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.

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.

Study on antidepressant effect of total alkaloids of Fibraurea recisa / 中国中药杂志

This study aimed to investigate the antidepressant effects of total alkaloids of Fibraurea recisa in HT22 cells damaged by corticosterone (CORT) in vitro and in a mouse model of chronic unpredictable mild stress (CUMS) as well as the underlying mechanisms.In cellular experiments,the viability of CORT-damaged HT22 cells was detected using cell counting kit-8 (CCK-8),and the cell apoptosis was detected by Hoechst 33258 staining.In animal experiments,C57BL/6N mice were randomly divided into the control group,model group,low (100 mg·kg~(-1)),medium (200 mg·kg~(-1)) and high (400 mg·kg~(-1))-dose of total alkaloids of F.recisa groups,and positive control group.After 21 days of CUMS exposure,their depressive behaviors were observed in behavioral and Morris water maze tests.The serum levels of 5-hydroxytryptamine (5-HT),dopamine (DA),and norepinephrine (NE) were assessed by ELISA.The expression levels of apoptosis-related proteins Bcl-2,Bax and cleaved caspase-3 in HT22 cells and mouse hippocampus were detected by Western blot.The results suggested that total alkaloids of F.recisa alleviated the damage of HT22 cells induced by CORT in a dose-dependent manner.The Hoechst 33258 staining uncovered that total alkaloids of F.recisa better reduced the blue spots and inhibited cell apoptosis.The results of animal experiments showed that total alkaloids of F.recisa significantly improved the depression-like behaviors of mice and increased the serum levels of 5-HT,DA and NE as compared with those in the model group.The Western blot assays revealed a significant up-regulation of Bcl-2 protein expression,but an obvious reduction in Bax and cleaved caspase-3protein expression in the total alkaloids of F.recisa group.In conclusion,total alkaloids of F.recisa inhibited depression possibly by regulating the apoptosis-related protein expression or elevating the monoamine neurotransmitter levels in the brain.

Study on Protective Effects and Mechanism of Lespedeza cuneata Extracts on Glutamate-induced Hippocampal Cells HT 22 Injury of Mice Based on Nrf 2/HO-1 Signaling Pathway / 中国药房

OBJECTIVE：To study the protective effects of Lespedeza cunea ta extract on glutamate-induced hippocampal cells HT22 injury of mice and its possible mechanism based on Nrf 2/HO-1 signaling pathway. METHODS ：Using glutamate （5 mmol/L） to extablish the injury model of HT 22 cells. Using water soluble vitamin E as positive control （50 µmol/L），MTT assay was used to detect the effects of 0（blank control ），25，50，100 µg/mL petroleum ether extract ，dichloromethane extract ，ethyl acetate extract of L. cuneata on the proliferation of glutamate-induced injury cellsafter pretreated for 12 h. Using water soluble vitamin E as positive control （50 µmol/L），DCFH-DA assay was used to detect the effects of 0（blank control ），25，50，100 µg/mL L. cuneata dichloromethane extract on the level of active oxygen （ROS）in glutamate-induced injury cells after pretreated with 12 h. Using HO-1 agonist CoPP as positive control ，Western blotting method was used to detect the effects of 0（blank control ），25，50，100 µg/mL L. cuneata dichloromethane extract on the protein expression of HO- 1 after treated for 24 h. Western blotting method （treated for 0.5，1，1.5 h）and immunofluorescence staining （treated for 1 h）were used to detect the effects of 100 µg/mL L. cuneata dichloromethane extract on protein expression of Nrf 2 inside and outside the nucleus. After HO-1 gene was silenced by small interfering RNA （Si RNA ）transfection technology ，the effects of 100 µg/mL L. cuneata dichloromethane extract on the survival rates of glutamate-induced injury cells and the level of ROS were detected. RESULTS ：Compared with blank control ，50， 100 µg/mL L. cuneata dichloromethane extract could significantly improve the survival rate of glutamate-induced injury cells （P＜ 0.05），while reduced the level of ROS （P＜0.05）. 25，50， 100 µg/mL L. cuneata dichloromethane extract could increase the protein expression of HO- 1 in cells（P＜0.05），while 100 com µg/mL L. cuneata dichloromethane extract could significantly decrease the protein le vel of Nrf 2 in cytoplasm and increasethat in nucleus （P＜0.05）. After HO-1 gene silencing ，the effects of L. cuneata dichloromethane extract on the proliferation promotion of glutamate-induced injury cells and the reduction of ROS level were reversed （P＜0.05）. CONCLUSIONS ：L. cuneata dichloromethane extract can protect HT 22 cells against injury induced by glutamate through activating Nrf 2 pathway，inducing HO- 1 expression.

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.

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%.

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.

Source analysis of lymphocytes secreting interleukin-22 from spleen of mice infected with Trichinella spiralis at the early encapsulated stage / 中华地方病学杂志

Objective To investigate the cell origin of interleukin (IL)-22-secreting cell of mice infected with Trichinella spiralis (T.spiralis) at the early encapsulated stage.Methods Twelve Balb/c mice were divided into the infected group and the control group according to body weight by random number table.The infected mice were intragastrically administrated with 300 muscle larvae of T.spiralis,and the control mice were given the same amount of normal saline.The IL-22-secreting cell subsets in mouse splenic lymphocytes were detected by flow cytometry at the fourth week after infection.Results The proportion of IL-22-secreting cells in splenic lymphocytes of T.spiralis infected mice was increased when compared with control group [(0.88 ± 0.25)％ vs (0.28 ±0.17)％,t =-4.899,P ＜ 0.05].There was no significant difference between the proportion of CD3+IL-22+ cells and CD3-IL-22+ cells in the splenic lymphocytes of the infected group [(0.29 ± 0.17)％ vs (0.51 ± 0.17)％,t =-2.195,P ＞ 0.05],and the percentage of CD3-IL-22+ cells were similar between the infected group and the control group [(0.51 ± 0.17)％ vs (0.44 ± 0.22)％,t =-0.600,P ＞ 0.05].The proportion of CD3+IL-22+ cells in the infected group was significantly higher than that in the control group [(0.29 ± 0.17)％ vs (0.07 ± 0.06)％,t =-3.068,P ＜ 0.05],and the percentage of CD4+IL-22+ T cells and γδTCR+IL-22+ T cells were obviously increased in CD3+ lymphocytes [(1.28 ± 0.54)％ vs (0.16 ± 0.07)％,(0.33 ± 0.22)％ vs (0.02 ± 0.00)％,t =-4.997,-3.342,P ＜ 0.05].Conclusions The proportion of IL-22-secreting splenic lymphocytes is increased in mice infected with T.spiralis at the early encapsulated stage.The rise is caused by increased numbers of IL-22-secreting CD3 + lymphocytes,especially CD4+ T cells and γδT cells.

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.

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.

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.

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.

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.

Effects of Sishen Pill and Gegen Qinlian Tablet on Inflammatory Cytokines in Mice with Colitis / 中国中医药信息杂志

Objective To explore the effects of Sishen Pill and Gegen Qinlian Tablet on cytokines of acute and chronic colitis induced by dextran sulfate sodium (DSS) in mice. Methods Mice freely drank 4%DSS dissolved in drinking water for continuous 5 days to establish acute colitis model. Mice circularly drank 3% DSS for 4 times for the establishment of chronic colitis model. In the acute or chronic colitis experiment, mice were randomly divided into control group, acute and chronic model groups, Sishen Pill group, and Gegen Qinlian Tablet group. Acute model group was administrated one day after DSS drinking for 8 days. Chronic model group was administrated after the second time of circular drinking for 16 days. ELISA was used to detect the contents of IFN-γ, IL-17 and IL-22 in cultural supernatant of mouse colon. Results Contents of IFN-γ, IL-17A and IL-22 in cultural supernatant increased significantly in acute models (P<0.01). Gegen Qinlian Tablet can significantly decrease the contents of IFN-γ and IL-17A of acute models (P<0.05). Sishen Pill can significantly decrease the content of IL-17A (P<0.05). IFN-γand IL-17A in cultural supernatant in chronic model mice significantly increased compared with control group (P<0.01). Sishen Pill and Gegen Qinlian Tablet inhibited the contents of IFN-γ and IL-17A. Compared with control group, Sishen Pill significantly increased the content of IL-22 (P<0.05). Conclusion Sishen Pill and Gegen Qinlian Tablet can treat colitis by decreasing the contents of IFN-γ and IL-17A in acute colitis model mice;Sishen Pill can treat chronic colitis by promoting IL-22 to increase.

Establishment and verification of a KCL22/NOD-SCID mouse transplantation tumor model of chronic myeloid leukemia / 中国实验动物学报

Objective To investigate the potential of chronic myeloid leukemia ( CML) cell line KCL22 in indu-cing leukemia in NOD-SCID mice for setting up a basis for constructing a CML mouse transplantation tumor model. Methods 2 ×107 KCL22 cells in logarithmic growth phase were injected via the tail vein into experimental NOD-SCID mice whereas PBS was injected to the mice of control group.General condition of the mice of both groups was observed.Wright staining was used to observe the changes of blood and bone marrow smears.PCR was conducted to detect the transcription level of BCR-ABL, and histology with HE staining was used to evaluate the tumor cell invasion in the liver and spleen. Results Four weeks after the injection of KCL22 cells, the mice in experimental group showed physical signs of decreased reactivity, depression, swollen hindlimb muscles and petechia on the hindlimb femur.Peripheral white blood cells ( WBC) began to increase after 5 weeks, with a significantly increased quantity compared with the control group (P90 days) (P<0.05).Conclusions A NOD-SCID mouse model of CML transplantation tumor is successfully established with leukemia KCL22 cells.

Evaluation of protective effect of IL-22 and IL-12 on cutaneous leishmaniasis in BALB/c mice

OBJECTIVE@#To investigate the protective effect of IL-22 and IL-12 on cutaneous leishmaniasisin BALB/c mice.@*METHODS@#The protective effect of IL-22 and IL-12 on cutaneous leishmanias in BALB/c mice was evaluated by measurement of IL-4, INF-γ, total IgG, IgG1 and IgG2a after challenge with Leishamania major. Clinical evaluations were performed by measurement of lesion diameter, and survival rate of the mice.@*RESULTS@#In week 27 post infection, the mortality rates for control groups were 100%. While the survival rates for the IL-12, IL-12 + IL-22, and IL-22(5 ng/g) groups were 100%. The size of lesions decreased in the presence IL-22 (5 ng/g) of mice weight, which was statistically significant in comparison with other groups (P<0.05). Mean of total IgG, IgG1 and IgG2a for IL-22 (5 ng/g) group was more than other groups. In IL-22 group (5 ng/g), INF-γ production was significantly higher than other groups and IL-4 was significantly lower than other groups.@*CONCLUSIONS@#The results obtained indicate the effectiveness of IL-22 and its effect on IL-12 in protection of cutaneous leishmaniasis.

Evaluation of protective effect of IL-22 and IL-12 on cutaneous leishmaniasis in BALB/c mice

Objective: To investigate the protective effect of IL-22 and IL-12 on cutaneous leishmaniasisin BALB/c mice. Methods: The protective effect of IL-22 and IL-12 on cutaneous leishmanias in BALB/c mice was evaluated by measurement of IL-4, INF-γ, total IgG, IgG1 and IgG2a after challenge with Leishamania major. Clinical evaluations were performed by measurement of lesion diameter, and survival rate of the mice. Results: In week 27 post infection, the mortality rates for control groups were 100%. While the survival rates for the IL-12, IL-12 + IL-22, and IL-22(5 ng/g) groups were 100%. The size of lesions decreased in the presence IL-22 (5 ng/g) of mice weight, which was statistically significant in comparison with other groups (. P<0.05). Mean of total IgG, IgG1 and IgG2a for IL-22 (5 ng/g) group was more than other groups. In IL-22 group (5 ng/g), INF-γ production was significantly higher than other groups and IL-4 was significantly lower than other groups. Conclusions: The results obtained indicate the effectiveness of IL-22 and its effect on IL-12 in protection of cutaneous leishmaniasis.

The study of xiaoaiping injection and octreotide acetate on anti-mouse H22 hepatocellular carcinoma and immune regulation / 中国医师杂志

Objective To observe the inhibitory effect of xiaoaiping injection and octreotide on H22 tumor-bearing mice and find the best drug concentration,then to explore its mechanism.Methods Establish a mouse H22 subcutaneous tumor model.After tumor the experiment animals were divided into normal control group,model group,Xiaoaiping low,medium and high dose group,octreotide group,and the group of XAP low,medium and high dose groups were combined with OCT.Calculate the tumor's volume and draw the tumor growth curve.Intraperitoneal injection for 14 days,Inhibitory rate was calculated; To observe its pathological changes by light microscope; The ratio.of CD3 + NK1.1-T cells,CD3-NK1.1 + NKcells,CD3 + NK1.1 + NK-Tcells in peripheral blood were measured by flow cytometry.Results Compared with the control group,H22 liver cancer in different treatment group had a certain inhibition effect on growth,The inhibitory effect of the combination group was better than single-agent group,High-dose Xiaoaiping + octreotide was best,Tumor model group compared with normal control group,The ratio of T cells,NK cells and NKT cells was significantly lower(P ＜0.05) ; T cells,NK cells and NKT cells after treatment in each group had some enhancement,High-dose Xiaoaiping + octreotide was the most obvious,the ratio of T cells,NK cells and NKT cells of the combination group was significantly more than the single-drug group and the same concentration of octreotide monotherapy Xiaoaiping group(P ＜ 0.05).Conclusion High-dose Xiaoaiping + octreotide is the best drug for the inhibitory drug concentration.The inhibition of tumor growth may pass to improve the tumorbearing mice with immune status and enhance the body's anti-tumor capacity.

The experimental research anti-tumor effect of modified decoction for driving out blood stasis in the blood mannsion / 国际中医中药杂志

Objective Modified Decoction for Driving Out Blood Stasis in the Blood Mannsion on ascites tumor model of transplanted H22 anti-tumor effect and the impact of VEGF. Methods Adult male mice 100, inoculated with H22 hepatoma cells, the establishment of ascites H22 transplanted tumor model, then divided into 10 groups were given saline, capecitabine, flavored Modified Decoction for Driving Out Blood Stasis in the Blood Mannsion (high, medium and low dose) was administered orally for 10 days, 11 days of treatment, observation of suppression tumor rate, the rate of change in life extension; detected by immunohistochemistry the expression of VEGF in tumors, SPSS13.0 software for statistical analysis. Results Capecitabine, flavored Xuefuzhuyutang (high, medium and low dose) inhibited tumor growth rates were 60%, 49%, 41%, 35%, life extension rate of the three groups were 1.68% 157.98%, 70.58%, 49.57% higher. Conclusion Modified Decoction for Driving Out Blood Stasis in the Blood Mannsion can inhibit tumor cell proliferation in tumor-bearing mice, significantly prolonged the survival time of mice, reduce the tumor tissue and tumor tissue expression of VEGF.