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茶树(L.)在胶原诱导性关节炎模型中的体内抗关节炎功效。

In vivo anti-arthritic efficacy of Camellia sinensis (L.) in collagen induced arthritis model.

作者信息

Tanwar Ankit, Chawla Raman, Ansari Md Meraj, Thakur Pallavi, Chakotiya Ankita Singh, Goel Rajeev, Ojha Himanshu, Asif M, Basu Mitra, Arora Rajesh, Khan Haider Ali

机构信息

Heavy Metal and Clinical Toxicology Laboratory, Department of Medical Elementology and Toxicology, Hamdard University, Hamdard Nagar, New Delhi, 110062, India; Division of CBRN Defence, Institute of Nuclear Medicine and Allied Sciences, Delhi, India.

Division of CBRN Defence, Institute of Nuclear Medicine and Allied Sciences, Delhi, India.

出版信息

Biomed Pharmacother. 2017 Mar;87:92-101. doi: 10.1016/j.biopha.2016.12.089. Epub 2016 Dec 31.

DOI:10.1016/j.biopha.2016.12.089
PMID:28049097
Abstract

BACKGROUND

Rheumatoid arthritis (RA), an autoimmune inflammatory disorder with synovial hyperplasia, destruction of cartilage, bone damage is often associated with risk of infections. Such risk could be attributed towards usage of immunosuppressive agents. Thus, the present study was undertaken to evaluate the anti-arthritic efficacy of aquo-alcoholic extract of Camellia sinensis (L.).

MATERIAL AND METHODS

Dried leaves of Camellia sinensis (L.) or Cs were filtered and extracted in 1:1 aqueous: ethanol by Soxhlet apparatus followed by lyophilization and spray drying to develop amorphous powder. Four different oral doses (50, 100, 200, 400mg/kg/body wt.) of aquo-alcoholic extract were evaluated for anti-edematogenic effect in collagen induced arthritis model. The selected anti-arthritic doses of Cs were evaluated for the oxidative stress markers like Glutathione [5-5'dithio-bis-2-nitrobenzoicacid (DTNB)], Superoxide dismutase [Epinephrine], Catalase [Hydrogen peroxide], Lipid peroxidation [Thiobarbituric acid reactive substance (TBARS)], Nitric oxide [Griess reagents:Nitrobluetetrazolium], Articular elastase [N-methoxysuccinyl-Ala-Ala-Pro- Val p-nitroanilide] in joints followed by haematological evaluation including RBC, WBC, Haemoglobin, platelets and haematocrit. To validate these biochemical changes, the radiological and histopathological (Haematoxylin & Eosin) evaluation was also conducted.

RESULTS

The selected anti-arthritic dose of Cs i.e. 400mg/kg/body wt. (∼60% anti-arthritic efficacy on 35th day) could be attributed towards significant (p<0.05) increase in the levels of enzymatic (Superoxide dismutase and Catalase) and non-enzymatic (Glutathione) antioxidants by 34%, 59% and 50% respectively. Simultaneously, the significant (p<0.05) reduction of lipid peroxides, nitrite radical and elastase activity by 32%, 45% & 32% respectively as compare to control indicated overall decrease in oxidative stress. Haematological evaluation revealed restoration of RBC, WBC and platelets level in treatment group. The confirmatory analysis utilizing radiological and histological assessment showed alleviation of joint deformity, tissue swelling, pannus formation and neutrophils infiltration in treatment group as compared to collagen induced arthritis.

CONCLUSION

The analysis showed that Cs can play an effective role in reduction of oxidative stress by modulating levels of antioxidants, reducing levels of free radicals while restoring normal haematopoietic cascade as observed in collagen induced arthritis model. Thus, the cumulative dose impact of 400mg/kg body wt., over a period of 14days also found extremely effective in terms of safeguarding their structural conformity against such auto-immune disorder.

摘要

背景

类风湿性关节炎(RA)是一种自身免疫性炎症性疾病,伴有滑膜增生、软骨破坏和骨损伤,常与感染风险相关。这种风险可能归因于免疫抑制剂的使用。因此,本研究旨在评估茶树(L.)水醇提取物的抗关节炎疗效。

材料与方法

将茶树(L.)的干燥叶片或Cs进行过滤,并用索氏提取器以1:1的水:乙醇比例进行提取,然后进行冻干和喷雾干燥以制成无定形粉末。在胶原诱导的关节炎模型中评估了四种不同口服剂量(50、100、200、400mg/kg体重)的水醇提取物的抗水肿作用。对选定的Cs抗关节炎剂量进行了氧化应激标志物的评估,如谷胱甘肽[5-5'-二硫代双-2-硝基苯甲酸(DTNB)]、超氧化物歧化酶[肾上腺素]、过氧化氢酶[过氧化氢]、脂质过氧化[硫代巴比妥酸反应性物质(TBARS)]、一氧化氮[格里斯试剂:硝基蓝四唑]、关节弹性蛋白酶[N-甲氧基琥珀酰-Ala-Ala-Pro-Val对硝基苯胺],随后进行血液学评估,包括红细胞、白细胞、血红蛋白、血小板和血细胞比容。为了验证这些生化变化,还进行了放射学和组织病理学(苏木精和伊红)评估。

结果

选定Cs的抗关节炎剂量即400mg/kg体重(在第35天约有60%的抗关节炎疗效)可归因于酶促(超氧化物歧化酶和过氧化氢酶)和非酶促(谷胱甘肽)抗氧化剂水平分别显著(p<0.05)增加34%、59%和50%。同时,与对照组相比,脂质过氧化物、亚硝酸盐自由基和弹性蛋白酶活性分别显著(p<0.05)降低32%、45%和32%,表明氧化应激总体降低。血液学评估显示治疗组红细胞、白细胞和血小板水平恢复。利用放射学和组织学评估的验证分析表明,与胶原诱导的关节炎相比,治疗组关节畸形、组织肿胀、血管翳形成和中性粒细胞浸润有所减轻。

结论

分析表明,Cs可以通过调节抗氧化剂水平、降低自由基水平,同时恢复胶原诱导的关节炎模型中观察到的正常造血级联反应,在减轻氧化应激方面发挥有效作用。因此,在14天内400mg/kg体重的累积剂量影响在保护其结构完整性免受这种自身免疫性疾病影响方面也被发现极其有效。

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