Hu-Qi-Zheng-Xiao Decoction Inhibits the Metastasis of Hepatocellular Carcinoma Cells by Suppressing the HIF-1α Signaling Pathway to Inhibit EMT, LCSC, and Angiogenic Process.

BACKGROUND: Hepatocellular carcinoma (HCC) is a common clinical malignant tumor of the digestive system. Hu-Qi-Zheng-Xiao (HQZX) decoction has been clinically found to prolong the survival of patients with hepatocellular carcinoma and improve the quality of patients' survival, but its antitumor biological mechanism is still unclear. METHODS: A nude mouse hollow fiber hepatocellular carcinoma model was constructed to analyze the in vivo efficacy of HQZX decoction against 7 different hepatocellular carcinoma cells. The subcutaneous graft tumor model was again validated. In vitro, the effect of HQZX decoction on the growth and metastasis of the cell line with the highest growth inhibition was evaluated. The cell line with the best efficacy response screened was again used to construct a hollow fiber hepatocellular carcinoma model and hollow fiber conduit cells were extracted to detect the expression of HIF-1α, VEGF, EMT-related molecules, LCSCs-related molecules, and to observe the density of the subcutaneous vascular network of hollow fiber conduits. The liver metastasis model of splenic injection was constructed to observe the effect of HQZX decoction on tumor metastasis. RESULTS: The hollow fiber hepatocellular carcinoma model was evaluated for the efficacy of HQZX decoction, and it was found to have the highest growth inhibition of LM3-luc cells. In vitro, the CCK8 assay revealed that HQZX decoction could inhibit tumor migration and invasion and promote apoptosis. In addition, the mechanism study of extracting cells from hollow fiber tubes found that HQZX decoction could inhibit metastasis-associated HIF-1α, VEGF, EMT-related molecules, and LCSCs-related molecules expression. capillary network around subcutaneous fiber tubes was reduced in the HQZX decoction gavage group of mice. It inhibited tumor metastasis in nude mice. CONCLUSIONS: HQZX decoction inhibited the growth of a variety of hepatocellular carcinoma cells. HQZX decoction suppressed the expression of metastasis-associated VEGF, EMT-related molecules, and LCSCs-related molecules and inhibited tumor angiogenesis and growth and metastasis, which may be related to the inhibition of the HIF-1α signaling pathway. It reveals that HQZX decoction may be a promising herbal compound for anti-HCC therapy, and also reveals the accurate feasibility of the hollow fiber hepatocellular carcinoma model for in vivo pharmacodynamic evaluation and mechanism study.

Response of soil stoichiometric characteristics to climatic factors in temperate steppe of Longzhong region, China.

We explored the relationship between climate factors (mean annual precipitation and mean annual temperature) and the contents and stoichiometry of soil carbon (C), nitrogen (N), and phosphorus (P) at different soil depths (0-5, 5-10, 10-20, 20-30, 30-50, 50-70, and 70-100 cm) temperate steppe of Longzhong. The results showed with the increases of soil depth, soil C, N contents, C:P, and N:P gradually decreased from 21.88 g·kg-1, 1.84 g·kg-1, 33.6 and 3.1 to 7.67 g·kg-1, 0.59 g·kg-1, 12.5 and 1.0, respectively. Soil C:N showed an increasing trend from 12.2 to 13.9, while soil P content remained stable with an average of 0.61 g·kg-1. Soil C, N, C:P, and N:P were significantly positively correlated with mean annual precipitation and negatively correlated with mean annual temperature. Soil P content and C:N were not correlated with mean annual precipita-tion and mean annual temperature. With the increases of soil depth, the total explanatory power of the changes in soil C, N and P contents by mean annual precipitation and mean annual temperature decreased and then increased, and that in soil C:P, N:P and C:N did not change significantly. The changes of soil C, N and P contents on the temperature steppe were mainly influenced by mean annual precipitation. The effects and relative contributions of mean annual precipitation and mean annual temperature on the variations of soil nutrient contents and stoichiometry of C, N and P differed at different soil depths.

Pien Tze Huang accelerated wound healing by inhibition of abnormal fibroblast apoptosis in Streptozotocin induced diabetic mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Diabetic foot ulcer is one of the most serious complications of diabetes. Effective medical treatment regarding improvement of ulcer healing in patients is essential. Pien Tze Huang (PZH), a valuable Chinese traditional medicine, has been found significant efficacy on the curing of diabetic wound in clinic recently. AIM OF THE STUDY: This work was conducted to confirm the efficacy, and compare the therapeutic effect through the oral administration and local delivery route, providing a rationale for the new PZH form development; besides, the mechanisms through which PZH promoted the wound healing was also discussed. MATERIALS AND METHODS: First, the chemical composition of PZH was characterized by 1H-NMR and HPLC. The anti-apoptosis effects of PZH on high concentration glucose injured epidermal fibroblast (HFF-1) was investigated in a dose dependent way. Then, the effects of the systematical administration of PZH, and the topical used route on excisional wounds of Streptozotocin (STZ) induced diabetic mice were compared. RESULTS: The results illustrated that PZH decreased the reactive oxygen species (ROS) levels in cells, preventing cell damage/apoptosis through an ROS/Bcl-2/Bax/Caspase-3 pathway. The in vivo study proved that topical use of PZH exceeded the systematical route both in accelerating the wound closure and improving the healing quality. Meanwhile, PZH promoted wound closure through stimulating the secretion of Col-I, decreasing fibroblast apoptosis, and enhancing myo-fibroblast differentiation, in consistent with the mechanism study in vitro. CONCLUSIONS: Local used PZH improves wound healing by inhibiting the abnormal HFF-1 apoptosis and senescence. The study held a great promise for development of a topical dosage form of PZH for diabetic wound healing.

ASPP2 enhances chemotherapeutic sensitivity through the down-regulation of XIAP expression in a p53 independent manner in hepatocellular carcinoma.

Apoptosis stimulated protein of p53-2 (ASPP2) induces the transcription of p53-targeted genes to stimulates its pro-apoptosis function. The poor chemotherapeutic sensitivity is associated with the decreased ASPP2 expression in many human cancers. Here, multiple genes real-time RT-PCR array and western blotting analysis show that ASPP2 suppress the expression of X-linked inhibitor of apoptosis protein (XIAP), determinant of chemoresistance in cancer, in hepatocellular carcinoma (HCC) in a p53-independent manner. Further experiments with ASPP2-rAd and ASPP2-Lv confirmed that ASPP2 enhanced sensitivity of sorafenib to HCC via suppressing XIAP expression. XIAP mainly found on the cytoplasm and perinuclear areas of ASPP2 over-expressed HepG2 cells, while both cytoplasm and nucleus in ASPP2 shut down HepG2 cells. The association of poor sensitivity of sorafenib and XIAP expression was also found both in ASPP2 shut down and overexpress mice, where liver tissue with decreased or increased ASPP2 displayed less or more apoptosis, respectively. Finally, ASPP2 and XIAP expression analyzed in 43 hepatocellular carcinoma tumors and 44 adjacent normal tissues from 38 hepatocellular carcinoma patients for fully understand their expression within HCC patients. Compared with the tumor tissues, ASPP2 mRNA levels were increased, and XIAP levels decreased in the adjacent normal tissues. Taken together, XIAP suppressed ASPP2 increased tumor sensitivity to chemotherapy in a p53-independent manner, which was associated with chemotherapy resistance, suggesting that p53 activation and XIAP suppression were two independent ways that ASPP2 enhance the sensitivity of chemotherapy.

Antitumor Efficacy of Huqizhengxiao (HQZX) Decoction Based on Inhibition of Telomerase Activity in Nude Mice of Hepatocarcinoma Xenograft.

OBJECTIVE: Huqizhengxiao (HQZX) decoction is a mixture of traditional Chinese medicines comprising 10 herbs, with inhibitory effects on hepatocarcinoma. The aim of the study is to observe the antitumor efficacy and mechanism of HQZX decoction in nude mice with hepatocellular carcinoma xenografts. METHODS: HepG2-luc subcutaneous hepatocarcinoma was established in nude mice. The mice were divided into 5 groups: control, cinobufagin, HQZXS, HQZXM, and HQZXH with doses 13.52, 27.03, and 54.06 g/kg, respectively. HQZX decoction was prepared for intraperitoneal intragastric administration for 3 weeks. Tumor growth was measured with Vernier calipers and in vivo imaging system. α-Fetoprotein (AFP) was determined by radioimmunoassay. Tumor necrosis factor-α (TNF-α) was measured with enzyme-linked immunosorbent assay (ELISA) assay. Telomerase activity was measured with polymerase chain reaction-ELISA. Nuclear mitosis and necrosis were observed with hematoxylin-eosin stain. Apoptotic proteins of caspase-3, Bcl-2, and Bax were examined by Western blot. Signaling molecules of ERK, mTOR, and STAT3 were measured with Luminex assay. RESULTS: HQZX decoction showed good inhibition of HepG2-luc xenografts. Compared with control group, the relative tumor proliferation rate was less than 60% in the HQZXH and HQZXS. The tumor inhibition rate of HQZXH group reached 52% ± 15%. Relative average optical density values of the HQZXS and HQZXH groups decreased significantly. The mitotic index in HQZXS, HQZXM, and HQZXH groups decreased greatly. Telomerase activity of HQZXS was clearly reduced, and, the caspase-3 expression upregulated in HQZXH group. Bcl-2 expression was downregulated in HQZXS and HQZXH. The ratios of p-ERK/ERK and p-STAT3/STAT3 in HQZXS group were significantly downregulated. CONCLUSION: HQZX decoction can clearly inhibit the growth of hepatocellular carcinoma and induce tumor apoptosis. Its antitumor mechanism may be related to reducing telomerase activity and regulating the STAT3 and ERK signal pathway.

A retrospective study of supplemental iron intake in singleton pregnancy women with risk of developing gestational diabetes mellitus.

This study investigated the effect of supplemental iron intake (SII) in early singleton pregnancy women with the risk of developing gestational diabetes mellitus (GDM) among Chinese population.This study included 259 singleton pregnancy participants. Of those, 135 women underwent SII and were assigned to an intervention group, while 124 participants received no SII and were assigned to a control group. The outcome measurements consisted of the number of patients with GDM development, the levels of hemoglobin (Hb) and ferritin, and the outcomes of infant at delivery.No significant difference in the number of patients with GDM development was found between 2 groups at delivery. However, when compared with control group, subjects in the intervention group showed greater efficacy in delivery mode choice of vaginal delivery (P = .04), and cesarean section (P = .01), as well as the birthweight of infants (P < .01). Moreover, Hb and ferritin levels were also significantly higher in the intervention group than those in the control group (P < .01).The results of this retrospective study showed that SII may not increase risk of developing GDM in singleton pregnancy women; and also may benefit both pregnancy women and infants among Chinese population.

Extract of Stellerachamaejasme L(ESC) inhibits growth and metastasis of human hepatocellular carcinoma via regulating microRNA expression.

BACKGROUND: MicroRNAs(miRNAs)are involved in the initiation and progression of hepatocellular carcinoma. ESC, an extract of Stellerachamaejasme L, had been confirmed as a potential anti-tumor extract of Traditional Chinese Medicine. In light of the important role of miRNAs in hepatocellular carcinoma, we questioned whether the inhibitory effects of ESC on hepatocellular carcinoma (HCC) were associated with miRNAs. METHODS: The proliferation inhibition of ESC on HCC cells was measured with MTT assay. The migration inhibition of ESC on HCC cells was measured with transwell assay. The influences of ESC on growth and metastasis inhibition were evaluated with xenograft tumor model of HCC. Protein expressions were measured with western blot and immunofluorescence methods and miRNA profiles were detected with miRNA array. Differential miRNA and target mRNAs were verified with real-time PCR. RESULTS: The results showed that ESC could inhibit proliferation and epithelial mesenchymal transition (EMT) in HCC cells in vitro and tumor growth and metastasis in xenograft models in vivo. miRNA array results showed that 69 differential miRNAs in total of 429 ones were obtained in MHCC97H cells treated by ESC. hsa-miR-107, hsa-miR-638, hsa-miR-106b-5p were selected to be validated with real-time PCR method in HepG2 and MHCC97H cells. Expressions of hsa-miR-107 and hsa-miR-638 increased obviously in HCC cells treated by ESC. Target genes of three miRNAs were also validated with real-time PCR. Interestingly, only target genes of hsa-miR-107 changed greatly. ESC downregulated the MCL1, SALL4 and BCL2 gene expressions significantly but did not influence the expression of CACNA2D1. CONCLUSION: The findings suggested ESC regressed growth and metastasis of human hepatocellular carcinoma via regulating microRNAs expression and their corresponding target genes.

Anti-tumor pharmacological evaluation of extracts from stellera chamaejasme L based on hollow fiber assay.

BACKGROUND: Stellera chamaejasme L, a traditional Chinese herb, has long been used for treatment of various tumors in the Chinese population. In our previous study, we paid an attention to the cytotoxic and proapoptotic effects of Stellera chamaejasme L extracts (ESC, ESC-1 and ESC-2, the latter two were isolated from ESC) on 4 various tumor cells (NCI-H157, NCI-H460, BEL-7402 and SK-HEP-1) in vitro. ESCs showed significantly inhibitory effects on the 4 tumor cells. ESC-2 had the strongest inhibitory effect and the broadest sensitive cell spectrum. ESC-2 and ESC acted in a similar way against tumor cells, which suggested anti-tumor active fraction of ESC might exist in ESC-2. Here, we further observe the inhibitory and proapoptotic effects of Stellera chamaejasme L extracts in vivo. METHODS: In this study, we used hollow fiber tumor model to evaluate the inhibitory and proapoptotic effects of Stellera chamaejasme L extracts. Apoptotic rates of the cancer cells retrieved from the hollow fibers were measured with flow cytometric analysis, caspase 3, 8, 9 enzyme activities were detected by colorimetric assay, Fas, Fas-L, TNF-R1 and TNF-α expression were determined with elisa assay and radioimmunoassay respectively. RESULTS: The results showed that ESC, ESC-2 all had inhibitory effects on 4 tumor cells. According to the effect strength, dose and antitumor spectrum, the order of antitumor effects of ESCs was: ESC-2 > ESC > ESC-1. NCI-H460 cells were the most sensitive to ESCs. ESC, ESC-2 increased greatly the apoptotic rate and caspase 3, 8 enzyme activities in NCI-H460. ESCs had no significant effects on expression of Fas, Fas-L protein, but TNF-α/TNFR1 protein expression in NCI-H460 cells changed significantly after ESC and ESC-2 treatment. CONCLUSION: ESC-2 had the similar antitumor effect to that of ESC in vivo and further confirmed that ESC-2 may be the main antitumor active fraction of ESC, which was consistent with our previous results in vitro.

Stellera chamaejasme L. extract induces apoptosis of human lung cancer cells via activation of the death receptor-dependent pathway.

Stellera chamaejasme L. has been widely used in the treatment of lung, liver and esophageal cancer in Chinese traditional medicine. In our previous study, we found that the extract of Stellera chamaejasme L. (ESC) inhibited the growth and induced the apoptosis of human lung cancer NCI-H157 cells. In the present study, we investigated the cellular effects of an ESC-2 extract isolated from ESC in the NCI-H157 human lung cancer cell line. We found that ESC-2 inhibited the growth of NCI-H157 cells, demonstrating ESC-2-induced morphological changes in cells and reduced cell viability in a dose- and time-dependent manner. Furthermore, we observed that ESC-2 resulted in apoptosis, activation of caspase-8 and caspase-3, and an increase in Fas expression, indicating that the NCI-H157 cell growth inhibitory activity of ESC-2 was due to death receptor-dependent pathway-mediated apoptosis, which may partly explain the anti-cancer activity of ESC-2.

[Comparative study on tumor cell apoptosis in vitro induced by extracts of Stellera chamaejasme].

OBJECTIVE: To in vitro compare the induction of extracts of Stellera chamaejasme ESC, ESC-1 and ESC-2 on NCI-H157 cell apoptotic. METHOD: The apoptosis rate was inspected by flow cytometry; caspase-3, 8, 9 activities was measured by spectrophotometry. Fas, Fas-L, TNF-alpha, Trail-R, Cyto-C, Smac/diablo protein expressions of apoptosis pathway was observed by Elisa method. RESULT: Compared with the control group, ESC, ESC-1, ESC-2 can significantly improve the apoptosis rate of NCI-H157 cell. ESC significantly improved cells caspase-3, 8 activity, ESC-2 can significantly improve the activity of caspase-3, 8, 9. ESC, ESC-1, ESC-2 significantly increased Fas expression and ESC significantly increased Fas/Fas-L ratio. ESC, ESC-1, ESC-2 significantly increased TNF-alpha protein expression. ESC-1 significantly lowered TRAIL-R expression. ESC, ESC-1, ESC-2 had no significant effect on Cyto-C. ESC-1, ESC-2 significantly reduced Smac protein expression. CONCLUSION: The apoptotic effect induced by ESCs may be related to the regulation of death receptor pathway proteins. Induction mechanisms of ESCs were so complicated that it may have a two-way regulatory effect. Its induction in apoptosis is a result from comprehensive regulation and control.

[Study on impact of vinegar processing on toxic effect of Stellera chamaejasme].

OBJECTIVE: To study the scientific theory on detoxification (attenuation) of Stellera chamaejasme (ScL) by processing and the impact on drug effect of ScL before and after being processed with vinegar. METHOD: The difference in ingredients of ScL before and after being processed vinegar was compared by using PHLC-MS technique. A subcutaneously transplanted tumor model of H22 hepatoma was established to compare the lethal effect and weight change between tumor-loaded mice and normal mice. After consecutive oral administration in tumor-loaded mice, the impacts on tumors and immune organs were compared before and after being processed with vinegar. Luciferase Report Gene was employed to investigate the target genes TGF-beta, AP1 and NF-kappaB. RESULT: The LD50 (median fatal dose) of extract Zp1102 exhibited higher than that of the processed one Zp1103, that is 9. 89 g x kg(-1) vs. 16.85 g x kg(-1). According to the test, Zp1102 showed more effective anti-tumor activities in vivo than that of Zp1103 in a same dosage, with the tumor inhibitory rate 36.24% (P < 0.01) at the dosage of2 g x kg(-1) and 34.40% (P < 0.05) at the dosage of 1 g x kg(-1). At the dosage of 1 g x kg(-1), Zp1102 showed a tumor inhibitory rate of 34.52% (P < 0.05), much higher from 21.55% in Zp1103. Both Zp1102 and Zp1103 had basically no impact on the report gene NF-kappaB, besides that Zp1102 up-regulated the report gene after increase in NF-kappaB concentration and down-regulated TGF-beta, but Zp1103 can only up-regulate NF-kappaB expression without any impact on TGF-beta. CONCLUSION: Processed ScL extracts show less toxic than unprocessed extracts and slight reduction in anti-tumor activity, which may be related to the regulation of transforming growth factor TGF-beta.

[Regularity analysis on clinical treatment in primary liver cancer by traditional Chinese medicine].

OBJECTIVE: To evaluate clinical treatment regularity of traditional Chinese medicine (TCM) on primary liver cancer and provide inspiration for the clinical use. METHOD: Traditional Chinese medicine database on primary liver cancer was established to analysis the classification, frequency, dosage of TCM in clinical treatment. RESULT: Tonic medicine is the most common medication, herbs for heat-clearing, promoting blood circulation for removing blood stasis, eliminating dampness and diuresis and regulating flow of Qi are more common medication, herbs for relieving exterior disorder and digesting are common medication; the first frequency of single herb is Atractglodis Macrocephalae Rhizoma, Poria, Codonopsis Radix. Popular classical prescriptions are Sijunzi Tang, Xiaochaihu Tang, YiguanJian, Xiangsha Liujunzi Tang, Xiaoyao Wan and Gexia Zhuyu Tang, Liuwei Dihuang Tang and Yinchenhao Tang et al. Gallic Gigerii Endothelium Corneum and Ophiopogonis Radix are most commonly drug for poor appetite. Astragali Radix is most commonly drug for fatigue. Corydalis Rhizoma, Toosendan Fructus are most common for liver pain; Pericarpium Arecae, Polyporus, Poria are most common herbs for ascites; Artemisiae Scopariae Herba is common drug for jaundice. CONCLUSION: Replenishing qi to invigorate the spleen, sparsing liver to regulate the flow of vital energy, clearing heat and promoting diuresis, promoting blood circulation for removing blood stasis, nourishing yin and detoxification are the main principles for treating primary liver cancer. Improving clinical symptoms, signs and quality of life of patients with TCM is the key to clinical treatment.

In vitro inhibitory and pro-apoptotic effect of Stellera chamaejasme L extract on human lung cancer cell line NCI-H157.

OBJECTIVE: To investigate the inhibitory and pro-apoptotic effect of Stellera Chamaejasme L extract (ESC) in vitro. METHODS: ESC was first extracted with ethanol, and then washed using a polyamide column with 60% ethanol. ESC was then decompressively recycled and vacuum dried at room temperature to obtain active fractions. Subsequently, the cytotoxic and apoptotic effects of ESC on NCI-H157 human lung cancer cells were determined. RESULTS: The results showed that ESC was rich in isomers of Chamaejasminor, neochamaejasmine and Sikokianin. ESC had significant cytotoxicity against NCI-H157 cells, with an IC50 of approximately 18.50 microg x mL(-). ESC caused significant increase in total apoptotic rate, the activity of caspase 3 and 8, CONCLUSION: The inhibitory effect of ESC on NCI-H157 tumor cells might partly be attributed to its apoptotic induction through activation of the Fas death receptor pathway.

[Influence of Tiangou Jiangya capsule on blood pressure in renovascular hypertension rats].

OBJECTIVE: To observe the effect of Tiangou Jiangya capsule (TJC) on blood pressure in renovascular hypertension rats and explore its possible mechanism. METHOD: Seventy-two Wistar rats were randomly divided into normal control group, model group, captopril group, TJC small, medium and high dose groups. Non-invasive blood pressure measurement was used to detect the arterial blood pressure of rat tails. PRA, Ang II , ALD, 6-Keto-PGF1alpha, ET and TXB2 content in blood was measured by radioimmunoassay. NO content in blood was determined by method of nitrate reductase. RESULT: The systolic, diastolic and mean pressure significantly increased, serum PRA, Ang II , ALD decreased, ET levels significantly increased in model group rats. TJC significantly reduced blood pressure, improved the plasma renin activity, decreased ET levels and increased NO content of model rats. CONCLUSION: TJC can reduce blood pressure of renovascular hypertention rats, and the mechanism may be related to its regulating lower blood pressure regulation of the secretion of RAAS system and improving vascular endothelial function.

[Effects of Wuji pill compound with different compatibility on cytochrome P450 CYP3A1/3A2 in rat liver microsomes in vitro].

OBJECTIVE: Wuji pill is a prescription of traditional Chinese medicine(TCM) and was composed of Rhizoma Coptidis, Fructus Evodiae Rutaecarpae and Radix Paeoniae Alba. The aim of this research is to investigate the effects of Wuji pill compound with different compatibility on the levels of enzymic activity of cytochrome P450 CYP3A1/3A2 in rat liver microsomes in vitro, and to confirm the compatibility mechanism of Wuji pill from the point of relationships between compound prescription of TCM and metabolism. METHOD: With testosterone being a probe, the levels of enzymic activity of CYP3A1/3A2 were detected by HPLC, which were suppressed by Wuji pill with different compatibility in vitro. RESULT: The IC50 of Rhizoma Coptidis, Fructus Evodiae Rutaecarpae, Radix Paeoniae Alba and 1"-9" of different level Wuji pill is: 38.96, 871.96, 15 519.17, 43.17, 60. 47, 276.12, 133.40, 118.08, 88. 47, 64. 36, 35. 13 and 39. 91 mg x L -', respectively. Rhizoma Coptidis and 1-9" of Wuji pill can suppress the enzymic activity of CYP3A1/3A2 significantly, and the capability of Rhizoma Coptidis in Wuji pill of action on CYP3A1/3A2 can be modified by different composition of Fructus Evodiae Rutaecarpae and Radix Paeoniae in Wuji pill, and there are statistical differences among the IC50 of 1#-9# of Wuji pill. While the ratio of Rhizoma Coptidis raises up in Wuji pill, Wuji pill may suppress the enzymic activity of CYP1A2 largely. CONCLUSION: The reason why Wuji pill with different compatibility has different pharmacodynamics and pharmacokinetics characteristics is likely to lie in the difference of the capability of Wuji pill with different compatibility on CYP3A1/3A2. [Key words] Wuji pill; CYP3A1/3A2; testosterone; HPLC; different compatibility prescription of traditional Chinese medi-cine

[Extracts of danshen and chuanxinlian on human monocytic line THP-1 induced by TNF-alpha].

OBJECTIVE: To investigate whether extracts of Danshen and Chuanxinlian (SL) could promote the function recovery in pre-monocytic cell line THP-1 induced by TNF-alpha. METHOD: SL extracts (0.125-2 g x L(-1)) were used to incubate THP-1 for 24 h before stimulation with TNF-alpha (20 microg x L(-1)), the adhesion, migration, lipid uptake and secretion of THP-1 were observed. RESULT: SL (0.5-2 g x L(-1)) had obvious effect on decreasing the THP-1 adhesion. The number of passed membrane was much fewer than that of control cells in SL (0.125-2 g x L(-1)). SL (0.125-2 g x L(-1)) reduced the total cholesterol content significantly. The levels of IL-6 in SL (2 g x L(-1)) were significantly decreased,and IL-10 was increased than that before the treatment. CONCLUSION: SL extracts could promote the function recovery such as adhesion, migration, lipid uptake and secretion of THP-1 induced by TNF-alpha, which probably is one of the mechanisms of inhibit the inflammatory reaction in initiation and development of AS.

[Anti-tumor effect of alcohol extract of Stellera chamaejasme in vitro].

OBJECTIVE: To observe the anti-tumor effect of alcohol extract of Stellera chamaejasme-AESC, AESC-1 and AESC-2 in vitro. METHOD: Inhibition rates of AESC, AESC-1, AESC-2 on five tumor cells (A549, NCI-H157, NCI-H460, BEL-7402 and SK-HEP-1 cell lines) were observed and IC50, GI50, TGI50 and LC50 were calculated with the methods of MTT and SRB. RESULT: AESC, AESC-1, AESC-2 have inhibitory effect on all of 5 cell lines and have a certain time-dependent nature. On 72 hours, inhibition rates of 100, 200 mg x L(-1) of various extracts on each cell line (except A549) were between 64.82% and 92.27%. IC50 of AESC and AESC-2 on NCI-H460 and BEL-7402 cell lines is less than 20 mg x L(-1), IC50 of AESC-1 on SK-HEP-1 cell line is less than 20 mg x L(-1). The mean GI50, TGI and LC50 of AESC, AESC-1, AESC-2 on sensitive cell lines were less than 40, 90, 150 mg x L(-1) respectively. CONCLUSION: AESC, AESC-1 and AESC-2 all have significant anti-tumor effect and have the different sensitivity to different cell lines; AESC-2 has the similar anti-tumor manner to the AESC and the anti-tumor active fraction of AESC may be mainly present in the AESC-2.

Inhibitory effect of schisandrin B on gastric cancer cells in vitro.

AIM: To investigate the inhibitory effect and possible mechanism of action of schisandrin B in SC-B on gastric cancer cells in vitro. METHODS: SC-B consisted of schisandrin B, aloe-emodin, and Astragalus polysaccharides. Exponentially growing human gastric cancer SGC-7901 cells were divided into six treatment groups: (1) control group (RPMI 1640 medium); (2) negative control group (2% DMSO); (3) positive control group (50 mg/L 5-Fluorouracil, 5-FU); (4) low-dose group (LSC, final concentration of schisandrin B, 25 mg/L); (5) moderate-dose group (MSC, final concentration of schisandrin B, 50 mg/L); (6) high-dose group (HSC, final concentration of schisandrin B, 100 mg/L). Follow-up was done at 12-48 h. An MTT (Methylthiazolyldiphenyl-tetrazolium bromide) assay was used to examine the inhibitory effect of SC-B on gastric cancer cells. The mitosis index was assessed using an inverted microscope. Flow cytometry was used to visualize the cell cycle. An RT-PCR (Reverse transcription-Polymerase chain reaction) -based assay was used to detect mRNA expression for cyclin D1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). RESULTS: The MTT assay showed that the number of living cells in the LSC, MSC and HSC groups was significantly smaller than that in the DMSO-treated group (P < 0.05) at 12-48 h. The inhibitory rate (IR) of the LSC group was 41.15% +/- 3.86%, 59.24% +/- 5.34% and 69.93% +/- 7.81% at 12, 24 and 48 h, respectively. The IR of the MSC group was 42.82% +/- 4.94%, 62.68% +/- 7.58% and 71.79% +/- 8.12% at 12, 24 and 48 h, respectively. The IR of the HSC group was 37.50% +/- 3.21%, 40.34% +/- 2.98% and 61.99% +/- 4.88% at 12, 24 and 48 h, respectively. These results suggested that a moderate dosage had the most obvious inhibitory efficacy at 48 h. Compared to the DMSO group, the mitosis index of the LSC, MSC, HSC groups was greatly decreased (P < 0.05) at all time points. Any dose of SC-B suppressed mitosis within 12-48 h. Compared to the DMSO group, the percentage of cells in the G0/G1 phase of the MSC group was greatly increased, and that of the S + G2M phase was greatly decreased, while the percentage of cell inhibition (PCI) in the MSC group was greatly increased (P < 0.05). This suggested that SC-B could exclusively arrest cells in the G0/G1 phase. Cyclin D1 mRNA expression was lower in the MSC group than that in the DMSO group (P < 0.05). CONCLUSION: SC-B can inhibit the proliferation and aberrant mitosis of human gastric cancer SCG-7901 cells in vitro. This inhibitory effect may be due to the down-regulation of cyclin D1 mRNA expression, which causes cell cycle arrest of gastric cancer cells.