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主 办:北 京 中 医 药 大 学
ISSN 1006-2157 CN 11-3574/R

Table of Content

    30 November 2018, Volume 41 Issue 11 Previous Issue    Next Issue
    On inflammation-cancer transformation of chronic gastritis from view of traditional Chinese medicine*
    Ding Xia, Wei Wei, Shen Hong, Sun Zeqi
    2018, 41 (11):  885-889.  doi: 10.3969/j.issn.1006-2157.2018.11.001
    Abstract ( 551 )   PDF (1132KB) ( 426 )   Save
    The inflammation-cancer transformation of chronic gastritis is a classical model of malignant transformation from chronic uncontrollable inflammation cancer, and traditional Chinese medicine (TCM) plays an important role in delaying, blocking and reversing the progress of this disease. From the perspective of TCM, TCM cognition on inflammation-cancer transformation of chronic gastritis, and the function, value, bottleneck problem and countermeasure of TCM in controlling thus progress are expounded. At present stage, there are relatively few TCM literatures on discussion of inflammation-cancer transformation of chronic gastritis, and no systematic research Results achieved. The TCM theory of “dynamic thinking” and “preventive treatment of disease” plays an important role in the prevention and treatment of inflammation-cancer transformation of chronic gastritis. TCM has advantages in removing causes of disease, alleviating patients’ clinical symptoms and improving the pathological morphology of gastric mucosa. In the process of controlling inflammation-cancer transformation of chronic gastritis, TCM has the characteristics of higher safety, lower drug dependence and good compliance. It is of great significance and value to control the progress of the disease and reduce the incidence of gastric cancer. However, there are still some problems including a lack of systematic ideas of differentiation and treatment in clinical practice, unclear time for intervention of inflammation-cancer transformation of chronic gastritis, and indistinct combined effects with Western medicine at the present stage. The bottleneck problems are manifested as program in scientific research cannot reflect TCM characteristics, systematic mechanism exploration is in shortage, and research Results are difficult to be applied in clinical practice. Therefore, further consideration and measures are needed in controlling inflammation-cancer transformation of chronic gastritis by TCM
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    Inheritance and development of spleen-stomach theory in Huangdi Neijing (Yellow Emperor’s Internal Classic) by Huang Yuanyu*
    Gao Zhili, He Juan
    2018, 41 (11):  890-893.  doi: 10.3969/j.issn.1006-2157.2018.11.002
    Abstract ( 562 )   PDF (1052KB) ( 1385 )   Save
    Spleen-stomach theory is an important academic thought in Huangdi Neijing (Yellow Emperor’s Internal Classic). The idea of emphasizing earth was a concrete embodiment of Chinese philosophy in traditional Chinese medicine at that time. Later generations of doctors inherited and developed spleen-stomach theory in Huangdi Neijing, among them, Huang Yuanyu, a physician in the Qing Dynasty, was a particular representative. Huang’s theory of qi circumfluence and four images of earth pivot emphasizes the up-downs and rotation of qi in the middle earth, and stresses ascending of the left-side qi movement. He proposed that damp earth, cold water and wood depression were main pathogenesis of human diseases, which has important influence on later clinical practice.
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    Brief account of Zhang Zhongjing’s medical origin*
    Pan Zhongyi, Fu Yanling, Song Jia, Ni Shenglou
    2018, 41 (11):  894-899.  doi: 10.3969/j.issn.1006-2157.2018.11.003
    Abstract ( 497 )   PDF (1123KB) ( 808 )   Save
    Zhang Zhongjing medicine has an extraordinary position and influence in the academic development history of traditional Chinese medicine (TCM). Those who have studied TCM since ancient times have absorbed nutrient from it. To study Zhongjing medicine, we should first understand its origin and development. In this paper, we comb up this issue from the following three aspects including the origin of Zhongjing medicine, the emergence of Zhongjing medicine, and the branch of Zhongjing medicine. We believe that the origin of Zhongjing medicine can be dated back to Sanshi Medicine in ancient times and Four Medical Schools in the Spring and Autumn Warring States period, the emergence of Zhongjing medicine is reflected as integration of medical classics and classical formulas and coherence of every section of TCM pattern differentiation and treatment, and the branch of Zhongjing medicine is mainly manifested as continuous development of Shanghan schools during 1 800 years, its important influence on other medical branches and schools, as well as the spread and changes abroad.
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    Characteristic dose-effect relationship of Chinese medicinals*
    Wu Tong, Jia Chunhua
    2018, 41 (11):  900-904.  doi: 10.3969/j.issn.1006-2157.2018.11.004
    Abstract ( 399 )   PDF (1058KB) ( 195 )   Save
    Characteristic dose-effect relationship of Chinese medicinal studied in this paper refers to the relationship between dose and effect, that is, changes in efficacy caused by changes in drug dosage. By collecting ancient medical literature based on Chinese herbal books in all ages in Chinese Medical Dictionary, and searching CNKI and WanFang Data, the Chinese medicinals with such dose-effect relationship are collected, summarized and analyzed. The understanding of the characteristic dose-effect relationship of Chinese medicinals has gone through 3 stages of development, that is, such dose-effect relationship was directly applied in clinic before the Song and Yuan Dynasties, and was clearly expressed in the Ming Dynasty and further enriched in the Qing Dynasty. After the introduction of Western medicine into China, the dose-effect relationship of some Chinese medicinals has undergone verification by modern physicians through experiments or mechanism analysis through medicinal chemistry, which improves the development of research on dose-effect relationship of Chinese medicinal.
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    Intervention to PI3K/AKT signaling pathway by Buqi Huoxue Tongluo Fang in rats with idiopathic pulmonary fibrosis*
    Zhang Yanxia, Shi Liqing, Yang Xiaohua, Shi Lei
    2018, 41 (11):  905-909.  doi: 10.3969/j.issn.1006-2157.2018.11.005
    Abstract ( 347 )   PDF (2634KB) ( 175 )   Save
    Objective To investigate the influence of Buqi Huoxue Tongluo Fang (Qi-tonifying Blood-activating Collateral-freeing Formula, BHTF) on PI3K/AKT signaling pathway in rat model of idiopathic pulmonary fibrosis (IPF). Methods All rats were randomly divided into 6 groups including normal group, model group, prednison group, low-dose, mid-lose and high-dose BHTF groups. Except of normal group, other groups were given bleomycin (3 mg/kg) for modeling. After 2 d, all BHTF groups were orally given BHTF decoction in different doses (8.25 g/kg, 16.5 g/kg and 33 g/kg), prednison group was orally given prednisone acetate suspension (3.6 mg/kg), and normal group and model group were orally given equivalent normal saline. The changes of rat general status were observed, difference in protein expressions of PTEN, PI3K, AKT and mTOR signaling pathway were detected by using immunohistochemistry technique, and difference in genetic expressions was observed by using RT-PCR at 2 time points (14 d and 28 d). Results Compared with model group and prednison group, the rat general status and weight were improved (P<0.05), PTEN expression was up-regulated and expressions of PI3K, AKT and mTOR were down-regulated in all BHTF groups (P<0.05). Conclusion BHTF can play an anti-fibrosis role through intervening the activation of PI3K/AKT signaling pathway.
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    Influence of Qingre Liangxue Fang on imbalance of Th17/Treg cells in psoriasiform mouse model induced by imiquimod and interventional effect of miR-210 agonist*
    Li Nuo, Ya Lijing, Wang Lei, Wang Ying, Bai Yanping
    2018, 41 (11):  910-917.  doi: 10.3969/j.issn.1006-2157.2018.11.006
    Abstract ( 347 )   PDF (2807KB) ( 215 )   Save
    Objective To observe the influence of Qingre Liangxue Fang (Heat-clearing Blood-cooling Formula, QRLXF) on imbalance of Th17/Treg cells in psoriasiform mouse model induced by imiquimod and interventional effect of miR-210 agonist (AgomiR-210). Methods Female BALB/c mice (n=42) were randomly divided into 7 groups: normal group, model group, high-dose, mid-dose and low-dose QRLXF groups (30.2 mg /kg, 15.1 mg /kg, 7.05 mg /kg), and high-dose and low-dose AgomiR-210 groups (8 nmol/L, 4 nmol/L). The dynamic changes of skin lesions were observed by using scores of psoriasis area and severity index (PASI) in psoriasiform mouse model. The pathological changes in skin lesion tissue were observed by using light microscope. The expressions of Foxp3 and RORγt were detected by using immunohistochemistry technique, and expression of miR-210 was detected by using qRT-PCR. Results The skin lesions, pathological changes and PASI scores were significantly improved by QRLXF showing dose-effect dependence, and the improvement of PASI scores was the most significant in high-dose QRLXF group compared with model group (P<0.001). The pathological Results showed that AgomiR-210 reduced the therapeutic effect of QRLXF, which was more significant in low-dose QRLXF group compared with high-dose QRLXF group (P<0.01). The Results of immunohistochemistry technique showed that QRLXF up-regulated Foxp3 expression and down-regulated RORγt expression, and AgomiR-210 down-regulated Foxp3 expression and up-regulated RORγt expression. The Results of qRT-PCR showed that QRLXF inhibited miR-210 mRNA expression compared with model group (P<0.01). AgomiR-210 up-regulated miR-210 mRNA expression compared with high-dose QRLXF group (P<0.01). Conclusion QRLXF can significantly relieve the skin lesions in psoriasiform mouse model possibly through inhibiting miR-210 expression, up-regulating Foxp3 expression, down-regulating RORγt expression, and ameliorating imbalance of Th17/Treg cells.
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    Improvement of cognitive ability by Jianpi Huatan Huoxue Fang and possible mechanism in rats with subclinical hypothyroidism*
    Chen Wei, Yang Xiao, Zhang Fengnuan, Shun Qunqun, Cha Yi, Gao Tianshu
    2018, 41 (11):  918-927.  doi: 10.3969/j.issn.1006-2157.2018.11.007
    Abstract ( 363 )   PDF (3076KB) ( 235 )   Save
    Objective To study the influence of Jianpi Huatan Huoxue Fang (Spleen-fortifying Phlegm-resolving Blood-activating Formula, JHHF) on cognitive ability in rats with subclinical hypothyroidism (SCH), and discuss possible mechanism. Methods SCH rat model was established by total thyriodectomy and postoperative hypodermic injection of L-T4 in male Wistar adult rats. The rats were randomly divided into model group, low-dose group (13.7 g/kg of JHHF), high-dose group (41.1 g/kg of JHHF), L-T4 group (5.0 μɡ/kg), low-dose+L-T4 group (13.7 g/kg/d of JHHF +5.0 μɡ/kg/d), and high-dose+L-T4 group (41.1 g/kg/d of JHHF+5.0 μɡ/kg/d), and a sham-operation group was established at the same time. The treatment duration was for 6 weeks. After 6 weeks, all groups were given Morris water maze (MWM) test, and rats were executed. The levels of serum T4 (TT4) and thyroid stimulating hormone (TSH) were detected by using respectively radioimmunoassay (RIA) and immunochemiluminometric assay (ICMA). The Nissl bodies were observed in hippocampal CA1 zone by using light microscope after Nissl staining. The level of brain-derived neurotrophic factor (BDNF) in hippocampus was detected by using ELISA. The expressions of BDNF and its receptor-TrKB were detected by using RT-qPTC, and BDNF+TrKB were detected by using double-labelling immunofluorescence. Results The level of serum TSH increased significantly (P<0.01), TT4 level had no significant changes (P>0.05), the time of locating cruise and times of space exploration decreased significantly (P<0.01), and hippocampal BDNF level and mRNA expressions of BDNF and TrKB decreased significantly (P<0.01) in model group compared with sham-operation group. The level of serum TSH decreased significantly (P<0.05), the time of locating cruise and times of space exploration increased significantly (all P<0.05), hippocampal BDNF level and mRNA expressions of BDNF and TrKB increased significantly (all P<0.05), and numbers of double positive cells of BDNF/TrKB and hippocampal Nissl bodies increased significantly (all P<0.05) in all JHHF groups compared with model group. The curative effect of JHHF was better than that of L-T4, and curative effect of JHHF combined with L-T4 was the best. Conclusion JHHF can improve learning and cognitive abilities in SCH rats, and the mechanism may be related to the regulation of thyroid function and hippocampal BDNF. The combination of JHHF and L-T4 has synergistic effect.
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    Influence of Qingqianliuye (leaf of Cyclocarya paliurus) on skeletal muscle inflammatory response, insulin pathway proteins and glycogen synthesis in T2DM rats*
    Yao Junkai, Gao Xuemin, Zhang Jianjun, Li Wei, Fu Lu, Wang Leilei, Jia Lan, Zhao Xiuting, Wang Jingxia
    2018, 41 (11):  928-934.  doi: 10.3969/j.issn.1006-2157.2018.11.008
    Abstract ( 336 )   PDF (1097KB) ( 177 )   Save
    Objective To study the influence of Qingqianliuye (leaf of Cyclocarya paliurus) aqueous extract (CPAE) on skeletal muscle inflammatory response, insulin pathway proteins and glycogen synthesis in rats with type 2 diabetes mellitus (T2DM). Methods T2DM rat model was established by high-fat diet for 8 weeks and intraperitoneal injection of low-dose STZ (30 mg/kg). The control rats were included into normal group, and modeled rats were divided into model group, metformin group, low-dose, mid-dose and high-dose CPAE groups. The normal group and model group were given 0.9% NaCI solution, metformin group was given metformin suspension (0.3 g/kg), and CPAE groups were given, respectively, CPAE (0.25 g/kg, 0.5 g/kg and 1 g/kg) for 4 weeks. After executed rats, the levels of free fatty acid (FFA) and glycogen in skeletal muscle homogenate were detected by using chromatoptometry. The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and interleukin-1β (IL-1β) were detected by using radioimmunoassay (RIA). The content of IκB kinase (IKK), nuclear factor-κB (NF-κB), c-Jun N-terminal kinase 1 (c-JNK1), insulin receptor substrate 1 (IRS1), phosphorylated insulin receptor substrate 1 (P-IRS1), phosphatidylinositol 3-kinase (PI3K), glucose transporter 4 (GLUT4) and glycogen synthase (GS) were detected by using ELISA. Results Compared with model group, the levels of FFA, TNF-α, IL-6 and IL-1β decreased significantly in all CPAE groups (P<0.05, P<0.01). The content of IKK, NF-κB and JNK1 proteins of inflammatory pathway decreased significantly in all CPAE groups (P<0.05, P<0.01). The protein expressions of IRS1, P-IRS1, PI3K, GLUT4 and GS, and glycogen content increased significantly in all CPAE groups (P<0.05, P<0.01). Conclusion CPAE raises the content of skeletal muscle insulin pathway proteins and glycogen synthesis protein, inhibits expressions of inflammatory pathway proteins, and improves glycogen synthesis in T2DM rats. The mechanism may be related to that CPAE can reduce content of skeletal muscle pro-inflammatory factors, inhibit expressions of inflammatory pathway proteins, and relieve metabolic inflammatory response.
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    Influence of genistein in high and low concentration on growth and ERα/ERβ protein ratio of Ishikawa cells*
    Cheng Ran, Liu Xiaoli, Xue Xiao’ou, Xie Wei
    2018, 41 (11):  935-942.  doi: 10.3969/j.issn.1006-2157.2018.11.009
    Abstract ( 326 )   PDF (1261KB) ( 234 )   Save
    Objective To study the difference of genistein between high and low concentration on the proliferation, colony formation and protein expression ratio of ERα to ERβ (ERα/ERβ) in Ishikawa cells of human endometrial carcinoma with positive estrogen receptor (ER). Methods ER-positive Ishikawa cells of human endometrial carcinoma were taken as medium research objects. Before formal experiments, the cells were cultured in Opti-MEM phenol-free serummedium for 24 h, and then divided into low-dose genistein group, high-dose genistein group and solvent control group. The genistein solution and solvent control were prepared by the Opti-MEM phenol-free serum medium containing 2% DCS-FBS and were sequentially added to 3 groups. The influence of high-concentration genistein (10 μmoL/L, 25 μmoL/L, 50 μmoL/L) and low-concentration genistein (0.001 μmoL/L, 0.01 μmoL/L, 0.1 μmoL/L, 1 μmoL/L) on proliferation of Ishikawa cells was detected by using cell counting kit-8 (CCK-8) after intervening for 24 h, 48 h and 72 h. The influence of high-concentration genistein (25 μmoL/L, 50 μmoL/L) and low-concentration genistein (0.01 μmoL/L, 0.1 μmoL/L) on colony formation was detected by using colony formation assay after intervening for 1 week. The influence of high-concentration genistein (25 μmoL/L, 50 μmoL/L) and low-concentration genistein (0.01 μmoL/L, 0.1 μmoL/L) on protein expression ratio of ERα/ERβ by using Western blotting assay after intervening for 48 h. Results Low-concentration genistein (0.001 μmoL/L, 0.01 μmoL/L, 0.1 μmoL/L) promoted the proliferation of Ishikawa cells, which was the most significant after intervening for 48 h, and proliferation promoting effect was the highest when genistein concentration reached 0.1 μmoL/L (P<0.01). High-concentration genistein 10 μmoL/L, 25 μmoL/L, 50 μmoL/L inhibited the proliferation of Ishikawa cells and showed time-concentration dependence. The inhibitory effect reached the highest when genistein concentration was 50 μmoL/L after 72 h. The colony formation rate was 14.60% and 15.13% respectively in low-concentration genistein groups compared with solvent control group (12.20%, P<0.05). The colony formation rate was 6.07% and 4.27% respectively in high-concentration genistein groups, which was significantly lower than that in solvent control group (P<0.01). The protein expression of ERα was up-regulated in high-concentration genistein groups and low-concentration genistein groups, and the expression reached the highest when genistein concentration was 0.1 μmoL/L. The protein expression of ERβ was down-regulated, and the expression reached the lowest when genistein concentration was 25 μmoL/L or 50 μmoL/L. The protein expression ratio of ERα/ERβ was significantly up-regulated, and reached the highest when genistein concentration was 0.1 μmoL/L. Conclusion Low-concentration genistein has effect of promoting Ishikawa cells with positive ER, which may be related to up-regulation of protein expression ratio of ERα/ERβ. High-concentration genistein can also up-regulate protein expression ratio of ERα/ERβ, but may play the role of inhibiting growth of Ishikawa cells through non-ER dependent mechanism.
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    Protective effect of Honghua (Safflower, Flos Carthami) on acute liver injury induced by carbon tetrachloride in rats and mechanism study*
    Lyu Xiaomei, Lu Renling, Ma Yuehong, Ma Lijie
    2018, 41 (11):  943-949.  doi: 10.3969/j.issn.1006-2157.2018.11.010
    Abstract ( 346 )   PDF (1519KB) ( 223 )   Save
    Objective To study the protective effect of Honghua (Safflower, Flos Carthami) on acute liver injury induced by carbon tetrachloride (CCl4) in rats and discuss the mechanism. Methods Male Wistar rats (n=50) were randomly divided into normal group, model group, and high-dose, mid-dose and low-dose Honghua groups (each n=10). The Honghua groups were orally given Honghua solution respectively in dose of 1 g/kg, 0.5 g/kg and 0.25 g/kg for 8 d. Except of normal group, other groups were given once intraperitoneal injection of CCl4 for inducing acute liver injury 1 h after the last medication. After 24 h, the rats were executed to collect blood samples for detecting the vitalities of serum ALT, AST and ALP. The liver tissue samples were collected to detect the vitalities of MDA, CAT, GSH-Px and T-AOC after HE staining. The levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), protein expressions JNK, p-JNK and p-c-Jun in JNK signaling pathway, and Caspase-3 and cleaved Caspase-3 were detected by using Western blotting assay. The mRNA and protein expressions of death receptor pathway apoptosis-related factors, FasL and Fas, and mitochondrial apoptosis-related factors: Bax and Bcl-2, were detected by using respectively RT-PCR and Western blotting assay. Results After acute liver injury, the levels of serum ALT, AST and ALP and liver MDA increased significantly (P<0.05), and levels of CAT, GSH-Px and T-AOC decreased significantly (P<0.05) in model group compared with normal group, and all these indexes had statistical difference in all Honghua groups compared with model group (P<0.05). HE staining showed that there were majority of fusion necrosis around the central vein and lots of apoptotic bodies in model group, and apoptotic bodies and small focal necrosis decreased in all Honghua groups. The protein expressions of p-JNK, p-c-Jun and cleaved Caspase-3 increased significantly (P<0.05), mRNA and protein expressions of Bax, FasL and Fas increased significantly (P<0.05), and mRNA and protein expressions of Bcl-2 decreased significantly (P<0.05) in model group compared with normal group. The protein expressions of all inflammatory factors had statistical difference between all Honghua groups and model group (P<0.05), and protein expressions of Bax, p-JNK, p-c-Jun, FasL, Fas and Bcl-2 in JNK signaling pathway had statistical difference between all Honghua groups and model group (P<0.05). The mRNA expressions of Bax and Bcl-2 had statistical difference in mid-dose and low-dose Honghua groups compared with model group (P<0.05), and there was no statistical difference between high-dose Honghua group andmodel group (P>0.05). Conclusion Honghua has a protective effect on acute liver injury induced by CCl4 in rats possibly through antioxidant, anti-activation of inflammatory factors, inhibiting activation of JNK signaling pathway and decreasing apoptosis induced by activation of JNK signaling pathway.
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    Influence of osthole on RAW264.7 cells differentiating to osteoclasts and mechanism study*
    Wang Lining, Ma Yong, Zheng Suyang, Pan Yalan,Tu Pengcheng, Sun Jie, Guo Yang
    2018, 41 (11):  950-958.  doi: 10.3969/j.issn.1006-2157.2018.11.011
    Abstract ( 342 )   PDF (1623KB) ( 166 )   Save
    Objective To investigate the influence of osthole (OST) on RAW264.7 cell lines of mouse monocytes differentiating to osteoclasts and study the related mechanism. Methods Osteoclasts were induced and cultured in vitro by using RAW264.7 cell lines, and then RAW264.7 cell line was divided into negative group, control group and OST group. RAW264.7 cell lines were induced to differentiation by using RANKL, and OST group was intervened with high-dose, mid-dose and low-dose OST (100 μmol/L, 10μmol/L, 1 μmol/L, high-dose, mid-dose and low-dose OST groups) for corresponding time. CCK-8 method was used to screen non-cytotoxic medicinal concentration group, and anti-tartaric acid phosphatase (TRAP) staining method was used to determine number of TRAP positive cells and fusion index. The influence of OST on bone absorptive function was reviewed through detecting area of dissolving pit and fluorescence intensity in bone absorption board. The influence of OST on expressions of nuclear factor of activated T cell (NFAT) and nuclear factor-κB (NF-κB) in RAW264.7 cell lines were observed by using luciferase reporter gene method. The influence of OST on expressions of nuclear factor of activated T cell-1 (NFATc1), CTSK, MMP-9, TRAP, INTE-BETAβ3 and c-Src was detected by using q-PCR. The influence of OST on expressions of p65, IκB, p-IκB and p65 in NF-κB signaling pathway was detected by using Western blotting assay. Results There were 2 medicinal concentration groups (1 μmol/L and 10 μmol/L, low-dose group and mid-dose group) screened by using CCK-8 method. TRAP positive cells and fusion index were significantly lower in all OST groups than those in control group (P<0.05), which was more significant in mid-dose OST group (P<0.05). The related area of bone absorption and fluorescence intensity were significantly inhibited in OST group (P<0.05), which was more significant in mid-dose OST group (P<0.05). OST inhibited significantly the initiations of NFAT and NF-κB (P<0.05) and expressions of osteoclast related genes including NFATc1, and except of Integrin-BETA3 and c-Src, other related genes had statistical difference between low-dose group and mid-dose group (P<0.05). Compared with control group, the protein expression of p-IκB was inhibited significantly (P<0.05), expressions of IκB and p65 were improved (P<0.05), and protein expression of p65 was inhibited (P<0.05) in all OST groups. Conclusion OST can induce down-regulations of NFATc1 and related transcription factors to control differentiation of osteoclasts through inhibiting expression of NF-κB signaling pathway.
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    Anti-inflammatory effect of n-butanol extracts from couplet medicinals of Jingjie and Fangfeng: a study on NF-κB signal pathway mechanism*
    Liu Xiaobo, Sang Wentao, Shi Boyu, Rao Zhili, Luo Jie, Lyu Hongjun, Yang Qingxin, Zeng Nan
    2018, 41 (11):  959-968.  doi: 10.3969/j.issn.1006-2157.2018.11.012
    Abstract ( 372 )   PDF (1997KB) ( 209 )   Save
    Objective To observe the protective effect of n-butanol extracts (NBE) from couplet medicinals of Jingjie (Schizonepeta, Herba Schizonepetae) and Fangfeng (Ledebouriella Root, Radix Ledebouriellae, JF-couplet) on mouse model of acute lung injury (ALI) induced by lipopolysaccharide (LPS) and on RAW264.7 cell inflammation induced by LPS, and investigate the anti-inflammatory mechanism NF-κB signal pathway. Methods ①ALI model: male Kunming mice were divided into blank group, sham-operation group, model group, dexamethasone +LPS group (5 mg/kg, DXM group) and low-dose, mid-dose and high-dose NBE-JF-couplet +LPS groups (3.33, 6.67, 10.01 g/kg, low-dose, mid-dose and high-dose NBE-JF-couplet groups). DXM group was given intraperitoneal injection of DXM once on the 2nd, 4th and 5th d, NBE-JF-couplet groups were orally given corresponding medicinals for 5 d (once a day), and blank group, sham-operation group and model group were given equivalent Tween solution (0.5%). After the last medication for 30 min, except of blank group and sham-operation group, other groups were given intratracheal drip of LPS (100 μL, 5 mg/kg) for establishing ALI model, and sham-operation group was given anesthesia and intratracheal drip of saline solution. All groups were medicated once more after modeling for 5 h, and mice were executed after LPS administration for 6 h. The samples of lung tissue were collected from mice for pathological histomorphological observation. The mRNA expressions of lung NF-κB p65, NF-κB p50 and IκBα were detected by using RT-PCR. The expression of NF-κB p50 protein was detected by using Western blotting assay. ②Inflammatory model of RAW264.7 cells: RAW264.7 cell suspension in logarithmic term was inoculated and cultured for 24 h, the cells were divided into blank group, LPS group (1 μg/mL), DMSO group, DXM group (5 mg/L), high-dose and low-dose NBE-JF-couplet groups (50 mg/L, 25 mg/L), high-dose and low-dose 5-O-methylvisamminol groups (50, 25 mg/L), hesperidin group (50 μg/L) and rosmarinic acid group (5 mg/L). After pretreatment with medicinals for 3 h, all groups, except of blank group, were given LPS (1 mg/L) for 12 h to establish model and absorb supernatant. The content of NO was detected by using Griess method, and levels of IL-6 and TNF-α were detected by using ELISA. The mRNA expressions of NF-κB p65, NF-κB p50 and IκBα were detected by using RT-PCR. Results ①The count of neutrophils decreased in all NBE-JF-couplet groups compared with model group, which was more significant in low-dose NBE-JF-couplet group (P<0.01). The expressions of lung NF-κB p65 mRNA and NF-κB p50 protein were down-regulated in mid-dose and low-dose NBE-JF-couplet group (P<0.01), and expression of NF-κB p50 mRNA was significantly down-regulated in mid-dose NBE-JF-couplet group (P<0.05). The level of IL-6 decreased significantly in all medicated groups compared with model group (P<0.01 or P<0.05). The content of NO decreased significantly in high-dose and low-dose NBE-JF-couplet groups and low-dose 5-O-methylvisamminol groups (P<0.01 or P<0.05). The level of TNF-α had a descending trend in high-dose NBE-JF-couplet group. The mRNA expressions of NF-κB p65, NF-κB p50 and IκBα were down-regulated significantly in low-dose NBE-JF-couplet group (P<0.05 or P<0.01). The mRNA expressions of NF-κB p50 and IκBα were down-regulated significantly in high-dose NBE-JF-couplet group (P<0.05 or P<0.01). The expressions of NF-κB p50 mRNA was down-regulated significantly in high-dose and low-dose 5-O-methylvisamminol groups and hesperidin group (P<0.05). The mRNA expressions of NF-κB p50 and IκBα were down-regulated significantly in rosmarinic acid group (P<0.05). Conclusion The n-butanol extracts from couplet medicinals of Jingjie and Fangfeng has effects against acute inflammation, which is related to inhibition of mRNA and protein expressions of NF-κB p65, NF-κB p50, IκBα and NF-κB p50 in NF-κB signaling pathway, and 5-O-m-ethylvisamminol, hesperidin and rosmarinic acid may be the main material basis.
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