Das Sreemanti, Saha Santu Kumar, De Arnab, Das Durba, Khuda-Bukhsh Anisur Rahman
Cytogenetics and Molecular Biology Laboratory, Department of Zoology, University of Kalyani, India..
Zhong Xi Yi Jie He Xue Bao. 2012 Mar;10(3):337-46. doi: 10.3736/jcim20120314.
To examine to what degree an ultra-highly diluted homeopathic remedy, Arnica Montana 30C (AM-30C), used in the treatment of shock and injury, can modulate the expression of nucleotide excision repair genes in Escherichia coli exposed to ultraviolet (UV) irradiation.
E. coli were cultured to their log phase in a standard Luria-Bertani medium and then exposed to sublethal doses of UV irradiation at 25 and 50 J/m(2) for 22.5 and 45 s, respectively. The UV-exposed bacteria were then supplemented with either AM-30C (drug) or placebo (P-30C). The drug-treated and placebo-treated bacteria were subjected to assay for DNA damage and oxidative stress 90 min after UV exposure. Several protocols like comet assay, gel electrophoresis for DNA ladder and intracellular reactive oxygen species (ROS) generation, and biomarker measurement like superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) were conducted. The mRNA expressions of the excision repair genes like ultraviolet repair uvrA, B and C genes (or also known as excision repair genes) were estimated by reverse transcription-polymerase chain reaction method.
The UV-exposed bacteria showed DNA damage and oxidative stress, as revealed by an increase in ROS generation, and a decrease in SOD, CAT and GSH activities. As compared to placebo, the AM-30C-treated bacteria showed less DNA damage and oxidative stress as manifested by a decrease in ROS generation, and an increase in SOD, CAT and GSH activities. AM-30C also up-regulated the expression of repair genes as compared to the control.
AM-30C helped repair the DNA damage through up-regulation of repair genes and also ameliorated the oxidative stress through the reduction of ROS generation and suitable modulation of anti-oxidative stress enzymes.
研究用于治疗休克和损伤的超高稀释顺势疗法药物山金车30C（AM - 30C）在多大程度上能够调节暴露于紫外线（UV）照射下的大肠杆菌中核苷酸切除修复基因的表达。
将大肠杆菌在标准的Luria - Bertani培养基中培养至对数期，然后分别以25 J/m²和50 J/m²的亚致死剂量紫外线照射22.5秒和45秒。之后，给紫外线照射后的细菌补充AM - 30C（药物）或安慰剂（P - 30C）。紫外线照射90分钟后，对药物处理组和安慰剂处理组的细菌进行DNA损伤和氧化应激检测。进行了几种实验方案，如彗星试验、DNA梯状条带的凝胶电泳和细胞内活性氧（ROS）生成检测，以及超氧化物歧化酶（SOD）、过氧化氢酶（CAT）和还原型谷胱甘肽（GSH）等生物标志物的测量。通过逆转录 - 聚合酶链反应法估计切除修复基因如紫外线修复uvrA、B和C基因（也称为切除修复基因）的mRNA表达。
紫外线照射后的细菌表现出DNA损伤和氧化应激，表现为ROS生成增加，SOD、CAT和GSH活性降低。与安慰剂相比，AM - 30C处理的细菌表现出较少的DNA损伤和氧化应激，表现为ROS生成减少，SOD、CAT和GSH活性增加。与对照组相比，AM - 30C还上调了修复基因的表达。
AM - 30C通过上调修复基因帮助修复DNA损伤，还通过减少ROS生成和适当调节抗氧化应激酶减轻氧化应激。
