Toxicology Evaluation Of Huang-Dai-Pian (HDP) And Realgar

Objective:Huang-Dai Pian(HDP) is a realgar (As4S.4)-containing Chinese medicine for hematologic malignancies. Realgar is the main component but is often mistaken as arsenate for risk assessment. To evaluate true risk of realgar and HDP, acute toxicity and subacute toxicity were examined in mice.Methods:In acute toxicity experiment, mice were orally given HDP and equivalent dose of realgar (20mg As·kg-1). sodium arsenate (10mg As·kg-1). Serum biochemistry, tissue arsenic content, arsenic toxicity-sensitive gene expressions and histopathology were determined. In subacute toxicity experiment, mice were orally given HDP (140mg As·kg-1) and realgar nanoparticle (7mg As·kg-1) for20days. Serum biochemistry and pathology of liver and kidney were determined.Results:In acute toxicity experiment, arsenate increased serum ALT. AST and BUN levels, indicative of liver and kidney injury. Histopathology showed severe damage in arsenate-treated liver and kidneys, while in HDP and realgar-treated animals, these lesions were mild or absent and no elevations in serum biochemistry. Hepatic and renal arsenic contents were dramatically increased after arsenate. but only slightly increased after HDP or realgar. Expressions of arsenic toxicity sensitive genes, namely metallothionein (MT-1and MT-2). heme oxygenase-1(HO-1) and interleukin-1β (IL-1β) were marked increased by arsenate. but only slightly increased by HDP and realgar. In subacute toxicity experiment, there was no overt toxicity in HDP and realgar nanopaticle-treated animals, but BUN was slightly increased and histopathology showed slight injury in HDP animals.Conclusion:HDP and realgar are low toxicity. The use of total arsenic content to evaluate the safety of realgar-containing traditional medicine such as HDP is inappropriate. Objective:To study the anti-tumor effect of Huang-Dai-Picm (HDP) and realgar nanoparticle in vivo.Methods:Human hepatocarcinoma HepG2cells were injected into nude mice to establish tumor-transplant model in vivo. Mice were divided into4groups. The positive control group was given cisplatin1mg·kg-1. the experiment groups were given realgar nanoparticle10mg·kg-1, HDP4400mg·kg-1plus cisplatin1mg·kg-1. Tumor volumes were determined and at the end of the experiment, tumor weight, praxiology evaluation, blood biochemistry, histopathology and related gene expressions were examined. For the skin cancer model, skin adenoma cells were injected into BALB/c mice. Mice were divided into5groups. The positive control group was given cisplatin1mg·kg-1, the experiment groups were given realgar nanoparticle10mg·kg-1. HDP4400mg·kg-1plus cisplatin1mg·kg-1and Liu-Shen-Wan200mg·kg-1plus cisplatin1mg·kg-1. Praxiology evaluationand was determined and at the end of the experiment, tumor weights were examined.Results:HDP plus cisplatin inhibited the growth of hepatocarcinoma. The inhibit rate was52.5%. There was no significant damage to liver and kidney. The Bax gene expression showed no difference from control group, but MT-2transcript levels were increased. HDP plus cisplatin, LSW plus cisplatin and realgar nanoparticle had anti-tumor effects against skin cancer model. The inhibition rates were70.4%.79.8%and45.2%. respectively.Conclusion:HDP plus cisplatin synergistically inhibited the growth of hepatocarcinoma and skin adenoma cell growth in mice. Objective:To examine circadian variations of hepatic antioxidant components, including the Nrf2-pathway, the glutathione (GSH) system, antioxidant enzymes and metallothionein in mouse liver.Methods:Adult KunMing mice were randomly divided into12groups. The male and female were half and half. Mice were housed in light-and temperature-controlled facilities(room temperature22±2℃,8:00am light on and8:00pm light off) for2weeks, and livers were collected every4h during the24h period. Total RNA was isolated, purified, and subjected to real-time RT-PCR analysis. Hepatic GSH content was determined by the enzyme-linked immunity assay.Results:Hepatic mRNA levels of Nrf2. Keap1,Nqol and Gclc were higher in the light-phase than the dark-phase, and were female-predominant. Hepatic GSH presented marked circadian fluctuations, along writh glutathione S-transferases (GST-al, GST-μ. GST-π) and glutathione peroxidase (GPxl). The expressions of GPxl. GST-μ. and GST-π mRNA were also higher in females. Antioxidant enzymes catalase (CAT). cyclooxygenase-2(Cox-2) and heme oxygenase-1(Ho-1) showed circadian rhythms, with higher expressions of Cox-2and CAT in females. Metallothionein. a small non-enzymatic antioxidant protein, showed dramatic circadian variation in males, but higher expression in females. The circadian variations of hepatic antioxidant components resemble the clock gene Brain and Muscle Arnt-like Protein-1(Bmall). albumin site D-binding protein (Dbp). nuclear receptor Rev-Erba (Nr1d1). period protein (Per1and Per2) and cryptochrome1(Cry1). respectively.Conclusions:Circadian variations and gender differences in transcript levels of antioxidant genes exist in mouse liver. which could affect individual responses to oxidative stress at different times of the day.