Shangqingyin (SQY), made from Fructus Chebulae, Frangipani, Reed Rhizome, Mesona, Patchouli, Liquorice, has been a famous herbal tea in Lingnan area in China since Tang dynasty. It is believed to be beneficial for human health, although experimental data are scarce until now. In the present study, we demonstrated that oral administration of SQY herbal tea significantly decreased the weights of granuloma in rats induced by cotton pellet, mitigated the paw edema induced by dextran in rats and alleviated the ear edema induced by dimethylbenzene in mice. These results indicated a potential anti-inflammatory effect of SQY, which preliminarily provide a scientific basis for its beneficial effects.
香云(SQY),抗炎作用,大鼠肉芽肿测试,大鼠爪水肿测试,小鼠耳朵水肿测试
炎症是各种刺激(例如创伤,感染和有毒化学物质)引起的组织损伤的生理和免疫反应的正常结果[1,2]。它在肿瘤发育,心血管疾病,糖尿病,帕金森氏病,等。Therefore, it’s of great value to look for anti-inflammation drugs. In Tang dynasty of China, people in Lingnan area in China often drank myrobalan soup to keep healthy. It was prepared by decocting fresh myrobalan (Fructus Chebulae) with Liquorice, tasting slightly sweet. In Ayurveda, Fructus Chebulae was commonly used as cardiotonic, digestive, diuretic, antitussive, antidiabetic, and laxative [3,4]. Meanwhile, Fructus Chebulae was previously reported to have anticancer [5], antioxidant [6,7], anti-hyperglycemic [8], antimutagenic [9], anti-ulcer [10], hepatoprotective [11], and cardio-protective activities [12]. As time goes by, people combined Fructus Chebulae with some other traditional Chinese medicine, such as Frangipani, Reed Rhizome, Mesona, Patchouli, Liquorice, to make a herbal tea named SQY. In the present study, we investigated the anti-inflammatory effects of SQY herbal tea by employing three inflammation models including cotton pellet induced rat granuloma, dextran-induced rat paw edema and dimethylbenzene-induced ear edema.
Drugs and major reagents
SQY(总净重:4.526公斤)由Xiangxue Pharmaceutical Co.,Ltd。(中国广州)提供。还使用了地塞米松磷酸钠注射(DXM)(批次:51504092)是从Tianjin King Group Hubeitianyao Pharmaceutical Co.,Ltd。(中国Tianjin)购买的。
动物
SPF SD rats (170-190 g, male) and Kunming mice (18-22 g, male) were obtained from Guangdong Medical Laboratory Animal Center (License No: SCXK 2013-0002). All animals were acclimatized housing environment with 23 ± 2℃ and 12:12 h light-dark cycle for one week before the experiments. They were maintained on standard chow pellet and water ad libitum. All animal care and experimental procedures were approved by the Laboratory Animal Ethics Committee of Jinan University, and were in accordance with the National Institute of Health’s Guide for the Care and Use of Laboratory Animals (the 7th美国版)。
棉沉淀诱导大鼠肉芽肿测试
This test was performed according to the report of Santos and Rao [13]. SD rats were divided into six groups with ten each, including control group, dexamethasone group (DXM,I.P.,5 mL/kg)和四个SQY治疗组的剂量。根据人的日常饮酒量(10.33 mL/kg),分别用25.83 mL/kg,51.65 mL/kg,103.3 mL/kg和309.9 mL/kg的大鼠使用25.83 mL/kg,51.65 mL/kg,103.3 mL/kg和309.9 ml/kg的大鼠大鼠。对照组中的大鼠用等量的纯净水口服处理。这些药物每天每天管理一次30天。在23上th药物治疗的日期,将棉组颗粒(23±1 mg)手术植入小鼠右腹股沟的皮肤下。然后将伤口缝合,并从麻醉中恢复后单独笼罩动物。在30th一天,处死大鼠,并将颗粒与肉芽肿一起去除。除去外部组织,然后将沉淀组织放入板中,并在烤箱中用60℃干燥1小时,直到重量保持恒定为止。最后,沉淀组织的重量为重量,并表示为颗粒肉芽组织的干重与原始小棉球的相应重量之间的差异。
Dextran-induced rat paw edema test
该测试是根据Ribeiro Na的报告进行的等。[14]动物组和药物治疗与棉沉淀诱导的大鼠肉芽肿测试相同。在30thday, acute inflammation was produced by subplantar injection of 0.1 mL of freshly prepared 1% (w/v) dextran in normal saline into the right hind paws of rats. One hour after daily drug treatment, paw volume was measured plethysmometrically using a paw edema calcimeter at 0, 1, 2, 4 and 6 hours after dextran injection. The same region was measured three times and the average was taken. The edema volumes were the difference between the paw volumes measured at different time and the paw volumes at zero hour. Calculate the paw edema rate each period of time. Paw edema rate was expressed as the paw edema volumes divide the paw volumes before the inflammatory agent effect.
Dimethylbenzene-induced mouse ear edema test
Mouse ear edema test was referenced to Zhang YB等。和李QZ等。(15、16)。所有小鼠随机分为7格ups (n=12), also including control group, DXM group and SQY-treatment groups. The rats in control group were treated with equivalent volume of purified water orally. The rats in SQY-treatment groups were gavaged with 25.83, 51.65, 103.3, 155.0 and 309.9 mL/kg SQY, respectively. In DXM group, mice were treated the same as described above. Each group should be dosed once daily for 30 days. On the 30th每天的药物治疗后一小时,将二甲基苯(20μL/耳朵)局部应用于小鼠的右耳。没有任何应用的左耳是对照组。30分钟后,用宫颈脱位处置小鼠,然后用冲孔(直径为9毫米)获得两只耳朵的耳朵活检并称重。通过从右侧减去左耳的重量来确定耳朵水肿值。耳水水肿率表示为耳水水肿值,将对照耳片切片的重量分开。
Statistical analysis
数据表示为平均值±SEM,并使用GraphPad Prism 6软件通过Student's t检验进行分析,P <0.05被认为具有统计学意义。
棉沉淀诱导大鼠肉芽肿测试的结果
如图1所示,在25.83和51.65 mL/kg的剂量下,SQY显着降低了大鼠棉花颗粒诱导的肉芽瘤重量(p <0.01)。Nevertheless, comparing with control group, SQY (103.3 and 309.9 mL/kg) treatment didn’t strikingly influence the granuloma weights (338.6 ± 23.09 in control group vs. 280.8 ± 27.24 in 103.3 mL/kg SQY group and 270.7 ± 27.2 in 309.9 mL/kg SQY group).
![](//m.script-one.com/img/IFNM-4-174-g001.gif)
2021版权燕麦。所有权利保留
Figure 1.SQY在棉沉淀诱导的大鼠肉芽肿试验中的抗炎作用[颗粒颗粒组织是重量和不同组的肉芽肿的净体重(对照,DXM,25.83 ml/kg,51.65 ml/kg,103.3 ml/kg和309.999.99.999.99.9ML/kg的SQY处理)表示为颗粒肉芽组织的干重与原始小棉球的相应重量之间的差异。与对照组相比,** p <0.01,*** p <0.001]
葡聚糖诱导的大鼠爪水肿测试的结果
In the Figure 2A, SQY treatment had no obvious effect on the paw edema rate at 1 h after dextran-administration. However, the SQY-treatment groups had significant differences compared with the control group at 2 h, 4 h and 6 h after dextran injection (Figures 2B-2D). This data indicated that SQY exerted anti-inflammation effects in dextran-induced rat paw edema test, but the initial effective time of the anti-inflammatory effect of SQY was slightly slowly when compared with DXM.
![](//m.script-one.com/img/IFNM-4-174-g002.gif)
Figure 2.Anti-inflammation effects of SQY in dextran-induced rat paw edema test [The paw edema volumes were the paw volumes’ difference measured between at different time and at zero hour by a paw edema calcimeter. Paw edema rate was expressed as the paw edema volumes divide the paw volumes before the inflammatory agent effect. Paw edema rates were determined at 1 h (A), 2 h (B), 4 h (C), 6 h (D), and the time point changes is indicated in (E). Compared with control group, *P < 0.05, **P < 0.01, ***P < 0.001]
结果of dimethylbenzene-induced mouse ear edema test
如图3所示,在51.65、103.3、155.0和309.9 mL/kg的剂量下,SQY显着降低了由二甲基苯引起的耳水肿率,尽管SQY在25.83 mL/kg的剂量下没有显着影响。这些结果表明,在较高剂量下,对二甲基苯诱导的耳水肿有抗炎作用。
Figure 3.SQY在二甲基苯诱导的小鼠耳水肿测试中的抗炎作用[耳水水肿值是通过从右耳的右右耳(20μl/Ear)中减去左耳的重量来确定的。耳水水肿率表示为耳水水肿值,将对照耳片切片的重量分开。与对照组相比,** p <0.01,*** p <0.001]。
在本研究中,我们利用了三个体内模型,包括棉沉淀诱导的大鼠肉芽肿测试和右旋烷诱导的大鼠爪水肿测试,以及二甲基苯诱导的小鼠耳朵水肿测试,以检查SQY的抗炎作用。棉沉淀引起的肉芽肿的形成是慢性炎症反应的典型症状。该过程已被广泛用于评估慢性炎症的渗出性和增殖成分,因为棉花颗粒的干重与肉芽肿组织的量密切相关[17]。因此,我们首先采用棉沉淀诱导的大鼠肉芽肿测试来研究SQY的抗炎作用,并在51.65、103.3 mL/kg的剂量下显着降低了肉芽肿的重量,并且对DXM治疗的疗效封闭了。LO等。and Calixto [18-19] reported that edema produced by subplantar injection of dextran in animals is characterized by a rapid increase in the paw edema and spontaneous decrease after 30 min, with histamine and serotonin being the main mediators. In our study, a significant anti-inflammatory effect of SQY were also observed in the dextran-induced rat paw edema test at the dosages of 25.83 and 51.65 mL/kg, other than at higher dosages 103.3 and 309.9 mL/kg. We speculate that SQY may cause allergy at high dosages, due to Frangipani in SQY have been reported to induce such adverse effects [20]. In order to further evaluate the anti-inflammatory activity of the extract, the dimethylbenzene-induced ear edema test was also employed. In the process of inflammation, activated neutrophils release mediators such as platelet-activating factor and lysozyme, which can lead to vasodilatation and increase vascular permeability [21]. In the present study, it was also shown that middle concentration SQY significantly inhibited ear edema of mice. In the dimethylbenzene-induced mouse ear edema test, SQY had obvious anti-inflammatory effects at the dosages of 51.65, 155.0, 309.9 and 103.3 mL/kg while 25.83 mL/kg showed no significant effect. By analyzing the results from above tests, it can be seen that the effective dosages of SQY in different tests were distinct. It may be due to the species differences and model distinction.
SQY Fructu制成s Chebulae, Patchouli and Liquorice, which have been reported to possess anti-inflammatory effects. Lee等。demonstrated Fructus Chebulae inhibited the production of inflammatory mediators and reduced nitric oxide (NO) production, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) expression in RAW 264.7 [22]. Yoon等。reported that the aqueous extract of Patchouli was proved to have anti-allergic and anti-inflammatory effects by reduced intracellular calcium levels and the activation of NF-κB and p38 MAPK. [23]. What’s more, Menegazzi等。proved that glycyrrhizin, a major active constituent of Liquorice root, had anti-inflammatory and anti-viral activities [24]. Thus, it can be inferred that the effects of SQY on inflammation might be associated with these components. Nevertheless, what’s the exact mechanism needs more investigation in the future study.
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