CN104388536A - Hepatocyte-based medicine hepatotoxicity evaluation method - Google Patents
Hepatocyte-based medicine hepatotoxicity evaluation method Download PDFInfo
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Abstract
The invention discloses a hepatocyte-based medicine hepatotoxicity evaluation method. The method comprises the following steps: 1) culturing original liver-inhibiting cells; 2) dividing the original liver-inhibiting cells into three groups which are respectively a group in which medicines to be tested are added, a mixture group in which medicines to be tested and cholic acid compounds are added, and a negative control group; 3) respectively detecting the content of ALT/AST/LDH and urea in a culture medium, and computing the amount of the generated urea through the content of the urea; 4) comparing the content of ALT/AST/LDH, the content of urea or the amount of the generated urea in the three groups of the culture mediums, and evaluating the medicine hepatotoxicity to be tested. The method uses the dual-layer collagen three-dimensional-culture technology to simulate the environment in a body, induces the hepatocyte to form a hepatic plate sample structure and a bile canaliculus network, and in the culture process, maintains the biological function of the hepatocyte; at the same time, cholic acid-like compounds are added to simulate the environment of the hepatocyte in the body, so that the hepatotoxicity screening is performed on the medicines.
Description
Technical field
The invention belongs to cell cultures and biochemistry index test field, relate to a kind of use in medicament-induced hepatotoxicity appraisal procedure, particularly relate to a kind of based on hepatocellular use in medicament-induced hepatotoxicity appraisal procedure
Background technology
Liver is human body vitals, is responsible for the different physiological roles such as metabolism, removing toxic substances, blood coagulation, immunity.And in drug use process, medicine and/or its meta-bolites may cause hepar damnification, be called drug induced hepatic injury (DILI).DILI is that 50 years medicines are because the modal reason in city is removed in security in the past, even if Wei Che city, the liver toxicity of medicine also can limit the clinical application of a lot of medicine.Because most medicine causes the probability of serious DILI very low (≤1/10000), so usually cannot be identified at clinical trial, become a difficult problem for puzzlement drugmaker.
Whether usual researchist mainly utilizes the index such as LDH seepage, albumin secretion, urea generation the liver injury of cell model or animal model can be caused to screen to medicine.But, species difference likely judges to have an impact to liver toxicity, in human body, cause the medicine of serious DILI in animal experiment, not show clear and definite liver toxicity, generally also non-show dose related toxicity, much cause serious hepatotoxic medicine but nontoxicity on animal model.There is obvious Cytotoxic drug candidate namely can be excluded in early days in research and development simultaneously, cause the medicine of serious DILI often directly can't cause cytotoxicity in clinical or listing stage.Can be given special attention the medicine that AST or ALT can be caused to exceed normal range value more than three times during clinical experiment, because they may cause serious liver injury, if but direct testing drug is to the toxicity of primary cultured hepatocyte, often ignoring after those are taken medicine causes cholate in liver, bilirubin concentration to raise, thus causes the medicine of hepar damnification.And the clinical medicine causing DILI usually tolerance is good, but due to private medical service different, in only a few crowd, just may can produce liver injury.As can be seen here, existing use in medicament-induced hepatotoxicity detection method can not reflect the liver damage score of medicine reality completely, this area be badly in need of a kind of can the hepatotoxic method of accurate evaluation medicine.
Summary of the invention
The object of the invention is to propose one based on hepatocellular use in medicament-induced hepatotoxicity appraisal procedure, it utilizes double-layer collagen dimensional culture technology to carry out simulated in vivo environment, inducing hepatocyte forms liver plate spline structure and bile canaliculus network, hepatocellular biological function is maintained in culturing process, as platform, liver toxicity screening is carried out to medicine.
For reaching this object, the present invention by the following technical solutions:
A kind of based on hepatocellular use in medicament-induced hepatotoxicity appraisal procedure, it comprises the following steps:
1) primary hepatocyte is cultivated;
2) described primary hepatocyte is divided into three groups, is respectively the group adding medicine to be measured, the group adding medicine to be measured and Cholic acids compound and negative control group;
3) detect alanine aminotransferase (ALT)/aspartate amino transferase (AST)/serum lactic dehydrogenase (LDH) content in substratum and urea content respectively, and calculate urea growing amount by urea content;
4) the ALT/AST/LDH content in three groups of substratum or urea growing amount are compared, assess use in medicament-induced hepatotoxicity to be measured.
First method of the present invention carries out hepatocyte model structure, and liver model used can comprise the hepatocyte model built with various training method, as suspension culture, adherent culture and dimensional culture.But dimensional culture mode can build the little tube model of liver and gall, more advantageously can assess use in medicament-induced hepatotoxicity.
Therefore, in step 1) in, described cultivation is double-layer collagen dimensional culture, and its inducing hepatocyte forms liver plate spline structure and bile canaliculus network.Described double-layer collagen dimensional culture technology is the substratum added on collagen containing 5%-10% foetal calf serum, then adds not containing serum, substratum containing matrigel and/or collagen, forms sandwich structure and cultivates.Preferably, 2-6h is cultivated after adding the substratum containing 5%-10% foetal calf serum.
Described primary hepatocyte be from human liver tissue be separated cell or animal liver cell, described animal comprises rat, mouse, dog, pig, cat, ox, sheep, chicken, horse, goose, duck and primates, and described primates comprises ape, tree shrew and monkey; Preferably, the preparation of cell prepares through two step collagen perfusion method digestion.
Preferably, consider the otherness of species, select human liver cell to carry out hepatocyte model structure as primary stem cell, the human liver cell dimensional culture model with bile duct network can be formed especially.In addition, consider the individual difference of people, should adopt >=human liver cell of the different donors of 3 to carry out the liver toxicity assessment under same experimental conditions, reduce with this error that individual difference produces.
Before described drug effect, the cell cultures time is 1-4 days, and cultivate the 1st day to the 4th day, three-dimensional structure is perfect gradually, to form liver plate spline structure and bile canaliculus network.
In step 2) in, after the liver cell cultivated is adherent, medicine to be measured, Cholic acids compound or reference substance can be added.Described Cholic acids compound is glycochenodeoxycholate GCDCA, Chenodiol CDCA, glycodesoxycholic acid GDCA, Septochol DCA, the free acid of glycocholic acid GCA or its inorganic salt; Described inorganic salt are preferably sodium salt; Preferably, after adding each material, incubation time is 1-3 days.
Existing method mostly direct testing drug, to the toxicity of primary cultured hepatocyte, is often ignored after those are taken medicine and is caused cholate in liver, bilirubin concentration to raise, thus cause the medicine of hepar damnification.The present invention detects in culture system and adds Cholic acids compound, environment residing for liver cell in energy analogue body.For determining the concentration of Cholic acids compound suitable in system, the present invention adopts the Cholic acids compound effects of a series of different concns in human liver cell, with the IC of urea growing amount for the various Cholic acids compound of index calculate
50value:
According to IC
50under the downward of the concentration of Cholic acids compound is made this concentration by value, described Cholic acids compound can not cause hepatocellular ALT/AST/LDH and urea growing amount to change, in order to avoid affect detected result.Cholic acids compound in the following concentration range of empirical tests can not cause obvious ALT/AST/LDH to discharge the change of increase and urea growing amount.Concrete preferred concentration range is as follows:
The add-on of described medicine to be measured is based on the character of medicine itself and conventional administration amount thereof, and those skilled in the art can select according to the conventional administration concentration of known drug.Described negative control group adds the blank solvent not containing medicine to be measured and/or Cholic acids compound, and itself and all the other quantity of solvent of two groups is identical, and preferred solvent is DMSO.
In step 3) in, described detection ALT/AST/LDH content, for carry out substratum replacing before detection, detects the ALT/AST/LDH content in the substratum be removed; Preferably, automatic clinical chemistry analyzer is used to detect ALT/AST/LDH content;
Preferably, described detection urea content comprises: carry out substratum replacing before detection, detects the urea content in the substratum that is removed, or described step 2) cultivation after add the urea content detected after blank cultures cultivates 1 hour in substratum.The two kinds of urea content values recorded respectively according to these two kinds of methods all can be used for calculating urea growing amount, as the judge index of the method for the invention.
Preferably, diacetyl monoxime method is used to detect urea content;
The method of calculation of described urea growing amount are:
Urea growing amount=(urea content × described culture volume)/incubation time.
After obtaining ALT/AST/LDH content and urea growing amount, these two kinds of indexs are compared.
In step 4) in, describedly to compare for the group adding medicine to be measured, the group that adds medicine to be measured and Cholic acids compound are compared with the urea growing amount of negative control group and ALT/AST/LDH content.
When the group adding medicine to be measured, the ALT/AST/LDH content that adds the group of medicine to be measured and Cholic acids compound obviously raise and urea growing amount reduces, show that this medicine is simple cytotoxicity;
When the ALT/AST/LDH content of the group adding separately medicine to be measured compares negative control group without considerable change with urea growing amount, but adding the ALT/AST/LDH content showed increased of the group of medicine to be measured and Cholic acids compound and/or urea growing amount when obviously reducing, showing that this medicine may be by causing cholestasis to cause liver toxicity.
Present method utilizes primary cultured hepatocyte as platform, compares adding medicine to be measured with the ALT/AST/LDH added in the substratum of medicine to be measured and Cholic acids compound and urea growing amount, and release and the urea of investigating ALT/AST/LDH generate.Wherein the release of ALT/AST/LDH embodies liver injury biochemical indicator, and urea generates embodiment liver cell function of detoxification.This method can not only evaluate use in medicament-induced hepatotoxicity, can also distinguish medicine to be measured and cause hepatotoxic approach.
The present invention also aims to provide the application of described method in detection of drugs liver toxicity.
The method of the invention utilizes double-layer collagen dimensional culture technology to carry out simulated in vivo environment, inducing hepatocyte forms liver plate spline structure and bile canaliculus network, hepatocellular biological function is maintained in culturing process, add Cholic acids compound glycochenodeoxycholate GCDCA simultaneously, Chenodiol CDCA, Septochol DCA, glycodesoxycholic acid GDCA, the free acid of glycocholic acid GCA or the mixture of its salt are with environment residing for liver cell in analogue body, thus liver toxicity screening is carried out to medicine, the drug candidate that may cause severe liver injury can be found in early days, save research and development time and cost.It is consistent with the liver toxicity that this medicine is reported that the method for the invention surveys result to the liver toxicity of known drug, prove that present method is accurately available, method construct outer platform of the present invention simultaneously, have cost less, the advantage such as the time is short, end user source liver cell, avoids species difference to affect liver toxicity and judges.
Accompanying drawing explanation
Fig. 1 is the double-layer collagen dimensional culture primary human liver cell Photomicrograph of 24,48,72,96 hours.
Fig. 2 utilizes the fluorogenic substrate through bile duct excretion to form cholangiolar Photomicrograph to show the primary human liver cell of double-layer collagen dimensional culture after 96 hours.
Fig. 3 after 1 day, feeds ciclosporin A and ciclosporin A and the effect of the Cholic acids compound human liver cell form Photomicrograph after 24 hours in dimensional culture.
Fig. 4 after 4 days, feeds ciclosporin A and ciclosporin A and the effect of the Cholic acids compound human liver cell form Photomicrograph after 24 hours in dimensional culture.
Embodiment
Technical scheme of the present invention is further illustrated by embodiment below in conjunction with accompanying drawing.
Embodiment 1 cultivates primary human liver cell
Double-layer collagen dimensional culture:
Use two step collagen perfusion methods that the liver organization block isolated perfusion of people is digested to liver cell.After calculating cell number and motility rate with blood counting chamber and Trypan Blue, being diluted to the substratum containing 5%-10% foetal calf serum is covered with in the Tissue Culture Plate of collagen, cultivate and to change into after 2-6h containing serum, substratum containing matrigel and/or collagen, form double-layer collagen three-dimensional structure and cultivate.It cultivates 24,48,72, photo after 96h is shown in Fig. 1.Visible from 24h, liver cell forms liver plate spline structure and bile canaliculus network gradually, extends the bile canaliculus network closer to human body in time, (forms more stable bile canaliculus structure after double-layer collagen dimensional culture primary hepatocyte 96h) as shown in Figure 2.
The determination of embodiment 2 Cholic acids compound concentration
For determining the concentration of Cholic acids compound suitable in system, the present invention adopts the Cholic acids compound effects of a series of different concns in human liver cell, with the IC of urea growing amount for the various Cholic acids chemical combination of index calculate
50value.
According to IC
50under the downward of Cholic acids compound concentration is made this concentration by value, described Cholic acids compound can not cause hepatocellular ALT/AST/LDH and urea generation to change, in order to avoid affect detected result.The mixture of the Cholic acids compound in the following concentration range of empirical tests all can not cause obvious ALT/AST/LDH to discharge and ureagenetic change.Concrete concentration range is:
Embodiment 3 is based on hepatocellular use in medicament-induced hepatotoxicity appraisal procedure
Use the liver cell that embodiment 1 is cultivated, the Cholic acids compound concentration determined according to embodiment 2 carries out following steps:
1) with the kind plate density of every square centimeter of 150,000 cells inoculation primary hepatocyte, and in 37 DEG C of incubators, the dimensional culture of double-layer collagen is carried out;
2) cell is divided into three groups, adds medicine to be measured, medicine to be measured and Cholic acids compound respectively and without pharmaceutically-active negative control;
3) the urea growing amount of (as in 1 hour) in the liver cell unit time after ALT/AST/LDH in substratum and urea content or drug effect is detected;
4) ALT/AST/LDH in three groups of substratum and urea content value or urea growing amount are compared, assess use in medicament-induced hepatotoxicity to be measured.
The human liver cell of the various medicine of embodiment 4 detects
According to embodiment 3, liver toxicity detection is carried out to the various medicines in form.The results are shown in Table 1.
By human liver cell dimensional culture after 1 day, add separately after medicine and medicine and Cholic acids compound and human liver cell hatch 24h, measure ALT/AST/LDH/ urea content, result is as follows:
Table 1
Foreign literature is reported, cyclosporine A hepatotoxicity sickness rate can reach 20% ~ 40%; Domestic literature reports that its sickness rate is 36.5% ~ 46.8%, and can increase the weight of the liver toxicity of ciclosporin A during cholestasis.So ciclosporin A is always as the positive control of liver toxicity research.For liver toxicity research, conventional transaminase (ALT and AST) and total bilirubin level are evaluated clinically, and In vitro cell experiment commonly uses LDH release and Cell viability is evaluated, the dimensional culture models coupling evaluation index of clinical and common experiment in vitro in the present invention, adopts transaminase (ALT and AST), LDH release and urea to generate comprehensive evaluation medicine to hepatocellular toxic action.
The result of table 1 also demonstrates the hepatotoxicity of ciclosporin A.And when alone ciclosporin A does not directly produce obvious liver toxicity, (ALT/AST/LDH raises less than 2 times, urea generates minimizing and is no more than 40%), conbined usage medicine and Cholic acids compound are relative to alone medicine, ALT/AST/LDH increases to over 3 times, and urea generates and reduces by more than 40%.
And the change contrasting the microscopic morphology of cell also can find out the liver toxicity of ciclosporin.In dimensional culture after 1 day or 4 days, the human liver cell form Photomicrograph of mixture effect after 24 hours feeding ciclosporin A and ciclosporin A and Cholic acids compound is distinguished as shown in Figure 3 and Figure 4.Cell microscopic morphology in comparison diagram 3 and Fig. 4, can obtain drawing a conclusion: 1) contrast blank group (without medicine, without Cholic acids compound) and only add Cholic acids compound group, in the Cholic acids compound concentration range that the present invention limits, Cholic acids compound on intracellular is without obvious damaging action; 2) contrast adds ciclosporin A group and the blank group of 20uM, and visible ciclosporin A has certain damaging action (on figure display section cell occur that structure is imperfect, swelling) to liver cell; 3) contrast only adds the ciclosporin A group of 20uM and adds ciclosporin A and Cholic acids compound group, visible the latter can increase the weight of hepatocellular damage (figure shows most cells structure is imperfect, fragmentation even dead), ciclosporin cause alluvial in cell this is because can suppress cholic acid to be arranged outward, thus increase the weight of hepatocellular damage, also consistent to the liver toxicity result of study of ciclosporin with document.Can directly cause the paracetamol of liver injury then because concentration does not show liver toxicity far below the Plasma Concentration (>6mM) during clinical administration, this illustrates that the liver toxicity of this medicine is concentration dependent, consistent with bibliographical information and clinical application.
The rat hepatocytes of the various medicine of embodiment 5 detects
Take in situ perfusion to digest, the liver organization of rat is digested to liver cell.Rest part, according to embodiment 3, carries out liver toxicity detection to the various medicines in form.The results are shown in Table 2.
After rat hepatocytes being cultivated 1 day, add separately after medicine and medicine and Cholic acids compound and rat hepatocytes hatch 24h, measure ALT/AST/LDH/ urea content, result is as follows:
Table 2
The result of rat hepatocytes model and the result of human liver cell similar, namely when alone medicine does not directly produce obvious liver toxicity, (ALT/AST/LDH raises less than 2 times, urea generates minimizing and is no more than 40%), conbined usage medicine and Cholic acids Compound Phase are for alone medicine, ALT/AST/LDH increases to over 3 times, and urea generates and reduces by more than 40%.Associative list 1 and table 2, this model successfully filters out and clinically may cause cholestasis thus the ciclosporin A of liver injury and chlorpromazine, consistent with bibliographical information.
The method of the invention utilizes double-layer collagen dimensional culture technology to carry out simulated in vivo environment, inducing hepatocyte forms liver plate spline structure and bile canaliculus network, hepatocellular biological function is maintained in culturing process, add Cholic acids compound glycochenodeoxycholate GCDCA simultaneously, Chenodiol CDCA, Septochol DCA, glycodesoxycholic acid GDCA, the free acid of glycocholic acid GCA or the mixture of its salt are with environment residing for liver cell in analogue body, thus liver toxicity screening is carried out to medicine, the drug candidate that may cause severe liver injury can be found in early days, thus greatly save research and development time and cost.The liver toxicity detected result of the method for the invention to known drug is consistent with the liver toxicity that this medicine is reported, prove that present method is accurately available, method of the present invention is a kind of external platform simultaneously, have cost less, the advantage such as the time is short, and end user source liver cell, species difference can be avoided to affect liver toxicity and to judge.
Applicant states, the present invention illustrates method detailed of the present invention by above-described embodiment, but the present invention is not limited to above-mentioned method detailed, does not namely mean that the present invention must rely on above-mentioned method detailed and could implement.Person of ordinary skill in the field should understand, any improvement in the present invention, to equivalence replacement and the interpolation of ancillary component, the concrete way choice etc. of each composition of product of the present invention, all drops within protection scope of the present invention and open scope.
Claims (9)
1., based on a hepatocellular use in medicament-induced hepatotoxicity appraisal procedure, it is characterized in that, said method comprising the steps of:
1) primary hepatocyte is cultivated;
2) described primary hepatocyte is divided into three groups, is respectively the group adding medicine to be measured, the group adding medicine to be measured and Cholic acids compound and negative control group;
3) detect the ALT/AST/LDH content in substratum and urea content respectively, and calculate urea growing amount by urea content;
4) the ALT/AST/LDH content in three groups of substratum or urea growing amount are compared, assess use in medicament-induced hepatotoxicity to be measured.
2. method according to claim 1, is characterized in that, in step 1) in, described cultivation is double-layer collagen dimensional culture, and its inducing hepatocyte forms liver plate spline structure and bile canaliculus network;
Preferably, described primary hepatocyte be from human liver tissue be separated cell or animal liver cell, described animal comprises rat, mouse, dog, pig, cat, ox, sheep, chicken, horse, goose, duck and primates, and described primates comprises ape, tree shrew and monkey;
Preferably, the time of described cultivation primary hepatocyte is 1-4 days.
3. method according to claim 2, it is characterized in that, described double-layer collagen dimensional culture technology is that the substratum added containing 5%-10% foetal calf serum makes liver cell adherent on collagen, change into more not containing serum, substratum containing matrigel and/or collagen, cultivate to form sandwich structure.
4. method according to claim 3, is characterized in that, adds and cultivates 2-6h containing after 5%-10% foetal calf serum substratum.
5. method according to claim 1, it is characterized in that, in step 2) in, described Cholic acids compound is free acid or its inorganic salt of glycochenodeoxycholate GCDCA, Chenodiol CDCA, Septochol DCA, glycocholic acid GCA and glycodesoxycholic acid GDCA;
Preferably, described inorganic salt are sodium salt;
Preferably, after adding each material, incubation time is 1-3 days.
6. method according to claim 5, is characterized in that, the concentration of described Cholic acids compound in system is:
7. method according to claim 1, is characterized in that, in step 3) in, described detection ALT/AST/LDH content, for carry out substratum replacing before detection, detects the ALT/AST/LDH content in the substratum be removed;
Preferably, automatic clinical chemistry analyzer is used to detect ALT/AST/LDH content;
Preferably, described detection urea content comprises: carry out substratum replacing before detection, detects the urea content in the substratum that is removed, or described step 2) cultivation after add the urea content detected after blank cultures cultivates 1 hour in substratum;
Preferably, diacetyl monoxime method is used to detect urea content.
8. method according to claim 1, it is characterized in that, in step 4) in, describedly to compare for the group adding medicine to be measured, the group that adds medicine to be measured and Cholic acids compound are compared with the ALT/AST/LDH content of negative control group and urea growing amount.
9. the application of the method according to claim 1-8 in detection of drugs liver toxicity.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105255994A (en) * | 2015-11-13 | 2016-01-20 | 浙江大学 | Method for assessing influence of soil antibiotic pollution on human health |
| WO2017085119A1 (en) * | 2015-11-16 | 2017-05-26 | Insphero Ag | Method and assay for the assessment of a cholestatic risk of a compound |
| CN107949635A (en) * | 2015-03-20 | 2018-04-20 | 休雷尔公司 | For characterizing time-based hepatotoxic method |
| CN109337863A (en) * | 2018-10-25 | 2019-02-15 | 南京鼓楼医院 | A kind of external model construction method that predictive compound is viral to fatty liver |
| CN109564210A (en) * | 2016-03-14 | 2019-04-02 | 银丝佛若有限公司 | Method based on 3D tissue cultures assesses injury of mitochondria |
| CN109964129A (en) * | 2016-09-16 | 2019-07-02 | 库里斯特转运体解决方案有限责任公司 | Cause the method and system of systemic toxicity or hepatotoxic potential for screening candidate compound |
| CN111979183A (en) * | 2020-08-10 | 2020-11-24 | 创芯国际生物科技(广州)有限公司 | Drug hepatotoxicity evaluation method based on liver organoid model |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012143514A1 (en) * | 2011-04-20 | 2012-10-26 | Asociación Centro De Investigación Cooperativa En Biociencias-Cic Biogune | Method for the diagnosis of liver injury based on a metabolomic profile |
-
2014
- 2014-10-27 CN CN201410581692.3A patent/CN104388536A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012143514A1 (en) * | 2011-04-20 | 2012-10-26 | Asociación Centro De Investigación Cooperativa En Biociencias-Cic Biogune | Method for the diagnosis of liver injury based on a metabolomic profile |
Non-Patent Citations (4)
| Title |
|---|
| SALMAN R. KHETANI 等: "The Use of Micropatterned Co-cultures to Detect Compounds that Cause Drug induced Liver Injury in Humans", 《TOXICOLOGICAL SCIENCES》 * |
| 傅青春 等: "药物性肝损伤的发病机制", 《医学与哲学》 * |
| 王晓今 等: "231例药物性肝损伤临床分析", 《肝脏》 * |
| 高绪聪 等: "药物性肝损伤的生物标志物及其评价的研究进展", 《中国药理学与毒理学杂志》 * |
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| CN107949635A (en) * | 2015-03-20 | 2018-04-20 | 休雷尔公司 | For characterizing time-based hepatotoxic method |
| CN105255994A (en) * | 2015-11-13 | 2016-01-20 | 浙江大学 | Method for assessing influence of soil antibiotic pollution on human health |
| WO2017085119A1 (en) * | 2015-11-16 | 2017-05-26 | Insphero Ag | Method and assay for the assessment of a cholestatic risk of a compound |
| CN109564210A (en) * | 2016-03-14 | 2019-04-02 | 银丝佛若有限公司 | Method based on 3D tissue cultures assesses injury of mitochondria |
| CN109964129A (en) * | 2016-09-16 | 2019-07-02 | 库里斯特转运体解决方案有限责任公司 | Cause the method and system of systemic toxicity or hepatotoxic potential for screening candidate compound |
| CN109964129B (en) * | 2016-09-16 | 2023-04-14 | 库里斯特转运体解决方案有限责任公司 | Methods and systems for screening candidate compounds for potential to cause systemic toxicity or hepatotoxicity |
| CN109337863A (en) * | 2018-10-25 | 2019-02-15 | 南京鼓楼医院 | A kind of external model construction method that predictive compound is viral to fatty liver |
| CN111979183A (en) * | 2020-08-10 | 2020-11-24 | 创芯国际生物科技(广州)有限公司 | Drug hepatotoxicity evaluation method based on liver organoid model |
| CN111979183B (en) * | 2020-08-10 | 2021-05-28 | 创芯国际生物科技(广州)有限公司 | Drug hepatotoxicity evaluation method based on liver organoid model |
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