Anti-tumor Effect and Mechanism of Pratia Extract on H22 Tumor-bearing Mice

[Objectives]This study was conducted to investigate the inhibitory effect of pratia extract on H22 tumor-bearing mice and the effects on immune organs.[Methods]With the application of H22 liver tumor-bearing mice as an animal model,the animals were divided into such three Pratia extract groups as the high,medium and low dose groups(400,200 and 100 mg/kg)and cyclophosphamide CTX group(20 mg/kg).15 d after the administration,the animals were killed by cervical dislocation,and the tumors,thymuses and spleens were taken and weighed,followed by the calculation of the tumor inhibitory rate and the thymus and spleen index,and the serum tumor necrosis factor-α(TNF-α)and interleukin-2(IL-2)levels were determined by ELISA assay.[Results]The inhibitory rates were 54.1,32.6 and 8.2%,respectively,and there were significant differences from the model group(P<0.05);and the spleen index of the tumor-bearing mice was reduced,while the thymus index was improved.The serological results showed that the drug-administrated groups significantly improved the IL-2 levels in the tumor-bearing mice,but had no effects on TNF-α.[Conclusions]Pratia extract has an antitumor effect on H22 tumor-bearing mice,and show certain dose-effect relationship,and its mechanism may be related to enhancing the immune function in tumor-bearing mice by regulating IL-2.

粗毛纤孔菌与桑叶发酵产物——粗桑茶对H22荷瘤小鼠抗肿瘤作用

以桑叶为培养基质,利用粗毛纤孔菌(Inonotus hispidus)菌株进行发酵,制备粗桑茶,用粗桑茶(低、中、高剂量分别为195、390、780 mg·kg^(-1))灌胃H22荷瘤小鼠,测定小鼠体质量、肿瘤体积、肿瘤抑制率和胸腺、肝脏、肾脏、脾脏指数,检测血清中IL-2、IL-6、TNF-α、IFN-γ含量,用苏木素-伊红染色观察小鼠肝脏、肾脏、脾脏、肿瘤组织,用免疫组化染色观察小鼠肿瘤组织,并采用Western blot方法检测肿瘤相关蛋白表达情况。结果表明:与模型组相比,粗桑茶各剂量组肿瘤体积均极显著减小;粗桑茶高剂量组小鼠肿瘤质量极显著降低;粗桑茶高剂量组胸腺指数极显著升高;粗桑茶低剂量组小鼠血清中的IL-2、IL-6,中剂量组的TNF-α,高剂量组的IL-2含量显著降低;粗桑茶低剂量组的TNF-α、IFN-γ,中剂量组的IL-2、IFN-γ,高剂量组的IL-6、TNF-α、IFN-γ含量显著增加;粗桑茶各剂量组小鼠肿瘤组织的Bax、Cytc、Parp、TNF-α、caspase-3、cleaved caspase-3、caspase-9、 cleaved caspase-9蛋白表达量显著增加,Bcl-2蛋白表达量显著降低。研究结果表明粗桑茶具有抗肿瘤活性,说明采用粗毛纤孔菌菌株发酵桑叶制备产品的思路可行,为进一步工业化生产提供参考。

地塞米松对小鼠H_(22)肿瘤生长及血管内皮生长因子表达的影响

目的 探讨地塞米松对小鼠H2 2 肿瘤生长及血管内皮生长因子 (VEGF)表达的影响。方法 BALB c小鼠皮下接种H2 2 肿瘤细胞 ,腹腔注射地塞米松 ,每天测肿瘤直径。用抗CD31单抗对肿瘤组织做免疫组化微血管计数。噻唑蓝法检测地塞米松对体外培养的H2 2 细胞和人脐静脉内皮细胞ECV 30 4增殖的影响。RT PCR法测定肿瘤组织和体外培养细胞的VEGFmRNA表达。结果 地塞米松可以显著抑制体内H2 2 肿瘤的生长 ,给药组肿瘤体积与对照组比较P <0 .0 5。给药组微血管计数和VEGFmRNA表达比对照组显著减少。体外实验中 ,随着地塞米松浓度的增加 ,ECV 30 4细胞的增殖率降低 ;H2 2 细胞中VEGFmR NA表达下降。结论 地塞米松可以显著抑制小鼠H2 2 肿瘤的生长。地塞米松对体内H2 2 肿瘤组织和体外H2 2 细胞的VEGFmRNA表达均有显著的抑制作用。地塞米松对肿瘤细胞VEGF表达的抑制作用可能是其抗H2 2 肿瘤及抗血管生成的一个重要原因。

十全育真汤对H_(22)荷瘤小鼠免疫功能的影响

目的:研究十全育真汤对荷瘤小鼠免疫功能的影响,探讨十全育真汤抗肿瘤的作用机制。方法:SPF级雄性昆明种小鼠30只,制成荷H_(22)小鼠肝癌细胞移植瘤模型,随机分为模型组、阳性对照组及十全育真汤组(n=10);另选择未接种小鼠10只为正常对照组。正常对照组、模型组每天按10 ml/kg灌胃生理盐水及蒸馏水,阳性对照组、十全育真汤组每天按8 g/kg、18 g/kg灌胃参一药液(80 mg/ml)与十全育真汤剂。连续给药14 d后处死小鼠,测量胸腺、脾脏指数及抑瘤率,检测外周血中白细胞、淋巴细胞含量及T细胞亚群CD3、CD4、CD8细胞百分比,血清中白细胞介素2(IL-2)、肿瘤坏死因子α(TNF-α)及干扰素-β(IFN-β)的含量,淋巴细胞增殖能力及NK细胞杀伤功能。结果:较正常对照组,十全育真汤组小鼠体重明显增加,脾脏指数显著增大(P<0.05);白细胞、淋巴细胞CD4、CD8、CD3及TNF-α含量明显升高(P<0.05);IL-2、IFN-β含量显著下降(P<0.05);淋巴细胞增殖能力、NK细胞杀伤功能明显升高(P<0.05)。与模型组相比,十全育真汤组小鼠胸腺、脾脏指数显著增大(P<0.05);白细胞、淋巴细胞CD3、CD4、IL-2、IFN-β及TNF-α含量明显升高(P<0.05),CD8含量则显著降低(P<0.05);NK细胞杀伤功能及淋巴细胞增殖能力显著增加(P<0.05)。结论:十全育真汤可促进H_(22)荷瘤小鼠免疫器官的生长,增强机体免疫功能,有利于肿瘤机体的恢复。

Possible mechanisms associated with immune escape and apoptosis on anti-hepatocellular carcinoma effect of Mu Ji Fang granules

BACKGROUND Hepatocellular carcinoma(HCC)is one of the most common digestive system cancers with high mortality rates worldwide.The main ingredients in Mu Ji Fang Granules(MJF)are alkaloids,flavonoids,and polysaccharides.MJF has been used in the clinical treatment of hepatitis,cirrhosis and HCC for more than 30 years.Few previous studies have focused on the mechanism of MJF on tumor immunology in the treatment of HCC.AIM To explore the mechanism of action of MJF on tumor immunology in the treatment of HCC.METHODS The absorbable ingredients of MJF were identified using Molecule Network related to High Performance Liquid Chromatography-Electron Spray Ionization-Time of Flight-Mass Spectrometry,and hub potential anti-HCC targets were screened using network pharmacology and pathway enrichment analysis.Forty male mice were randomly divided into the Blank,Model,and MJF groups(1.8,5.4,and 10.8 g/kg/d)following 7 d of oral administration.Average body weight gain,spleen and thymus indices were calculated,tumor tissues were stained with hematoxylin and eosin,and Interferon gamma(IFN-γ),Tumor necrosis factorα(TNF-α),Interleukin-2,aspartate aminotransferase,alanine aminotransferase,alpha-fetoprotein(AFP),Fas,and FasL were measured by Enzyme-linked Immunosorbent Assay.Relevant mRNA expression of Bax and Bcl2 was evaluated by Real Time Quantitative PCR(RTqPCR)and protein expression of Transforming growth factorβ1(TGF-β1)and Mothers against decapentaplegic homolog(SMAD)4 was assessed by Western blotting.The HepG2 cell line was treated with 10 mg/mL,20 mg/mL,30 mg/mL,40 mg/mL of MJF,and another 3 groups were treated with TGF-β1 inhibitor(LY364947)and different doses of MJF.Relevant mRNA expression of TNF-α,IFN-γ,Bax and Bcl2 was evaluated by RT-qPCR and protein expression of TGF-β1,SMAD2,p-SMAD2,SMAD4,and SMAD7 was assessed by Western blotting.RESULTS It was shown that MJF improved body weight gain and tumor inhibition rate in H22 tumorbearing mice,protected immune organs and liver function,reduced the HCC indicator AFP,affected immunity and apoptosis,and up-regulated the TGF-β1/SMAD signaling pathway,by increasing the relative expression of TGF-β1,SMAD2,p-SMAD2 and SMAD4 and decreasing SMAD7,reducing immune factors TNF-αand IFN-γ,decreasing apoptosis cytokines Fas,FasL and Bcl2/Bax,and inhibiting the effect of LY364947 in HepG2 cells.CONCLUSION MJF inhibits HCC by activating the TGF-β1/SMAD signaling pathway,and affecting immune and apoptotic cytokines,which may be due to MJF adjusting immune escape and apoptosis.

Effect of Qinggan Huayu granule on H22 liver ascitic tumor mice

Objective:To evaluate the therapeutic effect of Qinggan Huayu granule on mice with H22 liver cancer ascites tumor.Methods:A H22 liver cancer ascites mouse model was established by intraperitoneally injecting H22 liver cancer cells.Mice were randomly divided into the model group,the Ganfule group(1.35 g/kg),the fluorouracil group(50 mg/kg i.p),the Qinggan Huayu granule groups at low(0.67 g/kg),medium(1.34 g/kg),and high(2.68 g/kg)doses.Then the mice were administered continuously for 10 days and body weight and abdominal circumference were monitored every 3 days.On day 11,eight rats in each group were randomly selected for dissection to detect the amount of peritoneal water,peritoneal permeability and histopathological changes.The remaining mice were observed for survival.In addition,the vascular endothelial growth factor A(VEGFA)and vascular endothelial growth factor receptor 2(VEGFR2)were determined by Western blotting.Results:Compared with the model group,the weight growth of mice in the fluorouracil group and the medium-dose and high-dose Qinggan Huayu granule groups was slower(P<0.05).Moreover,the abdominal circumference of mice in each treatment group was increased slowly.There were significant differences in abdominal circumference between the fluorouracil group,the medium-dose group and the control group from day 6(P<0.05)while the abdominal circumference of the high dose group was significantly smaller than that of the control group from day 12(P<0.05).Moreover,compared with the model group,the amount of ascites in the medium-and high-dose Qinggan Huayu granule groups was decreased significantly(P<0.05).The optical density value of ascites supernatant in medium-and high-dose Qinggan Huayu granule group and the fluorouracil group decreased significantly(P<0.05)and the survival period of the medium-dose Qinggan Huayu granule group and the fluorouracil group was prolonged prominently(P<0.05).There was no significant difference in the low-dose Qinggan Huayu granule group and the Ganfule group.Peritoneal histopathological assay showed more complete peritoneal structure,less edema and less angiogenesis of the peritoneum in the fluorouracil group and the medium-and high-dose Qinggan Huayu granule group,which was better than that of the Ganfule group and the low-dose group.Compared with the model group,the expressions of VEGFA and VEGFR2 in the medium-dose Qinggan Huayu granule group decreased significantly(P<0.05,P<0.01).Conclusion:Qinggan Huayu granule can inhibit ascites production in the mice model with H22 liver cancer ascites tumor,prolong the survival of mice,and reduce peritoneal permeability and suppress the increase of peritoneal neovascularization.The mechanism may be related to the inhibition of VEGF/VEGFR pathway.

叶下珠复方Ⅱ号对小鼠H22肝癌移植瘤生长的抑制作用

目的观察叶下珠复方Ⅱ号对小鼠H22肝癌移植瘤生长的抑制作用,并探讨其可能的作用机制。方法取昆明小鼠40只,建立小鼠H22肝癌移植瘤模型,分为模型组、叶下珠复方Ⅱ号高剂量组(37.5g/kg)、低剂量组(18.75g/kg)和环磷酰胺(20mg/kg)治疗组。叶下珠复方Ⅱ号高、低剂量组均灌胃给药,环磷酰胺组采用腹腔注射,每日1次,连续8d。末次给药24h后,测定瘤体质量,计算抑瘤率;检测血清IL-2和TNF-а含量。结果叶下珠复方Ⅱ号高、低剂量组瘤重明显低于模型组,差异均有显著性意义(P<0.01),但与环磷酰胺组比较,差异均无显著性意义(P>0.05)。叶下珠复方Ⅱ号高、低剂量组的抑瘤率分别为67%和58%,与环磷酰胺组接近。叶下珠复方Ⅱ号高剂量组血清IL-2水平明显高于模型组(P<0.05)。叶下珠复方Ⅱ号高、低剂量组血清TNF-а水平明显低于模型组,差异有显著性意义(P<0.05)。结论叶下珠复方Ⅱ号有较好的抑制小鼠H22肝癌移植瘤生长作用,作用机制与调节免疫,升高IL-2水平和抑制TNF-а水平有关。

凯息平口服液对H_(22)荷瘤小鼠的抑瘤生长作用及抑瘤机理探讨

凯息平口服液对小鼠移植性H2 2 实体瘤有明显的抑瘤生长作用 ,2 7.0、18.0、9.0g/kg剂量的抑瘤率分别 4 7.6 %、4 7.8%和 33.8% ;对H2 2 腹水癌小鼠有一定的延长生存期的作用 ,强于阳性对照药复方天仙胶囊 ,且没有氟尿嘧啶 (5 FU)所表现的毒副作用。对体外培养的H2 2 细胞有一定程度的抑制作用 ,最低有效药物浓度可达 2 5mg/L ,抑制作用可维持 4 8h以上。但对培养的脾T细胞 ,其剂量达 80 0mg/L时才表现出抑制增殖的作用。表明凯息平口服液对肿瘤细胞有独特的抑制作用 ,绝非细胞毒所致。实验还表明 :凯息平口服液能增强荷瘤鼠的T、B淋巴细胞增殖能力 ;对自然杀伤细胞 (NK)的活性也有明显的促进作用 ,同时明显增强腹腔巨噬细胞 (MΦ)的吞噬活性 ,显示了其在抗肿瘤免疫中具有重要的作用。

Anticancer Effects of Zaoxiu Compound Decoction on H_22 Mice with Heps

Objective To investigate the anticancer effects of the Zaoxiu Compound Decoction(ZCD) on H22 mice with Heps and the influence on the construction of H22 cells. Methods With in vivo techniques, the antineoplastic drugs were given to hepatoma H22 model mice which were randomly divided into negative control(physiological saline), positive control(cyclophosphamide), high- and low-dose ZCD groups. After 10-d administration, the pathological examinations of tumor tissue, livers, and kidneys in mice were carried out and the inhibitory rate on tumor was calculated. Results The high-and low-dose ZCD had obvious inhibition on H22 cells. The pathological observation showed that high- and low-dose ZCD had a strong inhibitory effect on H22 cells as well as positive control, with few toxic effects to the livers and kidneys in mice. Conclusion ZCD has the anticancer effect and has no obvious toxic effects on the model mice.