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To detect the inhibitory effect of Astragalus protein on the proliferation of hepatocellular carcinoma cell line HepG2, transcriptomics was used to explore the anti-tumor mechanism of Astragalus protein. The dried roots of Astragalus was precipitated by ammonium sulfate to obtain Huang Qi protein (HQP) with different molecular weights. The effect of HQP on HepG2 and its toxic effect were detected by hemocytometry. Cell necrosis was detected by flow cytometry and Hoechst/propidium iodide (PI) double staining. The necrotic marker protein receptor interacting serine/threonine kinase 1 (RIP1) was determined by Western blot. Transcriptome sequencing was performed on the control group and dosing group RNA, and differential expression genes were analyzed for RNA-seq results. qRT-PCR was used to verified the relative mRNA expression levels of candidate genes. The results showed that the inhibition of HepG2 proliferation was more obvious with the increase of HQP concentration. When the concentration of HQP was 100 μg·mL-1, the necrosis rate increased to 18.78%, and the number of red necrotic cells stained with PI was observed under the microscope. The Western blot results showed an increase in RIP1 protein levels. The results of RNA-seq analysis showed that 26 000 related genes were regulated by HQP, and 979 genes were more regulated. KEGG analysis found that some differentially expressed genes were associated with p53 signaling pathway, and qRT-PCR further verified that the sequencing results were reliable. HQP may cause programmed necrosis of HepG2 cells and may be involved in the p53 signaling pathway.
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We studied the effect of aqueous extract from Huang qi on gene expression profile of doxorubicin induced nephropathy in rats, and explored the molecular mechanism of the intervention. The gene expression profiles of control group, model group and aqueous extract from Huang qi group were detected by using transcriptome sequencing technique. The differentially expressed genes (DEGs) were screened by STEM trend analysis software. GO function enrichment and KEGG pathway analysis were performed for DEGs, and the gene expression level was verified by real-time fluorescence quantitative PCR (RT-qPCR). The results showed that, compared with the control group, 432 DEGs were obtained in doxorubicin nephropathy model group; compared with the model group, 811 DEGs were obtained due to aqueous extract of Huang qi. The results of GO function enrichment and KEGG enrichment analysis indicated that PI3K-AKT pathway (Col6a6, Nr4a1, Sgk1, Gng7) and lipid metabolism-related genes (Cpt1b, Pcsk9, Abca1, Ascm5) were the key pathways and genes in the treatment of doxorubicin induced nephropathy by aqueous extract from Huang qi, which played a protective role in kidney. In conclusion, the molecular mechanism of aqueous extract from Huang qi in protection against doxorubicin induced nephropathy rats is closely related to apoptosis-related genes and lipid metabolism-related genes, suggesting for the need of follow-up study for key gene validation and mechanism of action of aqueous extract from Huang qi for prevention of doxorubicin induced nephropathy.
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This study was aimed to observe the effect of Bai-Zhu Huang-Qi (BZHQ) decoction ethyl acetate extract on NOD like receptor family,pyrin domain-containing 3 (NLRP3) inflammasome in macrophages.The U937 cells were pretreated with phorbol 12-myristate 13-acetate (PMA,10 ng· mL-1) for 48 hours to induce macrophages.Effects on cell viability by different doses of BZHQ decoction ethyl acetate extract (0,3.125,6.25,12.5,25,50,100 μg· mL-1) were observed to select the appropriate concentration.Contents of NLRP3 and caspase-1 in cells were detected by real-time PCR and western blot.The concentration of interleukin-1β (IL-1β) in cell supernatant was detected by enzyme linked immunosorbent assay (ELISA).The cell counting kit-8 (CCK-8) assay showed that when the drug concentration was lower than 25 μg· mL-1,there was no impact on cell viability;when the drug concentration was higher than 50 μg· mL-1,there was inhibition on cell viability (P ＜ 0.05).The concentration of 25 μg· mL-1 was used to conduct the following experiment.Compared to the blank group,the expression of NLRP3 and caspase-l in cells of the LPS group were significantly increased (P ＜ 0.01).The concentration of IL-1β in cell supernatant was also significantly increased (P ＜ 0.01).After treated with BZHQ decoction ethyl acetate extract,levels of NLRP3,caspase-1 and IL-1β were significantly decreased (P ＜ 0.05).It was concluded that BZHQ decoction ethyl acetate extract can inhibit the production of NLRP3 inflammasome in LPS-stimulated macrophages.
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Objective To investigate the therapeutic effect of astragalus injection onβcells apoptosis of type 1 diabetes mice. Methods 32 mice were randomly divided into a control group, a diabetes group, a small dose group and a large dose group of astraglus. Except the control group, mice in the other 3 groups received intraperitoneal injection of STZ in order to induce diabetic mellitus. Then one week after injection of STZ, mice in the small dose group and the large dose group of astraglus were given 30, 60 g/(kg?d) doses of astragalus injection, while the other 2 groups were injected into an equal volume saline for 4 weeks. Serum NO, inducible nitric oxide synthase (iNOS) and insulin levels were detected by enzyme linked immunosorbent assay method, and apoptosis was measured by using a TUNEL assay. Results Compared with the diabetes group, the serum NO (25.81 ± 2.09μmol/L, 18.84 ± 3.95μmol/L vs. 30.34 ± 2.53μmol/L) and iNOS (21.38 ± 4.48μmol/L, 17.00 ± 3.05μmol/L vs. 26.62 ± 2.48μmol/L) levels were significantly reduced in the small dose group and the large dose group of astragalus (P0.05). Conclusion Astragalus injection can reduce the iNOS activity and NO production, but can not effectively decreasingβcells apoptosis.
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Objective To dynamically the preparation process of Huangqi injection and to verify the rationality and existing problems of the process. Methods The preparation was made by the current standard (WS3-B-3335-98) issued by Ministry of Health, and the solid amount of the key processes were measured. The HPLC separation was performed on a Agilent Zorbax Bio-C18 reversed-phase column (250 mm × 4.6 mm, 5μm) in gradient mode of acetonitrile-water with UV detection at 254 nm. The column temperature was kept at 25 ℃, and the flow rate of mobile phase was 1.0 ml/min. Results Solid amounts of different operations of Huangqi injection were measured accurately. The content of the third water extracts was only 6.1%, and the changes of HPLC pointed the content of the 12 peaks of the second peaks decreased obviously. Conclusion The technological rationality of Huangqi injection need to be verified and optimized, the dynamic analysis of HPLC describes the changes of chemical constituents of Huangqi injection qualitatively, which also provides a reference value for establishing its fingerprint.
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ObjectiveTo evaluate the therapeutic effect ofHuangqi-Guizhi-Wuwu decoction in patients with diabetic peripheral neuropathy (DPN).MethodsA total of 78 patients with DNP were included and divided into a control group and a treatment group, with 39 in each group. The patients in the control group were administrated with appropriate exercise, control diet, hypoglycemics, mecobalamin and vitamin B1, those in the treatment group were receivedHuangqi-Guizhi-Wuwu decoction on the basis of the control group. The patients in both groups were treated for 8 weeks. The serum activities of superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase were determined by enzymatic-colorimetric method, and the serum malondialdehyde level was measured using colorimetric method. Sensory nerve conduction velocity (SCV) and motor nerve conduction velocity (MCV) were measured using an Electromyograph and Evoked Potential Equipment.ResultsAfter treatment, MCV (43.12 ± 4.1 m/svs. 40.27 ± 2.8 m/s;t=3.585,P<0.05)、SCV (41.51 ± 3.4 m/svs. 39.63 ± 3.5 m/s;t=2.406,P<0.05) in the treatment group were significantly increased than those in the control group. The serum activities of SOD (51.1 ± 6.6 U/mlvs. 47.6 ± 3.9 U/ml;t=2.851,P<0.01),GSH-Px (241.7 ± 36.1 U/mlvs. 213.9 ± 31.7 U/ml;t=3.614,P<0.01) in the treatment group were significantly increased than those in the control group. The total efficiency in the treatment group was significantly higher than that in the control group (87.2%vs. 69.2%;χ2=4.333,P=0.033).ConclusionsHuangqi-Guizhi-Wuwu decoction combined with conventional therapy could improve antioxidant capacity and control DPN progress in patients with DPN.
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ObjectiveTo observe the clinical efficacy ofLing Gui Ba Fa(eight methods of the sacred tortoise) plusHuang Qi Jian Zhongdecoction in treating stomachache due to deficiency-cold in spleen and stomach.MethodTotally 100 eligible subjects were randomized into an observation group and a control group. The control group was intervened byHuang Qi Jian Zhong decoction, while the observation group was additionally given acupuncture withLing Gui Ba Faneedling method.ResultTwo weeks later, the symptom and sign score in the observation group was significantly lower than that in the control group (P0.05); the recovery and markedly-effective rate was 88.0% in the observation group versus 62.0% in the control group, and the difference was statistically significant (P<0.01).ConclusionLing Gui Ba Faneedling method can effectively enhance the efficacy ofHuang Qi Jian Zhongdecoction in treating stomachache due to deficiency-cold in spleen and stomach.
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This study was aimed to explore the effect of total flavonoids and saponins from Huang-Qi Ge-Gen (HQGG) decoction on blood glucose (BG), serum lipid, interleukin-12 (IL-12) and interleukin-15 (IL-15) of liver in diabetes mellitus (DM) rats, in order to investigate their interactions in regulating DM processes. A total of 66 SD rats were randomly divided into 6 groups, which were the normal group, model group (A1B1), control group, total flavonoids group (A2B1), total saponins group (A1B2), and total flavonoids and saponins group (A2B2), with 11 rats in each group. Except the normal group, other groups were intraperitoneal injected with streptozotocin (STZ). And the experiment was according to 2×2 factorial design experiment scheme. The BG was determined before STZ injection and 7 days after the STZ injection. After 30 days, BG, serum lipid, IL-12 and IL-15 of liver were tested. Related indexes were calculated to the weighted composite score. Main and interactive effect of total flavonoids and saponins were studied according to the factorial design experiment scheme. The results showed that compared with the normal group, all indexes of model group showed statistical differences (P<0.05). Total flavonoids and saponins from HQGG decoction can effectively reduce BG, without any interactions between them. Both the total flavonoids and total saponins can reduce serum cholesterol (CHO), triglyceride (TG), liver IL-12 and IL-15. And there were interactive effects. The single use of herb achieved better effects than the combination. It was concluded that total flavonoids and saponins from HQGG decoction can reduce BG, CHO, TG, and liver IL-12 and IL-15 levels in rats. However, the regulation of total flavonoids and saponins on indexes mentioned above showed no additive effect.
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Objective To investigate the role of Huang-qi in balance of TH1/TH2 in asthma on dendritic cells level. Methods DCs from human peripheral blood mononuclear cells(PBMC) were induced by rhGM-CSF and rhIL-4, and then were identified. The level of IL-12 and IL-10 produced by DCs were de-tected by ELISA assay. After autoreactive T cells, mRNA of T-bet and GATA-3 was measured by RT-PCR. Simultaneously, IL-4 and IFN-γ were determined by flow cytometer. Results After 7 days culture, IL-12 was significantly decreased in asthma group compared to control group (P ＜ 0.01), whereas IL-10 on the opposite. At the 7th day of co-culture with T cells derived from floating cells, the IFN-γ/and T-bet mRNA level in asthma group were significantly decreased than that in control group, whereas IL-4, GATA-3 mRNA level, the ratio of IL-4/IFN-γ and GATA-3/T-bet were apparently increased in asthma group than that in control group(P＜0.01). After Huang-qi treatment, the IFN-γ/and T-bet mRNA level were significantly in-creased, whereas the ratio of IL-4/IFN-γ and GATA-3/T-bet, and the IL-10 level were apparently de-creased, but the level of IL-12, IL-4 and GATA-3 mRNA were not changed significantly. Conclusion DCs in asthma regulated the balance of TH1/TH2 by means of secreting decreased IL-12 and increased IL-10, that made TH2 playing a dominance role which is the key factor in initiating asthma. Huang-qi regulated DCs through decreasing the level of IL-10, and thus decreased the ability of inhibiting the differentiation of TH1 from TH0, that is also inhibiting the differentiation of TH2 from TH0 directly.
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Objective: To investigate effects of Fang Ji Huang Qi Capsules on the level of blood uric acid and blood pressure in the patient of primary hypertension of type of stagnation of phlegm, Methods: 61 patients were divided into the treatment group (34 cases) treated with Fang Ji Huang Qi Capsules plus Norvasc and the control group (27 cases) treated with Norvasc. They were treated for 6 weeks and blood uric acid level, dynamic blood pressure and TCM clinical symptoms were observed. Results: The blood uric acid level decreased significantly, the valley to peak ratio of dynamic blood pressure increased and cumulative score of TCM clinical symptoms decreased significantly in the treatment group as compared with the control group (P