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Objective To observe the amplification effect of mitochondrial DNA (mtNDA) on the inflammatory response of rat alveolar macrophage induced by lipopolysaccharide (LPS), and to preliminarily explore its mechanism. Methods mtDNA of Sprague-Dawley (SD) rat liver tissue was harvested, ultra-micro spectrophotometer and 1% agarose gel electrophoresis were used to detect the concentration and quality of mtDNA. The alveolar macrophages of SD rat were isolated and cultured, the macrophages in logarithmic growth phase were divided into phosphatic buffer solution (PBS) group, LPS group, mtDNA group and LPS+mtDNA group. The first three groups were added equal volumes of PBS, LPS 1 mg/L or mtDNA 10 mg/L to the alveolar macrophages medium for 6 hours and 12 hours, respectively; the alveolar macrophage medium of LPS+mtDNA group was stimulated with 1 mg/L LPS for 6 hours and then stimulated with 10 mg/L mtDNA for 6 hours. The levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the cell supernatant were detected by enzyme linked immunosorbent assay (ELISA);the expression of the key protein of Pyroptosis-Gasdermin D (GSDMD) was detected by Western Blot. Results ① The mtDNA A260/280 ratios were between 1.8-2.0, and agarose gel electrophoresis showed a single band, with a size of about 16 kb. ② After alveolar macrophages stimulated by LPS or mtDNA for 6 hours or 12 hours, respectively, the levels of IL-1β and TNF-α were higher than those in PBS group. When cells were treated with mtDNA for 6 hours after LPS stimulation 6 hours, the levels of IL-1β were higher than those in LPS 12 hours group (ng/L: 366.27±23.35 vs. 154.70±23.32, 1 < 0.01), but the levels of TNF-α had no significant difference compared with LPS 12 hours group (ng/L: 836.13±25.01 vs. 802.67±30.48, 1 > 0.05). ③ The protein expressions of GSDMD in LPS group, mtDNA group and LPS+mtDNA group were significantly higher than those in PBS group (GSDMD/β-actin: 1.77±0.05, 1.65±0.04,2.40±0.05, 1.00±0.02, all 1 < 0.01), and the protein expression of GSDMD in LPS+mtDNA group was higher than that in LPS group (1 < 0.01). Conclusion mtDNA amplifies LPS-induced alveolar macrophage inflammatory responses,which mechanism may be related to the increase in pyroptosis mediated by mtDNA.
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Objective To investigate the effect of Terpinen-4-ol on the proliferation of lung adenocarcinoma A-549 cells and its related mechanism. Methods A-549 cells were treated with different concentrations of Terpinen-4-ol. The inhibitory effect of Terpinen-4-ol on A-549 cells was tested by MTT method. Cell grow ability was determined by CCK-8 colorimetry. The ultrastructure of A549 cells were observed by transmission electron microscopy before and after Terpinen-4-ol treatment. The changes of cell cycle, apoptosis, and the level of intracel-lular calcium were inspected by flow cytometry. Inoculated the lung adenocarcinoma A-549 cells on the nude mice to form transplantation tumor. The experimental nude mice with transplantation tumors were divided into three groups:negative control group,high dose positive con-trol group and low dose positive control group. The mice were given continuously intraperitoneal injection for 10 days, and then the transplan-tation tumors were taken and the size and weight of them were detected. Results After Terpinen-4-ol treatment for 24 h,MTT assay showed that the IC50 value of A549 cells was 0. 067% v/v. The growth curves of positive control groups were significantly smooth than the negative control group. The formation of autophagosome increased after treatment with Terpinen-4-ol. The results of flow cytometry showed that the cell cycle was arrested in S phase,Terpinen-4-ol could induce apoptosis of A549 cell, The intracellular calcium concentrations in positive control groups were significantly higher than the negative control group(P<0. 05). Low dose group and high dose group restrained the growth of the transplantation tumor obviously, and the tumor inhibitory rate were 53. 33% and 77. 76% respectively. Conclusion Terpinen-4-ol has inhibitory effect on the proliferation of A-549 cells in vitro and in vivo.
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Objective To prepare TF conjugated doxorubicin and Rg 3 loaded liposome and evaluate their properties and effect on the treatment of gastric cancer. Method The liposomes were prepared by thin iflm hydration method. The efifciency of cellular uptake on MKN-28 cells in vitro was evaluated. The anti-proliferation efifciency of TF-LP-DOX/Rg 3 was evaluated by MTT assay. Tumor spheroids were used to evaluate anti-tumor ability of TF-LP-DOX/Rg 3. Results The result demonstrated that TF-LP uptaken by MKN-28 were 2.9 times higher than that of LP(P<0.01). The MTT assay and the inhibition of tumor spheroids in vitro conifrmed strong inhibitory effect of TF-LP-DOX/Rg 3. Conclusion TF-LP-DOX/Rg 3 easily prepared and it is a potential delivery system for the treatment of gastric cancer.
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This review addresses the latest situations and advances of near infrared spectroscopy (NIRS) in which detection of counterfeits and imitations, as well as monitoring origin and quality of Chinese crude drugs and Chinese patent medicines (CCDM) through consultation and summarization of relative literatures. On the one hand, NIRS gradually reveals its advantages and discriminating ability in the ways of nondestructive, rapid, simple, easy, and handy assessment. However NIRS still has some problems in representative samples and models stability for practice of CCDM. In order to keep up with popularization of NIRS in other areas, applications in detection of precious and/or priceless herbals, on-line quality control of valuable herbs, and screening of some chemicals illegally mixed into herbal preparations may be focused preferentially.
Subject(s)
Drugs, Chinese Herbal , Chemistry , Medicine, Chinese Traditional , Spectroscopy, Near-Infrared , MethodsABSTRACT
While tracing back the 50 years of history and achievements in higher education of TCM,it compares it with western medicine,and sets up 3 elements(research object,knowledge system and intellectual producing style)scale for a separate discipline.Chinese medicine development is slow and weak;therefore it expounds the self-enhancement path from adjusting key Chinese medicine development points,strengthening the idea of paddling its own canoe,perfecting knowledge system and improving the academic and scholar level of Chinese medicine team.
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The geo-herbalism of Cangzhu, Atractylodes lancea (Thunb.) DC. , was studied for itsstrict quality control. Samples of typical and representative populations of geo-herbal and non-geo herbalorigin were collected, examined and cultivated through field work. Volatile oils from samples of varioushabitats were determined by GC/MS, and their respective bacteriostatic minimum inhibitory concentration(MIC)determined. Results of the study showed that both the size of capitulum and cleft on the leaves var-ied greatly even in species of the same population. The main components in their volatile oil also variedgreatly with Maocangzhu (MCZ) contains atractylone (29.54%) and atractylodin (17.82%); that of Nan-cangzhu (NCZ) contains hinesol (61.86%) and β-eudesmol (19.79%), and Baicangzhu (BCZ) cantains βeudesmol or mixture of β-eudesmol and atractylone (over 30%). Volatile oils of speciaes from Luotina,Taibai, and Huanglong showed potent bacteriostatic activity against Streptococcus hemolyticus (SH), com-parable to penicillin or streptomycin. In conclusion, A. lancea is a polymorphic complex greatly influencedby geographic environment and a reasonable proposal is to name them by geo-herbal names such as: MCZas A. lancea var. maoshanensis Hu et Feng; NCZ as A. lancea subsp. luotianensis Hu et Feng; and BCZ asA. lancea var. chinensis (Bunge) Kitam other than independent species as A. chinensis Bunge. It was in-correct to say that MCZ has no atractylone in its volatile oil and therefore differs from A. macrocephalaKoidz. (Baizhu). Inhibitory activity of the volatile oils against SH and Pseudomonas aeruginosa (PA) arealso quite different due to the habitats and imply that the rhizome from Luotian may be used to cure infec-tion diseases caused by SH and PA besides their traditional uses.
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Object To develop a new method of discriminating the principle components of Radix Paeoniae Rubra (RPR) from different areas, and to find the reason for forming high-quality of RPR in the place of the genuine. Methods Using Foruier transform infrared (FTIR) absorption spectrometry, the characteristic peaks of fingerprint infrared spectra of RPR samples from 18 habitats were recognized and compared. Results Frequency, intensity and shape of infrared absorption spectra were obviously different between wild and cultivated groups of RPR. The infrared absorption peaks of RPR in Duolun, a famous Chinese orthodox drug, were of distinct characteristics. Conclusion FTIR technique is first applied to rapid analysis of principle component of RPR from different areas. So an operable method in the quality control and discrimination of RPR in the place of the genuine is provided.