Simultaneous determination of acetaminophen and oxycodone in human plasma by LC–MS/MS and its application to a pharmacokinetic study

A simple and rapid liquid chromatography-tandem mass spectrometry （LC-MS/MS） method was de- veloped and validated for simultaneous determination of acetaminophen and oxycodone in human plasma. Acetaminophen-d4 and oxycodone-d3 were used as internal standards. The challenge en- countered in the method development that the high plasma concentration level of acetaminophen made the MS response saturated while the desired lower limit of quantification （LLOQ,） for oxycodone was hard to reach was well solved. The analytes were extracted by protein precipitation using acetonitrile. The matrix effect of the analytes was avoided by chromatographic separation using a hydrophilic C18 column coupled with gradient elution. Multiple reaction monitoring in positive ion mode was performed on tandem mass spectrometer employing electrospray ion source. The calibration curves were linear over the concentration ranges of 40.0-8000 ng/mL and 0.200-40.0 ng/mL for acetaminophen and oxycodone, respectively. This method, which could contribute to high throughput analysis and better clinical drug monitoring, was successfully applied to a pharmacokinetic study in healthy Chinese volunteers.

Trends in oxycodone and oxycodone-containing analgesics administration for back pain in emergency departments in the USA(2007–2018)

BACKGROUND:To describe trends in oxycodone and oxycodone-containing analgesic prescribing for the treatment of back pain among adults in emergency departments(EDs) in the USA from 2007 to 2018.METHODS:Data were gathered from the National Hospital Ambulatory Medical Care Survey(NHAMCS) from 2007 to 2018.The study population included individuals of all ages presenting to USA EDs.The NHAMCS reasons for visit and oxycodone drug ID codes were used to isolate patients with back pain.The main outcome was the proportion of oxycodone and oxycodone-containing analgesics prescribed for back pain in the EDs over the specified time period.RESULTS:There was a relative decrease in the overall administration of oxycodone for back pain in the EDs by 62.3% from 2007(244,000 visits) to 2018(92,000 visits).The proportion of ED patients prescribed with oxycodone-containing analgesics for back pain increased among patients aged 45 years and older(from 43.8% to 57.6%),female patients(from 54.5% to 62.0%),black patients(from 22.5% to 30.4%),and Hispanic/Latino patients(from 9.4% to 19.6%).Oxycodone/acetaminophen was most prescribed and accounted for 90.2% of all oxycodone-containing analgesics in 2007,with a decrease to 68.5% in 2018.Pure oxycodone was the second most prescribed medication,accounting for 6.1% in 2007 and 31.5% in 2018.CONCLUSION:The overall number of oxycodone-containing analgesics decreased significantly from 2007 to 2018.However,that number trended upward in 45-year-old and older,female,black,or Hispanic/Latino patients from 2007 to 2018.The total amount of pure oxycodone increased significantly from 2007 to 2008.

Efficacy and safety of controlled-release oxycodone/naloxone versus controlled-release oxycodone in Korean patients with cancer-related pain: a randomized controlled trial

Background: Controlled-release oxycodone/naloxone(OXN-CR) maintains the effect of opioid-induced analgesia through oxycodone while reducing the occurrence rate of opioid-induced constipation through naloxone. The present study was designed to assess the non-inferiority of OXN-CR to controlled-release oxycodone(OX-CR) for the control of cancer-related pain in Korean patients.Methods: In this randomized, open-labeled, parallel-group, phase IV study, we enrolled patients aged 20 years or older with moderate to severe cancer-related pain [numeric rating scale(NRS) pain score ≥4] from seven Korean oncology/hematology centers. Patients in the intention-to-treat(ITT) population were randomized(1:1) to OXNCR or OX-CR groups. OXN-CR was administered starting at 20 mg/10 mg per day and up-titrated to a maximum of80 mg/40 mg per day for 4 weeks, and OX-CR was administered starting at 20 mg/day and up-titrated to a maximum of 80 mg/day for 4 weeks.The primary efficacy endpoint was the change in NRS pain score from baseline to week4, with non-inferiority margin of-1.5. Secondary endpoints included analgesic rescue medication intake, patientreported change in bowel habits, laxative intake, quality of life(QoL), and safety assessments.Results: Of the ITT population comprising 128 patients, 7 with missing primary efficacy data and 4 who violated the eligibility criteria were excluded from the efficacy analysis. At week 4, the mean change in NRS pain scores was not significantly different between the OXN-CR group(n = 58) and the OX-CR group(n = 59)(-1.586 vs.-1.559,P = 0.948). The lower limit of the one-sided 95% confidence interval(-0.776 to 0.830) for the difference exceeded the non-inferiority margin(P < 0.001). The OXN-CR and OX-CR groups did not differ significantly in terms of analgesic rescue medication intake, change in bowel habits, laxative intake, QoL, and safety assessments.Conclusions: OXN-CR was non-inferior to OX-CR in terms of pain reduction after 4 weeks of treatment and had a similar safety profile. Studies in larger populations of Korean patients with cancer-related pain are needed to further investigate the effectiveness of OXN-CR for long-term pain control and constipation alleviation.Trial registration ClinicalTrials.gov NCT01313780, registered March 8。

Advances in the clinical application of oxycodone in the perioperative period

To review the research progress of pure opioid receptor agonist oxycodone.The research progress of oxycodone in terms of pharmacokinetics,pharmacodynamics,adverse reactions,clinical application,combined medication and new progress in clinical application was summarized by referring to the literature.Oxycodone is a semi-synthetic thebaine derivative of opioid alkaloids,and is a pure opioidμandκreceptor agonist.The main action sites are the central nervous system and visceral smooth muscle.Due to its advantages of low adverse reactions,good analgesic effects,and a wide range of safe doses,the drug has been widely used in the control of acute and chronic postoperative pain,as well as malignant and non-malignant pain.Since the end of the 20^(th) century,researchers have begun to formulate antipyretic analgesics,opioid receptor agonists,opioid receptor antagonists,dopamine receptor antagonists and other drugs with oxycodone in different proportions to enhance the analgesic effect.At the same time,it can reduce the dosage of oxycodone and reduce its adverse reactions,so as to achieve the purpose of limiting opioid abuse.With the continuous research on the efficacy and safety of oxycodone in the perioperative period at home and abroad,oxycodone has become the only dual-opioid potent analgesic that can be used in clinical work.

Effect of dexmedetomidine combined with oxycodone on systemic stress response in recovery of gynecological laparoscopic operation

Objective:To study the effect of dexmedetomidine combined with oxycodone on systemic stress response in recovery period of gynecological laparoscopic operation.Methods:A total of 86 patients who received laparoscopic operation in the Second Affiliated Hospital of Kunming Medical University between June 2014 and December 2016 were selected and randomly divided into the dexmedetomidine combined with oxycodone group (DO group) and control group (C group). Before anesthesia induction (T1), at the end of the surgery and before micro pump injection of dexmedetomidine and oxycodone hydrochloride (T2) and in recovery period (T3), serum levels of pituitary-target gland axis-related hormones, vascular activity-related hormones and oxidative stress-related molecules were determined.Results: At T1 and T2, serum TSH, T3, T4, ACTH, Cor, NE, E, ADH, AT-II, ROS, MDA, SOD and HO-1 levels of DO group were not significantly different from those of C group;at T3, serum TSH, T3, T4, ACTH, Cor, NE, E, ADH, AT-II, ROS and MDA levels of DO group were significantly lower than those of C group while SOD and HO-1 levels were significantly higher than those of C group.Conclusions: Dexmedetomidine combined with oxycodone can inhibit the systemic stress response in recovery of gynecological laparoscopic operation.

Comparison of the Efficacy of Oral Oxycodone and Oral Codeine in the Treatment of Post-Craniotomy Pain—A Randomized, Double-Blind Trial

Background: Post-craniotomy pain has been reported to be moderate to severe. Management of post-craniotomy pain is often inadequate, yet limited by the side effects of opioids. We aim to find out the efficacy of oral oxycodone as compared to oral codeine for the treatment of post-craniotomy pain in our institution. Methods: A randomized, double-blinded controlled trial was used to evaluate the efficacy of oral oxycodone versus oral codeine. 40 patients were randomized to the control group of codeine (n = 20) or the experimental group receiving oxycodone (n = 20) in addition to regular oral paracetamol for both groups of patients. Results: There was no difference in the visual analogue scale scores at 24 hours (2.78 versus 1.85, p = 0.11) or side effects in the oxycodone group compared with the codeine group. Conclusions: Oral oxycodone had similar efficacy as oral codeine in the management of post-craniotomy pain.

Acura制药公司oxycodone口服速释片

羟二氢可待因酮（0xycodone）片剂——OxyADF2期临床试验显示该制剂具有潜在的戒瘾作用。

Development of controlled release bi-layered tablets containing oxycodone hydrochloride

Oxycodone hydrochloride is a semi-synthetic opioid agonist that provides very effective relief for moderate to severe pain in cancer and post-operative patients. Controlled release oxycodone formulations have been studied to enhance the therapeutic effect by providing constant release over the whole dosing interval and improve patient’s convenience by reducing the frequency of administration as well.

Comparison of Efficacy and Safety of Oxycodone Versus Fentanyl for Intravenous Patient-Controlled Analgesia in Postoperative Pain Management:A Systematic Review and Meta-Analysis

Backgroud:Intravenous opioid patient-controlled analgesia(IV-PCA)has been suggested as an effective method in postoperative pain management.There are several randomized controlled trials(RCTs)of comparison of oxycodone and fentanyl for IV-PCA in surgical patients.The purpose of this study was to perform a meta-analysis to compare the efficacy and safety of oxycodone and fentanyl for IV-PCA in surgical patients from current data.Methods:The RCTs of oxycodone versus fentanyl for IV-PCA were gathered from PubMed,Embase,Cochrane library,CNKI and VIP data.After data extraction and quality assessment of the included RCTs,the RevMan 5.3 software was applied for meta-analysis of numerical rating scale(NRS)scores,accumulated IV-PCA consumption of oxycodone and fentanyl,patient satisfaction,postoperative nausea and vomiting(PONV),and other adverse events(AEs).Results:Results reported from eight RCTs involving 600 patients are included in the meta-analysis.The NRS score at rest and upon movement of group oxycodone was significantly lower than that of group fentanyl(WMD=-3.85,95%CI-4.93^-2.76,P<0.00001;WMD=-4.31,95%CI-5.79^-2.84,P<0.00001);however,the incidence of PONV and dizziness was obviously increased in group oxycodone than in group fentanyl(OR=2.41,95%CI 1.60~3.63,P<0.0001;OR=3.69,95%CI 2.17~6.26,P<0.00001).Accumulated IV-PCA consumption in group oxycodone was less than in group fentanyl overall the 48 hours postoperatively(WMD=-12.11,95%CI-18.42^-5.80,P=0.0002).There was no significant difference in patient satisfaction between oxycodone and fentanyl(OR=0.73,95%CI 0.11~5.04,P=0.75).Conclusion:According to the evidence,this meta-analysis suggest that oxycodone for IV-PCA is superior to fentanyl in postoperative pain relief,whereas the higher incidence of PONV and dizziness was accompanied with oxycodone.Further large-scale,prospective,observational studies are needed to summarize and analyse the data to draw a fair conclusion.

Effects of oxycodone and fentanyl patient-controlled intravenous analgesia on pain, immune response and stress response after laparoscopic surgery

Objective:To study the effects of oxycodone and fentanyl patient-controlled intravenous analgesia on pain, immune response and stress response after laparoscopic surgery.Methods:Patients undergoing laparoscopic surgery in Xianning Central Hospital between June 2015 and February 2017 were selected and randomly divided into oxycodone group and fentanyl group who received postoperative oxycodone and fentanyl patient-controlled intravenous analgesia respectively. 3 d after surgery and 5 d after surgery, the serum contents of pain-related transmitters, immune indexes, stress-related molecules as well as peripheral blood contents of immune cells were measured.Results: 3 d after surgery and 5 d after surgery, CRP, TNF-α, IL-8, sICAM-1, YKL-40, Cor, C-P, FT3, FT4 and HO-1 contents in serum of oxycodone group were significantly lower than those of fentanyl group whereas CD3+CD4+T cell and CD3+CD8+T cell contents in peripheral blood as well as C3 and C4 contents in serum were significantly higher than those of fentanyl group.Conclusion:oxycodone patient-controlled intravenous analgesia after laparoscopic surgery is better than fentanyl and can reduce the pain degree, inhibit the stress response and improve the immune response.

Clinical observation on treatment of cancer pain with TCM oriented drugs combined with oxycodone sustained-release tablets and nimesulide sustained-release tablets

Objective： To study the effect of the transdermal preparation of traditional Chinese medicine in treating cancer pain. Methods： From October 2016 to January 2018, 126 patients with cancer pain were enrolled and divided into 4 groups, 39 patients in group A （directed TCM permeation）, 26 patients in group B （oxycodone sustained-release tablets）, 32 patients in group C （Chinese medicine directed drug penetration ＋ oxycodone sustained-release tablets）, and 29 patients group D （Chinese medicine directed drug penetration ＋ oxycodone sustained-release tablets ＋ nimesulide sustained release tablets）, according to KPS scores. Results： Transdermal preparations of traditional Chinese medicine can significantly alleviate cancer pain. For the treatment of moderate to severe cancer pain, the Chinese medicine transdermal preparation can reduce the dosage of oxycodone sustained-release tablets. At the same time, the patient＇s KPS and NRS scores were significantly reduced. Moreover, the transdermal preparation of traditional Chinese medicine has a better therapeutic effect on visceral pain. Conclusion： The traditional Chinese medicine tra_nsdermal preparation combined with western medicine for the treatment of cancer pain may be a new method for the treatment of cancer pain.

Analgesic effect of oxycodone combined with parecoxib sodium after laparoscopic cholecystectomy and its influence on inflammatory stress response

Objective: To discuss the analgesic effect of oxycodone combined with parecoxib sodium after laparoscopic cholecystectomy and its influence on inflammatory stress response. Methods:A total of 260 patients with chronic cholecystitis who accepted laparoscopic cholecystectomy in this hospital between December 2016 and May 2017 were divided into control group (n=130) and oxycodone group (n=130) by random number table method. Control group received morphine combined with parecoxib sodium analgesia after operation, and oxycodone group received oxycodone combined with parecoxib sodium analgesia after operation. The differences in serum levels of pain mediators, inflammatory mediators and stress hormones were compared between the two groups immediately after operation (T0), 12 h after operation (T1) and 24 h after operation (T3). Results: At T0, there was no statistically significant difference in serum levels of pain mediators, inflammatory mediators and stress hormones between the two groups. At T1 and T2, serum pain mediators PGE2, NPY, SP and NGF levels of oxycodone group were lower than those of control group whereas β-EP levels were higher than those of control group;serum inflammatory mediators CRP, IL-1β, IL-6 and TNF-αlevels were lower than those of control group;serum stress hormones Cor, NE and INS levels were lower than those of control group. Conclusion: Oxycodone combined with parecoxib sodium analgesia after laparoscopic cholecystectomy can effectively relieve the pain perception and inhibit the systemic inflammatory response and stress response.

Effect of oxycodone hydrochloride injection preemptive analgesia on serum inflammatory factors, neurotransmitter index and immune function in patients with laparoscopic cholecystectomy

Objective:To investigate the effect of oxycodone hydrochloride injection preemptive analgesia on serum inflammatory factors, neurotransmitter index and immune function in patients with laparoscopic cholecystectomy.Methods: According to random data table, 113 patients undergoing laparoscopic cholecystectomy were divided into control group (n=57) and observation group (n=56), patients in the control group were treated with sufentanil citrate injection analgesia, and the observation group patients were given oxycodone hydrochloride injection analgesia, level of serum inflammatory factors [tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6)], neurotransmitter index [5-hydroxy tryptamine (5-HT), P substance] and immune function index [CD4+, CD8+, CD4+/CD8+] of two groups between preoperative and postoperative 1d were compared.Results: There were no significant difference in level of TNF-α, IL-6, 5-HT, P substance, CD4+, CD8+ and CD4+/CD8+ between the two groups preoperative. Compared with the level of the same group preoperative, at postoperative 1 d level of TNF-α, IL-6, 5-HT, P substance, CD8+were significantly increased, moreover level in the observation group were significantly lower compared with the control group, the difference was statistically significant;Postoperative 1 d, level of CD4+, CD4+/CD8+ in the two groups were significantly lower than the preoperative level within the group, and the observation group was significantly higher than the control group.Conclusion: Oxycodone hydrochloride injection preemptive analgesia in laparoscopic cholecystectomy can effectively reduce serum inflammatory factors and neurotransmitter index release, improve immune function, has an important clinical value.

Acetaminophen overdose-induced acute liver injury can be alleviated by static magnetic field

Acetaminophen(APAP),the most frequently used mild analgesic and antipyretic drug worldwide,is implicated in causing 46%of all acute liver failures in the USA and between 40%and 70%in Europe.The predominant pharmacological intervention approved for mitigating such overdose is the antioxidant N-acetylcysteine(NAC);however,its efficacy is limited in cases of advanced liver injury or when administered at a late stage.In the current study,we discovered that treatment with a moderate intensity static magnetic field(SMF)notably reduced the mortality rate in mice subjected to high-dose APAP from 40%to 0%,proving effective at both the initial liver injury stage and the subsequent recovery stage.During the early phase of liver injury,SMF markedly reduced APAPinduced oxidative stress,free radicals,and liver damage,resulting in a reduction in multiple oxidative stress markers and an increase in the antioxidant glutathione(GSH).During the later stage of liver recovery,application of vertically downward SMF increased DNA synthesis and hepatocyte proliferation.Moreover,the combination of NAC and SMF significantly mitigated liver damage induced by high-dose APAP and increased liver recovery,even 24 h post overdose,when the effectiveness of NAC alone substantially declines.Overall,this study provides a noninvasive non-pharmaceutical tool that offers dual benefits in the injury and repair stages following APAP overdose.Of note,this tool can work as an alternative to or in combination with NAC to prevent or minimize liver damage induced by APAP,and potentially other toxic overdoses.

莪术二酮对APAP导致急性肝损伤的保护作用

目的 探讨莪术二酮(curdione,CUR)对对乙酰氨基酚(acetaminophen,APAP)诱导急性肝损伤的保护作用。方法 将6~8周龄的C57BL/6雄性小鼠随机分为对照组(CON)、APAP造模组(APAP)、CUR低剂量给药组(25 mg/kg)、CUR高剂量给药组(50 mg/kg),每组6只。连续3 d灌胃给药后,除CON组外,每组小鼠腹腔注射APAP(400 mg/kg)建立肝损伤模型。APAP注射6 h后,取小鼠血清和肝脏,检测血清中天门冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)水平;使用苏木精-伊红对肝组织切片染色,评估肝脏损伤情况;测定肝脏组织中丙二醛(MDA)、超氧化物歧化酶(SOD)的表达水平;应用Western blot检测肝脏组织中凋亡因子(Bax、Bcl-2、Cleaved Caspase 3)及氧化应激因子(Nrf2、HO-1和NQO-1)蛋白表达水平;通过TUNEL染色检测肝细胞凋亡情况。结果 与造模组比较,给药组小鼠血清中AST、ALT水平显著降低(P<0.05或P<0.01);肝组织苏木精-伊红染色显示,CUR可显著缓解小鼠肝脏组织形态结构改变;给药组小鼠肝组织中MDA含量显著降低,SOD活性显著升高(P<0.05或P<0.01);Western blot结果显示,CUR能显著提高氧化应激因子的蛋白表达,凋亡因子Bax和Cleaved Caspase 3蛋白表达显著下调,Bcl-2蛋白表达显著上调(P<0.05或P<0.01);TUNEL染色结果显示,CUR能显著减少APAP诱导的肝细胞死亡。结论 CUR可以显著改善APAP诱导的急性肝损伤。

Chrysanthemum extract alleviates acetaminophen-induced liver injury by inhibiting oxidative stress via AMPK pathway in rats

OBJECTIVE Acetaminophen(APAP),also known as paracetamol,is a commonly used antipyretic,anal⁃gesic and anti-inflammatory drug.However,during the use of APAP for more than half a century,people have not only used APAP to fight diseases but have also suffered the adverse effects brought about by APAP for more than half a cen⁃tury.The most serious adverse reaction to APAP is hepatotoxicity caused by overdose or long-term use.In Chinese tra⁃ditional medicine,chrysanthemums have the functions of dispelling wind,dissipating heat,clearing the liver and improv⁃ing eyesight.Although the chrysanthemum variety named Bianliang ziyu from Kaifeng is not a medicinal variety,it has good value for medicine and food.The aim of this study was to investigate the protective effect of Bianliang Ziyu extract(BZE)on APAP-damaged rats and the potential molecular mechanism.METHODS Male Sprague-Dawley rats(200-220 g)were intragastrically administered BZE(110,220 and 440 mg·kg^-1)for 8 d.On the ninth day,APAP(800 mg·kg^-1)was administered intragastrically to the rats 0.5 h after BZE administration to induced drug-induced liver injury.The serum and liver samples were collected after 24 h.The levels of alanine aminotransferase(ALT),aspartic aminotransferase(AST),reactive oxygen species(ROS),malondialdehyde(MDA),superoxide dismutase(SOD)and glutathione(GSH)in serum and liver tissue of rats were detected by kit method.HE staining was used to observe the histopathological changes in the liver of rat.The effects of BZE on the expression of the oxidative stress related proteins and the mitochondrial biosyn⁃thesis related proteins were detected by Western blot.RESULTS The results showed that BZE significantly reduced the levels of ALT,AST,MDA and ROS and increased the levels of GSH and SOD caused by APAP.Moreover,BZE increased phosphorylation of AMP-activated protein kinase(AMPK)and glycogen synthase kinase 3β(GSK3β),promoted the nuclear translocation of nuclear factor-erythroid 2-related factor 2(Nrf2).BZE also upregulated the expression of mitochondrial biosynthesis related proteins such as peroxisome proliferator-activated receptorγ(PPAR-γ),peroxisome proliferator-activated receptorγcoactivator-1α(PGC-1α),mitochondrial transcription factor(TFAM)and nuclear respira⁃tory factor 1(NRF1).CONCLUSION BZE alleviates APAP-induced liver injury in rats by inhibiting oxidative stress via GSK3β-Nrf2 signaling and the mitochondrial biosynthesis pathway mediated by AMPK.

Effects of a bioartificial liver support system on acetaminophen induced acute liver failure canines

AIM To evaluate the safety and efficacy of the bioartificial liver support system in canines with acute liver failure (ALF). METHODS Nine canines with acute liver failure by acetaminophen induced received TECA Ⅰ bioartificial liver support system (BALSS) from Hong Kong TECA LTD Co. Blood was perfused through a hollow fiber tube containing (1 2)×10 10 the porcine hepatocytes. In contrast, another 10 canines with acute liver failure by Acetaminophen received drugs. Each treatment lasted 6 hours. RESULTS BALSS treatment resulted in beneficial effects for acetaminophen induced ALF canines with survival and with the recovery of the liver functions and tissues, and plasma ammonia decreased from 135 9μmol/L ± 17 5μmol/L to 65 7μmol/L ± 22 0μmol/L , 32 5μmol/L ± 8 8μmol/L , GPT from 97 8U/L ± 8 7U/L to 64 8U/L ± 11 9U/L , 19 0U/L ± 6 3U/L , GOT from 103 0U/L ± 16 7U/L to 75 7U/L ± 19 6U/L , 26 5U/L ± 5 0U/L , and AKP from 158 3U/L ± 12 1U/L to 114 5U/L ± 19 8U/L , 43 8U/L ± 5 6U/L during and after the treatment. In contrast, 10 ALF canines in both the drug and control groups died 1 or 2 days after treatment. CONCLUSION TECA 1 artificial liver support system is safe and efficacious for canines with acute liver failure.

Liuweiwuling tablets attenuate acetaminophen-induced acute liver injury and promote liver regeneration in mice

AIM: To explore the mechanism of protection against acetaminophen-induced acute liver injury by Liuweiwuling tablets.METHODS: Intraperitoneal injections of acetaminophen(250 mg/kg) were used to induce acute liver injury in male C57BL/6 mice.A total of 24 healthy mice were randomly assigned to two groups: an acute liver injury group(control group) and a Liuweiwuling tablet group.Mice were given Liuweiwuling tablets or a vehicle(PBS) orally prior to the administration of acetaminophen.Serum alanine aminotransferase(ALT) and aspartate aminotransaminase(AST) levels were measured at different time points within one week,and pathological examinations of liver tissues were performed 36 h after induction of acute liver injury.Serum inflammatory cytokines,such as high mobility group box protein B1(HMGB1),tumor necrosis factor(TNF)-α and interleukin IL-1b,were detected using an ELISA method according to the manufacturer's instructions.Hepatic morphological changes at 36 h were assessed by hematoxylin and eosin staining.Expression of proliferating cell nuclear antigen(PCNA) in liver tissue was determined by Western blot analysis.The m RNA levels of hepatocyte proliferation markers(PCNA,Cyclin D1 and p21) were detected by real-time quantitative reverse transcription-polymerase chain reaction.RESULTS: The levels of ALT/AST in the Liuweiwuling tablet group were decreased significantly at 6,12 and 24 h compared to that of the control group(654.38 ± 120.87 vs 1566.17 ± 421.64,1154.18 ± 477.72 vs 4654.84 ± 913.71 and 935.13 ± 252.34 vs 4553.75 ± 727.37,P < 0.01).Serum HMGB1 levels at 6 and 12 h for the Liuweiwuling tablet group were significantly lower than those of the control group(23.49 ± 3.89 vs58.6 ± 3.65,61.62 ± 13.07 vs 27.32 ± 5.97,P < 0.01).Furthermore,serum TNF-α and IL-1b levels at 12 h in the Liuweiwuling tablet group were also significantly lower than those of the control group(299.35 ± 50.61 vs 439.03 ± 63.59,57.42 ± 12.98 vs 160.07 ± 49.87,P < 0.01).Centrilobular necrosis was evident in liver tissue of mice with acetaminophen-induced acute liver injury,but was almost abolished in the Liuweiwuling tablet group.The expression levels of PCNA and Cyclin D1 were up-regulated in liver tissue in the Liuweiwuling tablet group(321.08 ± 32.87 vs 157.91 ± 21.52,196.37 ± 25.39 vs 68.72 ± 11.27,P < 0.01); however,expression of p21 in liver tissue was downregulated compared to that of the control group(40.26 ± 9.97 vs 138.24 ± 13.66,P < 0.01).CONCLUSION: Liuweiwuling tablets can attenuate acute liver injury by decreasing inflammatory cytokine(HMGB1,TNF-α and IL-1b) levels and promoting liver regeneration.

Protective effects of Phyllanthus acidus(L.) Skeels leaf extracts on acetaminophen and thioacetamide induced hepatic injuries in Wistar rats

Objective:To investigate and compare the hepatoprotective effects of crude ethanolic and aqueous extracts of Phyllanthus acidus（L.） Skeels（P.acidus） leaves on acetaminophen（APAP） and thioacetamide（TAA） induced liver toxicity in wistar rats.Silymarin was the reference hepatoprotective agent.Methods:In two different sets of experiments,the P.acidus extracts （200 and 400 mg/kg,body weight） and silymarin（100 mg/kg,body weight） were given orally for 7 days and a single dose of APAP（2 g/kg,per oral） or TAA（100 mg/kg,subcutaneous） were given to rats.The level of serum aspartate transaminase（AST）,alanine transaminase（ALT）,alkaline phosphatase（ALP）,total bilirubin and total protein were monitored to assess hepatotoxicity and hepatoprotection.Results:APAP or TAA administration caused severe hepatic damage in rats as evident from significant rise in serum AST,ALT,ALP,total bilirubin and concurrent depletion in total serum protein.The P.acidus extracts and silymarin prevented the toxic effects of APAP or TAA on the above serum parameters indicating the hepatoprotective action.The aqueous extract was found to be more potent than the corresponding ethanolic extract against both toxicants.The phenolic and flavonoid content（175.02±4.35 and 74.68±1.28,respectively） and 2,2-diphenyl-1- picrylhydrazil（DPPH）[IC50=（33.2±0.31）μg/mL]scavenging potential was found maximum with aqueous extract as compared to ethanolic extract.Conclusions:The results of present study suggests that the aqueous extract of P.acidus leaves has significant hepatoprotective activity on APAP and TAA induced hepatotoxicity,which might be associate with its high phenolic and flavonoid content and antioxidant properties.

Curcumin protects against acetaminophen-induced apoptosis in hepatic injury

AIM:To explore the effects of curcumin(CMN)on hepatic injury induced by acetaminophen(APAP)in vivo.METHODS:Male mice were randomly divided into three groups:groupⅠ(control)mice received the equivalent volumes of phosphate-buffered saline(PBS)intraperitoneally(ip);GroupⅡ[APAP+carboxymethylcellulose(CMC)]mice received 1%CMC(vehicle)2h before APAP injection;GroupⅢ(APAP+CMN)mice received curcumin(10 or 20 mg/kg,ip)2 h before before or after APAP challenge.In GroupsⅡandⅢ,APAP was dissolved in pyrogen-free PBS and injected at a single dose of 300 mg/kg.CMN was dissolved in 1%CMC.Mice were sacrificed 16 h after the APAP injection to determine alanine aminotransferase(ALT)levels in serum and malondialdehyde(MDA)accumulation,superoxide dismutase(SOD)activity and hepatocyte apoptosis in liver tissues.RESULTS:Both pre-and post-treatment with curcumin resulted in a significant decrease in serum ALT compared with APAP treatment group(10 mg/kg:801.46±661.34 U/L;20 mg/kg:99.68±86.48 U/L vs 5406.80±1785.75 U/L,P<0.001,respectively).The incidence of liver necrosis was significantly lowered in CMN treated animals.MDA contents were significantly reduced in 20 mg/kg CMN pretreatment group,but increased in APAP treated group(10.96±0.87 nmol/mg protein vs 16.03±2.58 nmol/mg protein,P<0.05).The decrease of SOD activity in APAP treatment group and the increase of SOD in 20 mg/kg CMN pretreatment group were also detected(24.54±4.95 U/mg protein vs 50.21±1.93 U/mg protein,P<0.05).Furthermore,CMN treatment efficiently protected against APAPinduced apoptosis via increasing Bcl-2/Bax ratio.CONCLUSION:CMN has significant therapeutic potential in both APAP-induced hepatotoxicity and other types of liver diseases.