagy of H22 cells in mice hepatoma cells by up-regulating the JNK signaling pathway, thus inhibiting tumor growth.

He-Wei Granule enhances anti-tumor activity of cyclophosphamide by changing tumor microenvironment / 中草药·英文版

Objective: He-Wei Granule (HWKL) is a modern product derived from the modified formulation of traditional Chinese medicine Banxia Xiexin Decoction (BXD), which remarkedly enhanced the anti-proliferation activity of cyclophosphamide (CTX) on HepG2 and SGC-7901 cell lines in vitro in our previous research. The aim of the study was to investigate the synergistic effects of HWKL and CTX using a transplanted H22 hepatocellular carcinoma mouse model. Methods: The CTX-toxic-reducing efficacy of HWKL was evaluated by hematology indexes, organ indexes and marrow DNA detection. To investigate the underlying mechanisms, histopathology test, immunohistochemistry test and TUNEL staining were conducted. The efficacy of HWKL on the micro-vessel density (MVD) in tumor tissue was also evaluated by measuring CD34 level. Results: High dose HWKL (6.75 g/kg) markedly attenuated CTX-induced hepatotoxicity and myelosuppression while significantly enhanced CTX anticancer efficacy in vivo. Further mechanism investigation suggested that high dose HWKL significantly increased cleaved Caspase 3 level and promoted apoptosis in tumor tissue by up-regulating Bax expression and down-regulating Bcl-2 and FasL expressions. Compared with CTX alone group, the decrease in LC-3B and Beclin 1 levels suggested that the autophagy in H22 carcinoma was significantly inhibited with addition of high dose HWKL. ELISA assay results indicated that the autophagy inhibition was achieved by decreasing p53 expression, blocking PI3K/AKT/mTOR pathway and recovering Th1/Th2 cytokine balance. In addition, CD34 and EGFR immunohistochemistry assay suggest that high dose HWKL could significantly decrease micro-vessel density (MVD) and inhibit angiogenesis in H22 carcinoma. Conclusion: It can be concluded that high-dose HWKL enhanced CTX efficacy by promoting apoptosis, inhibiting autophagy and angiogenesis in tumor tissue while significantly alleviated CTX-induced toxicity, and could be applied along with CTX in clinical treatment as a supplement agent.

Effects of increasing efficacy and decreasing toxicity of ginkgo flavone aglycone on doxorubicin in the treatment of liver cancer / 中国药房

OBJECTIVE To study the effects of increasing efficacy and decreasing toxicity of ginkgo flavone aglycone （GA） on doxorubicin （DOX）in the treatment of liver cancer. METHODS A tumor bearing model was established by inoculating liver cancer cell H 22 into the right axillary skin of ICR mice. The successfully modeled mice were randomly divided into model control group，DOX group （2.5 mg/kg，once every other day ，via tail vein ），GA group （30 mg/kg，once a day ，gavage）and GA+DOX group（the usage was the same as single drug groups ），with 6 mice in each group. The administration cycle was 15 days. The general growth of mice in each group were observed ，body weight and tumor weight were measured ，and the inhibition rate of tumor was calculated. Jin’s formula was used to evaluate the effect of combined medication （Q）. The serum level of alpha-fetal protein（AFP），the pathological changes of tumor tissue ，cell apoptosis and the expression of platelet-endothelial cell adhesion molecule-1（CD31）were detected in each group. The cardiac index，serum levels of B-type natriuretic peptide （BNP）and N-terminal pro-brain natriuretic peptide （NT-pro BNP ），pathological changes of heart and myocardial fibrosis degree were also detected. RESULTS The percentage of body weight change （except for GA group ） and tumor weights of DOX group，GA group and GA + DOX group were all decreased significantly，compared with model control group （P＜0.05 or P＜0.01），while tumor weight of GA+DOX gro up was significantly lower than DOX group （P＜0.01）. Inhibitory rates of tumor in 3 administration groups were 54.29％，42.50％ and 89.29％ respectively，and Q of two-drug combination was 1.21. The tumor tissues of mice in each administration group were necrotic to varying degrees ；the serum level of AFP and the expression of CD31 in tumor tissue were decreased significantly ，compared with model control group （P＜0.05 or P＜0.01）；the percentage of necrosis area of tumor tissue and the positive rate of apoptosis （except for single drug groups ）were significantly increased （P＜0.05 or P＜0.01），while positive rate of apoptosis in GA+DOX group was significantly higher than DOX group （P＜0.05）. Cardiac index of mice in DOX group was significantly lower than model control group （P＜0.01）；serum levels of BNP and NT-pro BNP in DOX group and GA+ DOX group were significantly higher than model control group （P＜0.05 or P＜0.01）；pathological changes of heart and the degree of myocardial fibrosis in GA+DOX group were lower than DOX group. CONCLUSIONS GA combined with DOX show synergistic antitumor effect. GA can strengthen the apoptosis promoting effect of DOX ，and can help to reduce the cardiotoxicity of DOX.

Therapeutic effect of Panax notoginseng saponins combined with cyclophosphamide in mice bearing hepatocellular carcinoma H22 cell xenograft / 南方医科大学学报

OBJECTIVE@#To investigate the therapeutic effects of total saponins from Panax notognseng (PNS) combined with cyclophosphamide (CTX) in mice bearing hepatocellular carcinoma H22 cell xenograft.@*METHODS@#We examined the effects of treatment with different concentrations of PNS on H22 cell proliferation for 24 to 72 h in vitro using CCK8 colorimetric assay. Annexin V/PI double fluorescence staining was used to detect the effect of PNS on apoptosis of H22 cells. Mouse models bearing H22 cell xenograft were established and treated with CTX (25 mg/kg), PNS (120, 240 or 480 mg/kg), alone or in combinations. After treatments for consecutive 10 days, the mice were euthanized for examinations of carbon clearance ability of the monocytes and macrophages, splenic lymphocyte proliferation, tumor necrosis factor (TNF-α), interleukin-2 (IL-2), serum hemolysin antibody level, blood indicators, and the tumor inhibition rate.@*RESULTS@#Treatment with PNS concentration-dependently inhibited the proliferation and significantly promoted apoptosis of cultured H22 cells (P < 0.01). In the tumor-bearing mouse models, PNS alone and its combination with CTX both resulted in obvious enhancement of phagocytosis of the monocyte-macrophages, stimulated the proliferation of splenic lymphocytes, promoted the release of TNF-α and IL-2 and the production of serum hemolysin antibody, and increased the number of white blood cells, red blood cells and lymphocytes in the peripheral blood. Treatment with 480 mg/kg PNS combined with CTX resulted in a tumor inhibition rate of 83.28% (P < 0.01) and a life prolonging rate of 131.25% in the mouse models (P < 0.05).@*CONCLUSION@#PNS alone or in combination with CTX can improve the immunity and tumor inhibition rate and prolong the survival time of H22 tumor-bearing mice.

Sensitization of lidocaine on hepatoma H22-bearing mice treated by mitomycin / 肿瘤研究与临床

Objective To observe the sensitization of lidocaine on subcutaneous hepatoma H22-bearing mice and abdominal cavity H22 tumor-bearing mice treated by mitomycin. Methods According to the random number table method, the mice were divided into subcutaneous tumor-bearing group and abdominal cavity tumor-bearing group, with 15 mice in each group. The mice in the two groups were further divided into three subgroups: model group, mitomycin group, mitomycin+lidocaine group, with 5 mice in each subgroup. The day before the intraperitoneal injection, the density of H22 cells obtained from peritoneal culture of one mouse was adjusted to 5 ×106/ml. Subcutaneous tumor-bearing group mice were injected H22 cells into the right armpit, and abdominal cavity tumor-bearing group mice were injected H22 cells into the abdominal cavity, 0.2 ml per mouse. Intraperitoneal injection was given after inoculation for 24 h (the experiment day 1), followed by intraperitoneal injection on day 5 and 9. Univariate ANOVA analysis and t test were used to analyze the solid tumor weight and tumor inhibition rate on the 11th day of subcutaneous tumor-bearing mice, and the survival time and life extension rate within 60 days of abdominal cavity tumor-bearing mice. Results The solid tumor weight of subcutaneous tumor-bearing mice model group, mitomycin group and mitomycin + lidocaine group were (3.77 ±1.02) g, (1.67 ±0.28) g, (0.74 ±0.19) g, respectively, and the differences in the three groups were statistically different (F = 31.753, P < 0.01); compared with the subcutaneous model group, the subcutaneous solid tumor weights of mitomycin group and mitomycin +lidocaine group were decreased and the differences were both statistically different (t=2.10, P<0.01; t=3.04, P<0.01); the subcutaneous solid tumor weight of mitomycin+lidocaine group was lower than that of mitomycin group (t= 0.93, P= 0.034). The tumor inhibition rate of mitomycin group and mitomycin +lidocaine group reached 55.70％ and 80.37％ respectively. The survival time of abdominal cavity tumor-bearing mice in model group, mitomycin group and mitomycin + lidocaine group was (16.80±0.84) d, (28.80± 6.30) d, (40.40±12.86) d, respectively, and the differences in the three groups were statistically different (F=10.155, P=0.003); compared with the abdominal cavity tumor-bearing mice model group, the survival time of mice in mitomycin group and mitomycin + lidocaine group was prolonged (t= 12.00, P= 0.041; t= 23.60, P= 0.001), and it was found that survival time in mitomycin + lidocaine group was longer than that in mitomycin group (t=11.60, P=0.047). The life extension rate of mitomycin group and mitomycin+lidocaine group reached 71.43％ and 140.48％ respectively. Conclusion Lidocaine can increase the sensitization of mitomycin on hepatoma H22-bearing mice.

Effect of Modified Lichongtang Combined with 5-Fluorouracil on Epithelial Mesenchymal Transition in H22 Tumor-bearing Mice / 中国实验方剂学杂志

Objective: To investigate the effect of modified Lichongtang combined with 5-fluorouracil (5-FU) on tumor epithelial-mesenchymal transition (EMT) in H22 tumor-bearing mice. Method: Mouse models of transplanted hepatoma were constructed. After tumor formation, they were randomly divided into 4 groups:blank group, 5-FU group(2.5 mg·kg-1 5-FU intraperitoneal injection), modified Lichongtang combined with 5-FU group (5-FU+Chinese medicine group), and modified Lichongtang group (Chinese medicine group,25 g·kg-1 gavage),n=10 in each group. The effect of modified Lichongtang combined with 5-FU on the tumor inhibition rate of subcutaneous transplanted tumor was observed. The gene expression levels of E-cadherin,N-cadherin,Snail,and Twist in transplanted tumor were observed by Real-time PCR. The protein expression levels of E-cadherin,N-cadherin,Snail,and Vimentin were detected by using Western blot. Result: The tumor inhibiting rate was 59.18%,84.42%,and 10.39% respectively in 5-FU group, 5-FU+Chinese medicine group,and Chinese medicine group. All of these can inhibit the growth of liver cancer transplantation tumor, and the tumor inhibiting rate of 5-FU+Chinese medicine group was significantly higher than that in 5-FU group (PPPPPPPPPPPConclusion: Modified Lichongtang, 5-FU and their combination have inhibitory effect on the growth of transplanted tumors of hepatocarcinoma mice, and the combination of the two drugs can enhance the effect of chemotherapy and to some extent inhibit the toxicity of 5-FU. The mechanism may be related to the inhibition of liver cancer EMT.

Study on Anti-tumor Effects of Periplaneta americana Polypeptide PAP- 2 on H 22 Tumor-bearing Mice / 中国药房

OBJECTIVE： To investigate anti-tumor effects of Periplaneta americana polypeptide PAP-2 on H22 tumor-bearing mice. METHODS： The mice tumor-bearing model was established by subcutaneous injection of ascites of H22 hepatocellular carcinoma mice via axilla. 70 mice were randomly divided into model group （normal saline）， 5-FU group （positive drug control， 20 mg/kg）， P. americana extract skimmed cream group （200 mg/kg， calculated by extract）， CⅡ-3 group （polypeptide isolated from skimmed cream as main active ingredient， 200 mg/kg， calculated by extract） and polypeptide PAP-2 high-dose， medium-dose and low-dose groups （isolated from CⅡ-3， 200， 100， 50 mg/kg， calculated by monomer）， with 10 mice in each group. The mice in the 5-FU group were given intraperitoneal injection once every other day， while the mice in the other groups were given intragastric administration once a day， the administration cycle was 10 d. After medication， the changes of tumor were observed and the organs （spleen， thymus and liver） index were measured. Histopathological changes of tumor tissue were observed after HE staining. The contents of VEGF， IL-1β and IL-4 in serum were determined by ELISA. RESULTS： Skimmed cream， CⅡ-3 and different doses of PAP-2 could inhibit the growth of tumor in tumor-bearing mice to different extent and increase organ index， and PAP-2 showed a dose-effect relationship. The tumor inhibition rate （38.95％） of PAP-2 high dose group was significantly higher than those of skimmed cream group and CⅡ-3 group （P＜0.05）， which was close to that （40.87％） of 5-FU group （P＞0.05）. Spleen index， thymus index and liver index of mice in PAP-2 high dose group were significantly those of model group and CⅡ-3 group （P＜0.05）； and the liver index of mice in PAP-2 high dose group was significantly higher than that of skimmed cream group （P＜0.05）. In addition， PAP-2 could decrease the serum contents of VEGF and IL-4， and increased serum content of IL-1β， with high dose group showed significant difference compared with model group （P＜0.05）； the serum content of IL-1β of mice in PAP-2 high dose group was significantly higher the that of skimmed cream group and CⅡ-3 group （P＜0.05）， serum contnet of IL-4 in PAP-2 high dose group was significantly lower the  that of skimmed cream group and CⅡ-3 group （P＜0.05）， but the serum content in which was significantly lower than that of skimmed cream group and CⅡ-3 group（P＜0.05）. CONCLU- SIONS： P. americana polypeptide PAP-2 it has a certern anti-tumor effects on H22 tumor-bearing mice， and its can increase the index of organs of H22 tumor-bearing mice， decrease the contents of VEGF and IL-4 in serum， increase the content of IL-1β in serum.

Effect on tumor growth inhibition activity of Gekkoswinhonis Guenther with different extent of broiling on H22 tumor-bearing mice / 药学学报

This study was designed to investigate the effect on tumor growth inhibition activity of lizards (Gekkoswinhonis Guenther) with different extent of broiling. Samples were prepared by a traditional drying method combined with broiling on clay tiles. Four groups of samples were all dried before broiling. Group A was without broiling; group B was mildly broiled; group C was moderately broiled; and group D was heavily broiled. Crispiness was detected by the sizes of the generated fragments of different groups and crispiness increased with broiling. Sensory evaluation of vision and olfaction was performed, and scores were generated by evaluators. Moderately broiled group had the highest total score in sensory evaluation. Water content and content of water-soluble extracts were detected according to Chinese Pharmacopoeia. With the increasing broiling extent, content of water-soluble extracts increased while water content decreased. Soluble protein concentration was detected by bicinchoninic acid (BCA) kit and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with the same crude drug content. Soluble protein concentration decreased with the increasing broiling extent. With equal loading of proteins at the same concentration, soluble protein diversity was detected by SDS-PAGE. Band difference was marked by red boxes. Soluble protein molecule weights showed significant difference with the increasing broiling extent. H22 tumor-bearing mice model was established and used to detect tumor growth inhibition rate and immune organ index. Life quality of mice was evaluated. Mice treated with Gekkoswinhonis Guenther had better appetites and higher average weights compared with positive control group treated with fluorouracil (5-FU). Animal experiments were approved by the Ethics Committee of Beijing University of Chinese Medicine. Group A had the highest tumor growth inhibition rate (34.11%), followed by Group B (29.14%) and Group D (28.43%), Group C (21.98%) had the lowest tumor growth inhibition rate, but sensory evaluation was on the contrary. These results indicated that moderately broiling improved sensory evaluation but reduced the tumor growth inhibition activity of Gekkoswinhonis Guenther. The best tumor growth inhibition activity appeared when water content was 7.71%.

Anti-tumor and immune-modulating effect of decoction in mice bearing hepatoma H22 tumor / 南方医科大学学报

OBJECTIVE@#To investigate the antitumor activity of decoction and study its liver and kidney toxicity and its effect on the immune system in a tumor-bearing mouse model.@*METHODS@#Hepatoma H22 tumor-bearing mouse models were randomized into model group, cyclophosphamide (CTX) group, and low-, moderate-, and high-dose decoction groups (JW-L, JW-M, and JW-H groups, respectively). The antitumor activity of decoction was assessed by calculating the tumor inhibition rate and pathological observation of the tumor tissues. Immunohistochemistry was used to detect the expressions of Bax, Bcl-2, Bax/Bcl-2 and caspase-3 in the tumors. The liver and kidney toxicity of decoction was analyzed by evaluating the biochemical indicators of liver and kidney functions. The immune function of the tumor-bearing mice were assessed by calculating the immune organ index, testing peripheral blood routines, and detection of serum IL-2 and TNF-α levels using enzyme-linked immunosorbent assay.@*RESULTS@#Compared with that in the model group, the tumor mass in CTX, JW-M and JW-H groups were all significantly reduced ( < 0.05) with cell rupture and necrosis in the tumors. Immunohistochemistry revealed obviously up-regulated expressions of Bax and caspase-3 and down- regulated expression of Bcl-2 protein with an increased Bax/Bcl-2 ratio in CTX, JW-M and JW-H groups. Treatment with decoction significantly reduced Cr, BUN, AST and ALT levels, improved the immune organ index, increased peripheral blood leukocytes, erythrocytes and hemoglobin levels, and up-regulated the levels of TNF-α and IL-2 in the tumor-bearing mice. These changes were especially significant in JW-H group when compared with the parameters in the model group ( < 0.01).@*CONCLUSIONS@# decoction has a strong anti-tumor activity and can improve the liver and kidney functions of tumor-bearing mice. Its anti-tumor effect may be attributed to the up-regulation of Bax, caspase-3, TNF-α and IL-2 levels and the down-regulation of Bcl-2 expression as well as the enhancement of the non-specific immune function.

A Research on the Anti-tumor Effects of Acid-Soluble Polysaccharides from Gloeostereum incamatum in H22 Tumor Bearing Mice / 世界科学技术-中医药现代化

This study aimed at exploring the inhibitory effect behind its mechanism on acid-soluble polysaccharides from G.incamatum in transplanted H22 tumor mice.Different indices,including tumor inhibitory rate,organ index of liver,thymus and spleen,IL-2,IFN-γ and TNF-α were detected for the evaluation of anti-tumor effects and the mechanism.Furthermore,HE staining and TUNEL assay were adopted to investigate the pathological changes of tumor tissue and cell apoptosis,respectively.As a result,the three dose groups of acidsoluble polysaccharides of G.incamatum successfully inhibited the proliferation of tumor cells,while organ indexes of spleen and thymus were improved and serum IL-2,IFN-γ and TNF-α increased.H&E staining and TUNEL assay showed the polysaccharides induced cell apoptosis,playing a significant role in the inhibition of tumor growth.In conclusion,acid-soluble polysaccharides of G.incamatum possessed significant anti-tumor effects,behind which the mechanism could be related to the regulation of immune regulation,cell apoptosis,and the protection of liver function.

Effects and mechanism of anti IL-9 antibody on malignant ascites of hepatic car-cinoma / 中国免疫学杂志

Objective:To explore the effects and mechanism of anti IL-9 antibody on malignant ascites ( MA) of hepatic carci-noma in mice.Methods:A mouse model of MA was established by mouse H 22 cell line.45 mice were divided randomly into experi-mental group,negative control group and blank control group at 24 hours after modeling,with 15 mice in each group.The experimental group was injected intraperitoneally with anti IL-9 antibody;the negative control group was injected with isotype IgG antibody;the blank control group was injected with normal saline .The weight and behavior of the mice were measured before each injection .Five mice of each group was sacrificed at 24 hours after the last injection to measure the volume of MA .The level of VEGF,MMP-2,IL-9 and IFN-γin MA were determined with ELISA assay;the survival time of rest mice were recorded and compared .Results:The mean volume of MA of experimental group,negative control group and blank control group were (6.70±1.52)ml,(10.28±1.75)ml,(10.36±2.30) ml,respectively,the MA volume of experimental group were lower as compared to negative control group and blank control group ( P0.05 ) .Conclusion:Intraper-itoneal injection anti IL-9 antibody on H22 ascites-bearing mice can effectively inhibit the generation of the MA .The mechanism may be related to the inhibition of the expression of the VEGF and IL-9.

Anti-tumor Effect of Ginsenoside Rg1 Pyrolysis Products (HPPRg1) on H22 Tumor-bearing Mice / 中国药学杂志

OBJECTIVE: To investigate the antitumor effect and molecular mechanism of ginsenoside Rg1 pyrolysis products (HPPRg1) on H22 tumor bearing mice. METHODS: To establish tumor model of transplanting H22 tumor-bearing mice and observe the anti-tumor effects of HPPRg1, H22 tumor-bearing mice were randomly divided into groups of control, model, cyclophosphamide (CTX, 30 mg·kg-1), low dosage of HPPRg1 (HPPRg1-L, 10 mg·kg-1), middle dosage of HPPRg1 (HPPRg1-H, 20 mg·kg-1) and high dosage of HPPRg1 (HPPRg1-H, 40 mg·kg-1) groups, respectively. Through evaluating inhibition rates of tumors, organ indices, and levels of TNF-α, IFN-γ and IL-2 to observe the anti-tumor effect of HPPRg1. In addition, H&E and Hoechst 33258 straining were used to observe the apoptosis of H22 tumor cell. RESULTS: Compared with the model group, the three dose groups of HPPRg1 can inhibit tumor proliferation. Mainly through the inhibition of tumor cell proliferation and pro-apoptosis to exert anti-tumor effect. CONCLUSION: HPPRg1 has a significantly inhibitory effect on H22 tumor-bearing mice, the mechanism may related to promote apoptosis of tumor cells and improve immunity.

Effects of volatile components in Oroxyli Semen on H22 bearing mice and its possible mechanisms / 中国药理学通报

Aim To study the inhibitory effect of volatile components in Oroxyli Semen on liver cancer and its possible mechanisms.Methods H22 bearing mouse model was used,the mice were divided into six groups:blank,model,positive (cytoxan,100 mg · kg-1),low-,mid-,and high-dose (17.5,35,and 70 mg · kg-1) volatile components groups,and then the mice were ig given once daily for consecutive 12 d.Then the tumor growth inhibitory rate,spleen and thymus indexes were calculated;the serum levels of IL-2,IL-6 were determined.HE staining was used to study of the apoptosis of the solid tumor.After treatment of SMMC-7721 cells with 0 ～ 1 g · L-1 of volatile components for 24,48 and 72 h,MTT assay was used to examine the proliferation.TUNEL method was applied to detect cell apoptosis,and RT-PCR method to detect Bax,Bcl-2,caspase-3 mRNA experssion.Results The inhibitory rate of volatile components high-dose on H22 bearing mice was 42.08％.The thymus index and the contents of serum IL-2 and IL-6 of H22 bearing mice were significantly higher than those in model group.Volatile components could significantly inhibit proliferation and induce apoptosis of SMMC-7721 cells,downregulate the expression of Bcl-2 mRNA,and up-regulate the expression of Bax,caspase-3 mRNA.Conclusions The volatile components in Oroxyli Semen have obvious anti-tumor activity in vitro and in vivo,and its mechanism may be related to enhancing immune system and promoting tumor cell apoptosis.

Improved antitumor efficacy by combinationtreatment with recombined VEGF protein vaccineand cyclophosphamide in H22 hepatocellular carcinoma bearing-mice / 中国药理学通报

Aim To investigate the antitumor and antiangiogenic effects of combined low-dose cyclophosphamide(CTX)and recombined VEGF protein vaccine.Methods In this experiment,H22 hepatocellular carcinoma model was established in BALB/c mice.Mice were randomly divided into four groups: control group,CTX group(CTX),VEGF protein vaccine group(V2)and CTX plus V2 group(CTX+V2).The anti-tumor efficacy and antiangiogenic effect were investigated using a subcutaneous tumor model and an intradermal tumor model.Western blot and ELISAwere further adopted to detect the specific anti-VEGF antibody.Results CTX+V2 group displayed a lower tumor volume and tumor weight than either the single therapy group in the subcutaneous tumor model(P<005 vs V2,P<001 vs CTX).Meanwhile,CTX+V2 was more effective for antagonizing tumor-associated angiogenesis compared with either the single therapy(P<005 vs V2,P<001 vs CTX).After CTX+V2 immunization,high titer of anti-VEGF antibody was detected by ELISA and verified by Western blot.Conclusion The therapy of CTX combined with V2 has significant synergistic effect against H22 hepatocellular carcinoma.

Effects of Rhaponticum uniflorum on angiogenesis and apoptosis of H22 transplanted tumor tissue in mice / 中国药学杂志

OBJECTIVE: To investigate the effects of Rhaponticum uniflorum extract (RUE) on angiogenesis and apoptosis of H22 transplanted tumor tissue in mice. METHODS: Mice bearing H22 hepatoma cells were randomly assigned into 5 groups: model, high, medium and low dose RUE, and 5-fluorouracil (5-Fu) group. The intervention was lasted for 10 d. The pathological changes were detected with hematoxylin & eosin (HE) staining, the apoptosis of hepatoma cells were determined by DNA laddering method, the microvessel densities (MVD) were detected using immunohistochemical assay, and the protein expression of vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 2 (VEGFR2) and hypoxia-inducible factor-1α (HIF-1α) were detected by Western blot method. RESULTS: RUE treatment reduced the cell proliferation, aggravated the necrosis of transplanted tumor tissue, reduced DNA fragmentation of H22 hepatoma cells, decreased MVD of tumor tissue, and down-regulated the protein expression of VEGF, VEGFR2 and HIF-1α of the transplanted tumor tissue, as compared with the model group. CONCLUSION: RUE could exhibit anti-angiogenic and pro-apoptotic effects against H22 hepatoma cells in mice, and its anti-angiogenic mechanism is probably related to down-regulation of VEGF, VEGFR2 and HIF-1α proteins.

Antitumor effect of alcohol extract of pileostegia tomentella on H22 tumor-bearing mice / 中国药学杂志

OBJECTIVE: To study the antitumor effect of Pileostegia tomentella 95% alcohol extract (PTAE) on H22 tumor-bearing mice and its possible mechanisms. METHODS: Sixty mice were chosen and mouse models bearing H22 solid tumor were established in fifty mice, and the others were as normal control. H22 tumor-bearing mice were randomly divided into five groups:model control group, fluorouracil group(20 mg·kg-1), PTAE high, middle and low-dose group(180, 90, 45 g·kg-1 of crude drug, respectively). The mice in treatment groups were intragastric administration respectively, meanwhile, the mice in normal control and model groups were treated with the same volume of distilled water, once a day for ten days. The blood was collected from eyeball in all mice, and the serum were separated and detected by ELISA for IL-2 and TNF-α. Then the mice were put to death. Their tumors, thymuses and spleens were separated and weighted, and the tumor inhibitory rates, thymus and spleen indexes were calculated. The pathological change of tumor tissue was observed. RESULTS: Compared with model control group, the tumor weights of PTAE high and middle-dose groups were significantly decreased (P<0.05, P<0.01), the tumor inhibitory rates were 37.44% and 38.46% respectively. The spleen index of PTAE middle-dose group was increased significantly (P<0.01). The level of IL-2 in serum of tumor-bearing mice in the PTAE high-dose group was increased significantly(P<0.05), and the level of TNF-α in serum (P<0.01) could be increased significantly in the PATE high, middle and low-dose groups. CONCLUSION: Pileostegia tomentella 95% alcohol extract has antitumor activity, its mechanism may be developed by immuno-regulation.

Research on inhibition mechanism of polypeptide extract from scorpion venom combined with Rapamycin on angiogenesis of H22 hepatoma / 中草药

Objective: To study the inhibition of polypeptide extract from scorpion venom (PESV) combined with Rapamycin (RAPA) on angiogenesis of H22 hepatoma and its mechanism. Methods: The H22 carcinoma cell suspension was subcutaneously inoculated into 40 Kunming mice. Then tumor-bearing mice were randomly divided into four groups, i.e., control group, PESV group (20 mg/kg), RAPA group (2.5 mg/kg), and PESV + RAPA group, 10 mice in each group. The intervention was lasted 14 d. The tumor volume was measured once every other day, the tumor volume growth curve was drawn, and the tumor inhibitory rate was calculated. The protein expression levels of mammal target of RAPA (mTOR), hypoxia-inducible factor-1α (HIF-1α), and vascular endothelial growth factor-A (VEGF-A) were detected by immunohistochemical assay. Microvessel was signed by factor VIII and then detected by immunohistochemical assay. Results: The growth speed of H22 hepatoma transplantation tumor was obviously inhibited in the PESV group, RAPA group, and PESV + RAPA group compared with control group after 14 d intervention (P 22 hepatoma, the mechanism maybe related to the inhibition on the protein expression of mTOR, HIF-1α, and VEGF-A.

The anti-tumor effect of Coix stalk alcohol extraction on H22 tumor-bearing mice / 天津医药

Objective To study the anti-tumor effects of alcohol extraction of Coix stalk objects on H22 tumor-bearing mice. Methods The animal model of tumor bearing mice with H22 ascitic tumor cells was established. Eighty-four model mice were randomly and equally divided into Coix stalk extract groups 1-5 (10, 8, 6, 4 and 2 g/kg), model control group and cyclophosphamide group. Mice were treated orally with Coix stalk alcohol extraction solution (10, 8, 6, 4 and 2 g/kg), cyclophosphamide 0.02 g/kg and normal saline once a day for 8 days for Coix stalk extract group, cyclophosphamide group and model control group. The mouse activity, the size and the appearance of time of abdominal swelling, and changes of hair, feeding and drinking water quantity were observed in groups of mice. The solid tumor mass was measured in H22 tumor-bearing mice. The tumor inhibitory rate, liver index, spleen index and thymus index were calculated. Results The axillary tumor muster was found first in model control group with the fastest growth, reduced independent activity, decreased appetite and dim in hair color, followed by the Coix stalk extract group 1 and group 2. The last was Coix stalk extract group 5 and cyclophosphamide group. The solid tumor mass were (0.47±0.18), (0.37± 0.13), (0.34±0.10), (0.30±0.11) and (0.28±0.09) mg for Coix stalk alcohol extract groups 1-5, which were significantly lower than those of model control group (0.60 mg±0.21 mg, F=5.700,P<0.05). The tumor inhibition rates were 21.67%, 38.33%, 43.33%, 50.00%, 53.33%and 60.00%in Coix stalk extract groups 1-5 and cyclophosphamide group. The liver index, spleen index and thymus index were lower in cyclophosphamide group and Coix stalk alcohol extract groups than those of model control group (except for the spleen index of Coix stalk extract group 1). The liver index was lower in Coix stalk ethanol extract groups than that of cyclophosphamide group. There were no significant differences in the spleen index, thymus index between Coix stalk ethanol extract groups and cyclophosphamide group. Conclusion Coix stalk alcohol extract has inhibitory effects on the tumor and liver damage in H22 mice.

Pharmacodynamics Study of Different Lactone Ratios of Hydroxycamptothecin on Mice Models of H22 Hepatoma / 医药导报

Objective To investigate the anti-tumor effects of hydroxycamptothecin ( HCPT ) with different lactone ratios on the mice models of H22 hepatoma. Methods Mice models of H22 hepatoma were established. Tumor inhibiting rates of HCPT with different lactone ratios ( 0%, 25%, 50%, 75%, 100%) and the growth status of model mice before and after chemotherapy were observed. Serum biochemical indices were determined to investigate the effects of HCPT with different lactone ratios on hepatic and renal function of the mice. Results Positive control drug and HCPT with different lactone ratios all inhibited the tumor in mice with H22 hepatoma, the inhibition rate was 65. 30%, 12. 57%, 49. 23%, 75. 47%, 90. 06% and 93. 22%, respectively. Compared with the model control group, the living conditions of the mice in HCPT groups were improved. With increasing of lactone ratios, the hepatic injury was alleviated markedly, but the renal injury was aggravated. Conclusion There is a positive correlation between lactone ratios and its anti-tumor effect, and HCPT with 75% lactone can achieve preferable anti-tumor effect with less toxicity as compared with that with 100% lactone ratio.

Preliminary study on the inhibitory effect of salidroside on H22 ascites hepatoma / 中国药理学通报

Aim To investigate the inhibitory effect of salidroside on tumor growth in tumor-bearing mice and explore its possible mechanisms of anti-tumor action in vivo. Methods The tumor-bearing mice model was established, and the mice were randomly divided into five experimantal groups, including the salidroside low, medium and high doses groups, NS group and positive control matrine group. After continuous injec-tion of drug for eight days, the tumor growth and sur-vival time were measured, tumor growth inhibition rate and life extension rate, spleen index,the content of se-rum IL-2 and IL-12 were calculated. Results The general situation of the salidroside mice was better than the NS group and the positive control matrine group. Compared with NS group, the salidroside low,medium and high doses groups showed inhibitory effects on H22 solid tumor ( P<0. 01 ) with inhibition ratio 28. 9 %, 43. 3 % and 53. 5 %,and lengthened the survival time of mice ( P<0. 01 ) with the life extension rate 37. 54%,48. 96 % and 52. 85 % respectively. The spleen index and the content of serum IL-2 and IL-12 of the salidroside mice were significantly higher than NS group and matrine group. Conclusion Salidroside has marked inhibitory effects on tumor growth in vivo, its mechanism is probably related to improving the anti-tumor immune function.