In a continuing effort to discover more anti-inflammatory medicinal plants in China, the anti-inflammatory activities of 101 extracts from different parts of 84 traditional medicinal plants were evaluated by a panel of in vitro and in vivo assays. Nuclear factor-kappa B (NF-
Inflammation is an adaptive response of body tissues to external challenge or cellular injury. It is generally thought that inflammation is a beneficial host response defense system, but it can become harmful if dysregulated [
Traditional Chinese medicinal plants work well for many diseases and have been used for centuries in China. Its increasing use in recent years is evidence of a public interest in having alternatives to conventional medicine [
Overview of the collected plants used in this investigation.
Number | Latin name | Plant part | Sample location |
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(1) |
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Lushan, Jiangxi |
(2) |
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Zhangjiajie, Hunan |
(3) |
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Shijiazhuang, Hebei |
(4) |
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Chengdu, Sichuan |
(5) |
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Jingzhou, Hubei |
(6) |
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Jingzhou, Hubei |
(7) |
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Lushan, Jiangxi |
(8) |
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Lushan, Jiangxi |
(9) |
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Lushan, Jiangxi |
(10) |
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Lushan, Jiangxi |
(11) |
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Lushan, Jiangxi |
(12) |
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Lushan, Jiangxi |
(13) |
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Lushan, Jiangxi |
(14) |
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Xishuangbanna, Yunnan |
(15) |
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Jingzhou, Hubei |
(16) |
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Yunwushan, Guangdong |
(17) |
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Jingzhou, Hubei |
(18) |
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Shanghai |
(19) |
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Xishuangbanna, Yunnan |
(20) |
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Xishuangbanna, Yunnan |
(21) |
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Yunwushan, Guangdong |
(22) |
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Shanghai |
(23) |
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Xishuangbanna, Yunnan |
(24) |
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Bozhou, Anhui |
(25) |
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Chengdu, Sichuan |
(26) |
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Shijiazhuang, Hebei |
(27) |
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Shijiazhuang, Hebei |
(28) |
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Chengdu, Sichuan |
(29) |
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Chengdu, Sichuan |
(30) |
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Xishuangbanna, Yunnan |
(31) |
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Shanghai |
(32) |
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Chengdu, Sichuan |
(33) |
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Shanghai |
(34) |
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Shanghai |
(35) |
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Shanghai |
(36) |
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Shanghai |
(37) |
|
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Shanghai |
(38) |
|
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Xishuangbanna, Yunnan |
(39) |
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Shanghai |
(40) |
|
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Lushan, Jiangxi |
(41) |
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Chengdu, Sichuan |
(42) |
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Emeishan, Sichuan |
(43) |
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Emeishan, Sichuan |
(44) |
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Shanghai |
(45) |
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Shijiazhuang, Hebei |
(46) |
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Lushan, Jiangxi |
(47) |
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Lushan, Jiangxi |
(48) |
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Lushan, Jiangxi |
(49) |
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Bozhou, Anhui |
(50) |
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Zhangjiajie, Hunan |
(51) |
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Shanghai |
(52) |
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Bozhou, Anhui |
(53) |
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Lushan, Jiangxi |
(54) |
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Lushan, Jiangxi |
(55) |
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Zhangjiajie, Hunan |
(56) |
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Lushan, Jiangxi |
(57) |
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Shanghai |
(58) |
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Shanghai |
(59) |
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Shanghai |
(60) |
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Bozhou, Anhui |
(61) |
|
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Shijiazhuang, Hebei |
(62) |
|
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Shijiazhuang, Hebei |
(63) |
|
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Shanghai |
(64) |
|
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Lushan, Jiangxi |
(65) |
|
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Jinyunshan, Chongqing |
(66) |
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Jingzhou, Hubei |
(67) |
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Bozhou, Anhui |
(68) |
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Chengdu, Sichuan |
(69) |
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Shanghai |
(70) |
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Zhangjiajie, Hunan |
(71) |
|
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Shanghai |
(72) |
|
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Shijiazhuang, Hebei |
(73) |
|
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Yunwushan, Guangdong |
(74) |
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Emeishan, Sichuan |
(75) |
|
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Shanghai |
(76) |
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Shanghai |
(77) |
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Shanghai |
(78) |
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Lushan, Jiangxi |
(79) |
|
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Shanghai |
(80) |
|
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Lushan, Jiangxi |
(81) |
|
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Chengdu, Sichuan |
(82) |
|
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Yunwushan, Guangdong |
(83) |
|
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Jingzhou, Hubei |
(84) |
|
|
Xishuangbanna, Yunnan |
(85) |
|
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Shanghai |
(86) |
|
|
Zhangjiajie, Hunan |
(87) |
|
|
Xishuangbanna, Yunnan |
(88) |
|
|
Yunwushan, Guangdong |
(89) |
|
|
Bozhou, Anhui |
(90) |
|
|
Chengdu, Sichuan |
(91) |
|
|
Lushan, Jiangxi |
(92) |
|
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Lushan, Jiangxi |
(93) |
|
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Lushan, Jiangxi |
(94) |
|
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Lushan, Jiangxi |
(95) |
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Shanghai |
(96) |
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Chengdu, Sichuan |
(97) |
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Bozhou, Anhui |
(98) |
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Bozhou, Anhui |
(99) |
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Bozhou, Anhui |
(100) |
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Xishuangbanna, Yunnan |
(101) |
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|
Zhangjiajie, Hunan |
The plant materials used in this study were collected from different locations of China. The collected species were identified and authenticated by taxonomists from the Second Military Medical University (China). All plant samples were air-dried and finally ground to a fine powder before further processing.
The medicinal plants used in this study were collected at the different areas in China. All voucher specimens have been deposited at the Lab. of Chemistry and Bioactivity of TCM, Shanghai Jiaotong University. The air dried powder of medicinal plants was extracted with 95% ethanol three times (24 h × 3) at room temperature. The extract was evaporated in vacuo to leave a residue extract, which was measured and dissolved in DMSO as the final concentration of 10 mg/mL, respectively.
Male C57BL/6 mice, 6–8 weeks old and weighing approximately 20 g, were purchased from SLACCAS Laboratory Animal Co., Ltd (Shanghai, China). All mice were housed (four to five animals per cage) with a 12/12 h light/dark cycle, with ad libitum access to food and water. The housing, breeding, and animal experiments were in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals, with procedures approved by the Biological Research Ethics Committee of Shanghai Jiao Tong University.
The RAW 264.7 macrophage cell line was obtained from the China Cell Line Bank (Beijing, China). The cells were cultured in DMEM medium supplemented with 10% FBS, penicillin (100 U/mL), and streptomycin (100
The examination of NF-
Cells were plated overnight in 96-well plate (5 × 103/well). Then, the cells were treated with 0.1% DMSO (vehicle control) or the plant extract samples with various concentrations for 24 h. Following another 4 h incubation with 20
RAW 264.7 macrophages were plated in 48-well plate (1.5 × 105/well). After cells became adherent, they were pretreated with the test samples, positive control (parthenolide, 10
RAW 264.7 macrophages were plated in 12-well plate (5 × 105/well). Cells pretreated with the test samples, positive control (parthenolide, 10
Xylene-induced ear edema in mice was initiated according to a previously described method [
The results are expressed as mean ± SEM. Statistical differences were compared with one-way ANOVA.
In a primary screening, 101 different extracts from 84 plants were assessed for their potential to inhibit TNF-
Inhibitory effects of the plant extracts (50
In order to exclude the potential influence of cytotoxicity, the effects of these 22 effective plant extracts at the concentrations of 50, 25, and 12.5
Cytotoxic effects of the extracts in the MTT assay (Hela cells and Raw 264.7 cells).
Number | Latin name | Hela cells | Raw 264.7 cells | ||||
---|---|---|---|---|---|---|---|
Viability at 12.5 |
Viability at 25 |
Viability at 50 |
Viability at 12.5 |
Viability at 25 |
Viability at 50 | ||
1 |
|
105.0 ± 1.2 | 107.0 ± 5.2 | 110.5 ± 2.0 | 122.1 ± 2.4 | 129.6 ± 0.8 | 143.6 ± 2.5 |
2 |
|
87.4 ± 2.5 | 86.0 ± 1.9 | 75.8 ± 1.4 | 110.6 ± 1.3 | 99.7 ± 1.6 | 81.3 ± 1.9 |
6 |
|
103.8 ± 0.3 | 127.9 ± 3.3 | 119.4 ± 3.7 | 55.1 ± 2.9 | 55.8 ± 4.6 | 49.2 ± 3.9 |
7 |
|
105.4 ± 2.0 | 120.8 ± 2.6 | 127.8 ± 4.6 | 130.8 ± 0.8 | 130.0 ± 2.2 | 134.2 ± 2.4 |
9 |
|
61.7 ± 1.3 | 49.1 ± 1.6 | 32.2 ± 1.2 | NT | NT | NT |
26 |
|
110.9 ± 0.3 | 126.7 ± 2.0 | 112.3 ± 3.8 | 123.0 ± 1.3 | 126.0 ± 3.0 | 145.3 ± 1.8 |
27 |
|
130.6 ± 3.3 | 127.9 ± 4.6 | 12.4 ± 0.8 | 132.2 ± 7.0 | 112.0 ± 2.1 | 12.1 ± 0.9 |
42 |
|
25.3 ± 1.8 | 09.8 ± 0.9 | 10.3 ± 1.0 | NT | NT | NT |
43 |
|
13.9 ± 1.9 | 08.9 ± 0.6 | 7.5 ± 0.4 | NT | NT | NT |
46 |
|
84.5 ± 3.5 | 92.8 ± 1.5 | 85.0 ± 2.3 | 121.2 ± 4.1 | 112.0 ± 0.8 | 113.2 ± 0.9 |
50 |
|
87.0 ± 1.7 | 43.5 ± 2.4 | 8.4 ± 0.2 | NT | NT | NT |
55 |
|
88.6 ± 0.8 | 85.9 ± 1.4 | 81.8 ± 4.5 | 80.5 ± 3.9 | 75.0 ± 4.8 | 68.0 ± 3.0 |
62 |
|
121.8 ± 1.8 | 77.9 ± 3.0 | 71.6 ± 4.6 | 118.2 ± 0.5 | 119.9 ± 3.2 | 101.6 ± 4.7 |
77 |
|
128.2 ± 1.9 | 127.6 ± 1.4 | 129.1 ± 2.5 | 151.4 ± 0.4 | 179.2 ± 2.0 | 170.5 ± 3.9 |
80 |
|
103.4 ± 7.1 | 84.0 ± 3.8 | 64.9 ± 3.5 | 142.4 ± 0.5 | 141.9 ± 1.4 | 136.3 ± 1.5 |
85 |
|
81.9 ± 1.6 | 69.6 ± 2.6 | 47.7 ± 2.8 | NT | NT | NT |
86 |
|
112.1 ± 2.5 | 93.0 ± 3.6 | 56.1 ± 0.9 | 88.4 ± 2.3 | 89.8 ± 0.6 | 76.9 ± 3.4 |
87 |
|
118.9 ± 3.1 | 79.5 ± 1.2 | 65.5 ± 3.6 | 94.0 ± 1.0 | 114.2 ± 4.4 | 104.5 ± 0.3 |
88 |
|
36.3 ± 0.3 | 13.0 ± 0.7 | 10.0 ± 1.0 | NT | NT | NT |
89 |
|
10.6 ± 1.4 | 7.9 ± 0.4 | 5.5 ± 0.1 | NT | NT | NT |
90 |
|
30.3 ± 0.2 | 6.9 ± 0.5 | 7.3 ± 0.8 | NT | NT | NT |
101 |
|
63.0 ± 0.8 | 66.7 ± 3.4 | 51.3 ± 1.0 | NT | NT | NT |
Values are means ± standard deviations. NT: not tested.
The inhibitory effects of the nine plant extracts on NO production in LPS-stimulated RAW 264.7 cells were assessed. As shown in Figure
Inhibitory effect of plant extracts (50
The effects of the above nine extracts active in both NF-
Inhibitory effects of plant extracts (50
The effect of
Traditional Chinese medicinal plants play a significant role in drug discovery and development. Many have provided the foundation for modern pharmaceuticals and drug leads [
NF-
Macrophage activation by LPS leads to the phosphorylation, ubiquitination, and subsequent degradation of the inhibitory
In summary, the present study demonstrates that extracts from nine medicinal plants exhibit properties of anti-inflammatory activity by either suppressing NO production or the major proinflammatory cytokines TNF-
The authors declare that there are no conflicts of interest regarding the publication of this paper.
Chao Fan and Hui-zi Jin have contributed equally to this work.
This work was funded by National Science Foundation of China (81402806). This work was also supported in part by grants from the National Natural Science Foundation of China (Grant 31470856), the Science and Technology Development Fund of Macau (FDCT 026/2016/A1 and 072/2015/A2), and the University of Macau (CPG2015-00018-ICMS).