Application of Linear Regression Model of Gpnmb Gene in Rat Injury Time Estimation

doi:10.12116/j.issn.1004-5619.2020.400907

Abstract

Abstract:

Objective To investigate the effects of injury time, postmortem interval （PMI） and postmortem storage temperature on mRNA expression of glycoprotein non-metastatic melanoma protein B （Gpnmb）, and to establish a linear regression model between Gpnmb mRNA expression and injury time, to provide aimed at providing potential indexes for injury time estimation. Methods Test group SD rats were anesthetized and subjected to blunt contusion and randomly divided into 0 h, 4 h, 8 h, 12 h, 16 h, 20 h and 24 h groups after injury, with 18 rats in each group. After cervical dislocation, 6 rats in each group were collected and stored at 0 ℃, 16 ℃ and 26 ℃, respectively. The muscle tissue samples of quadriceps femoris injury were collected at 0 h, 12 h and 24 h postmortem at the same temperature. The grouping method and treatment method of the rats in the validation group were the same as above. The expression of Gpnmb mRNA in rat skeletal muscle was detected by RT-qPCR. The Pearson correlation coefficient was used to evaluate the correlation between Gpnmb mRNA expression and injury time, PMI, and postmortem storage temperature. SPSS 25.0 software was used to construct a linear regression model, and the validation group data was used for the back-substitution test. Results The expression of Gpnmb mRNA continued to increase with the prolongation of injury time, and the expression level was highly correlated with injury time （P<0.05）, but had little correlation with PMI and postmortem storage temperature （P>0.05）. The linear regression equation between injury time （y） and Gpnmb mRNA relative expression （x） was y=0.611 x+4.489. The back-substitution test proved that the prediction of the model was accurate. Conclusion The expression of Gpnmb mRNA is almost not affected by the PMI and postmortem storage temperature, but is mainly related to the time of injury. Therefore, a linear regression model can be established to infer the time of injury.

Key words: forensic pathology, wound age estimation, postmortem interval, postmortem storage temperature, glycoprotein non-metastatic melanoma protein B （Gpnmb）, rats

CLC Number:

DF795.1

Yan-ru XI, Yuan-xin LIU, Na FENG, Zhen GU, Jun-hong SUN, Jie CAO, Qian-qian JIN, Qiu-xiang DU. Application of Linear Regression Model of Gpnmb Gene in Rat Injury Time Estimation[J]. Journal of Forensic Medicine, 2022, 38(4): 468-472.

Figures/Tables 3

References 24

1	孙俊红，朱细燕，张小红，等. 大鼠骨骼肌损伤修复中FZD4时序性变化及其应用[J].中国法医学杂志，2016，31（5）：444-447. doi：10.13618/j.issn.1001-5728.2016.05.004 .
	SUN J H， ZHU X Y， ZHANG X H， et al. Time-dependent expression of FZD4 in contused skeletal muscle of rat used for wound age estimation[J]. Zhongguo Fayixue Zazhi，2016，31（5）：444-447.
2	DU Q X， LI N， DANG L H， et al. Temporal expression of wound healing-related genes inform wound age estimation in rats after a skeletal muscle contusion： A multivariate statistical model analysis[J]. Int J Legal Med，2020，134（1）：273-282. doi：10.1007/s00414-018-01990-2 .
3	陶丽，马剑龙，陈龙. 早期死亡时间推断研究进展[J].法医学杂志，2016，32（6）：444-447. doi：10.3969/j.issn.1004-5619.2016.06.013 .
	TAO L， MA J L， CHEN L， et al. Research progress on estimation of early postmortem interval[J]. Fayixue Zazhi，2016，32（6）：444-447.
4	刘季，宋贞柱，谢润红，等. 大鼠死后脑组织RNA降解与死亡时间推断的研究[J].中国法医学杂志，2007，22（4）：226-228，232. doi：10.3969/j.issn.1001-5728.2007.04.004 .
	LIU J， SONG Z Z， XIE R H， et al. A pilot study on cerebral RNA degradation and its applicability to estimation of postmortem interval in rats[J]. Zhongguo Fayixue Zazhi，2007，22（4）：226-228，232.
5	MA J L， PAN H， ZENG Y， et al. Exploration of the R code-based mathematical model for PMI estimation using profiling of RNA degradation in rat brain tissue at different temperatures[J]. Forensic Sci Med Pathol，2015，11（4）：530-537. doi：10.1007/s12024-
	015-9703-7.
6	LI C， WANG Q， ZHANG Y， et al. Research progress in the estimation of the postmortem interval by Chinese forensic scholars[J]. Forensic Sci Res，2016，1（1）：3-13. doi：10.1080/20961790.2016.1229377 .
7	ZHOU L， ZHUO H， OUYANG H， et al. Glycoprotein non-metastatic melanoma protein b （Gpnmb） is highly expressed in macrophages of acute injured kidney and promotes M2 macrophages polarization[J]. Cell Immunol，2017，316：53-60. doi：10.1016/j.cellimm. 2017.03.006 .
8	HUANG J J， MA W J， YOKOYAMA S. Expression and immunolocalization of Gpnmb， a glioma-associated glycoprotein， in normal and inflamed central nervous systems of adult rats[J]. Brain Behav，2012，2（2）：85-96. doi：10.1002/brb3.39 .
9	SUN J H， WANG Y Y， ZHANG L， et al. Time-dependent expression of skeletal muscle troponin I mRNA in the contused skeletal muscle of rats： A possible marker for wound age estimation[J]. Int J Legal Med，2010，124（1）：27-33. doi：10.1007/s00414-009-0323-1 .
10	杜秋香，程晓花，朱细燕，等. 双内参基因用于大鼠挫伤肌肉TAB2 mRNA表达量检测[J].中国法医学杂志，2016，31（3）：231-234，237. doi：10.13618/j.issn.1001-5728.2016.03.003 .
	DU Q X， CHENG X H， ZHU X Y， et al. Application of multiple reference genes for TAB2 mRNA expression in rat skeletal muscle[J]. Zhongguo Fayi-xue Zazhi，2016，31（3）：231-234，237.
11	NIU J， AN G， GU Z， et al. Analysis of sensitivity and specificity： Precise recognition of neutrophils during regeneration of contused skeletal muscle in rats[J]. Forensic Sci Res，2020，7（2）：228-237. doi：10.1080/20961790.2020.1713432 .
12	LI N， DU Q， BAI R， et al. Vitality and wound-age estimation in forensic pathology： Review and future prospects[J]. Forensic Sci Res，2020，5（1）：15-24. doi：10.1080/20961790.2018.1445441 .
13	杜秋香，朱细燕，董塔娜，等. 不同功能基因mRNA推断损伤时间的指标同质性[J].法医学杂志，2018，34（5）：487-491. doi：10.12116/j.issn.1004-5619.2018.05.006 .
	DU Q X， ZHU X Y， DONG T N， et al. Homogeneity of different functional mRNA indicators for wound age estimation[J]. Fayixue Zazhi，2018，34（5）：487-491.
14	董塔娜，裴明，李娜，等. 大鼠骨骼肌挫伤后Fzd2表达与损伤时间的关系[J].法医学杂志，2017，33（4）：344-347. doi：10.3969/j.issn.1004-5619.2017.04.002 .
	DONG T N， PEI M， LI N， et al. Relationship between wound age and the expression of Fzd2 in rats skeletal muscle after contusion[J]. Fayixue Zazhi，2017，33（4）：344-347.
15	HENSSGE C， MADEA B. Estimation of the time since death in the early post-mortem period[J]. Forensic Sci Int，2004，144（2/3）：167-175. doi：10.1016/j.forsciint.2004.04.051 .
16	’T HOEN P A， HIRSCH M， DE MEIJER E J， et al. mRNA degradation controls differentiation state-dependent differences in transcript and splice variant abundance[J]. Nucleic Acids Res，2011，39（2）：556-566. doi：10.1093/nar/gkq790 .
17	ZHU Y， WANG L， YIN Y， et al. Systematic analysis of gene expression patterns associated with postmortem interval in human tissues[J]. Sci Rep，2017，7（1）：5435. doi：10.1038/s41598-017-05882-0 .
18	ROSE A A， SIEGEL P M. Emerging therapeutic targets in breast cancer bone metastasis[J]. Future Oncol，2010，6（1）：55-74. doi：10.2217/fon.09.138 .
19	于天水，官大威，赵锐，等. 大鼠骨骼肌损伤后PMN、MNC及FBC百分率与损伤时间关系[J].法医学杂志，2014，30（3）：166-168. doi：10.3969/j.issn.1004-5619.2014.02.04 .
	YU T S， GUAN D W， ZHAO R， et al. Correlation between percentages of PMN， MNC， FBC and wound age after skeletal muscle injury in rats[J]. Fayixue Zazhi，2014，30（3）：166-168.
20	HARALANOVA-ILIEVA B， RAMADORI G， ARM-BRUST T. Expression of osteoactivin in rat and human liver and isolated rat liver cells[J]. J Hepatol，2005，42（4）：565-572. doi：10.1016/j.jhep.2004.12.021 .
21	MURUGESAN V， LIU J， YANG R， et al. Validating glycoprotein non-metastatic melanoma B （gpNMB， osteoactivin）， a new biomarker of Gaucher disease[J]. Blood Cells Mol Dis，2018，68：47-53. doi：10.1016/j.bcmd.2016.12.002 .
22	BAI X， PENG D， LI Z， et al. Postmortem interval （PMI） determination by profiling of HAF mRNA degradation using RT-qPCR[J]. Forensic Sci Int Genet Suppl Ser，2017，6：e182-e183. doi：10.1016/j.fsigss. 2017.09.072 .
23	TAO L， MA J， HAN L， et al. Early postmortem interval estimation based on Cdc25b mRNA in rat cardiac tissue[J]. Leg Med （Tokyo），2018，35：18-24. doi：10.1016/j.legalmed.2018.09.004 .
24	LV Y H， MA J L， PAN H， et al. RNA degradation as described by a mathematical model for postmortem interval determination[J]. J Forensic Leg Med，2016，44：43-52. doi：10.1016/j.jflm.2016.08.015 .

组别	死后0 h	0 ℃		16 ℃		26 ℃
组别	死后0 h	死后12 h	死后24 h	死后12 h	死后24 h	死后12 h	死后24 h
损伤后0 h	1.00±0.25	0.76±0.20	1.50±0.31	2.20±0.50	1.52±0.15	1.87±0.35	0.72±0.15
损伤后4 h	1.97±0.46	2.76±0.68	1.58±0.18	2.21±0.36	1.79±0.42	2.31±0.41	7.02±2.08
损伤后8 h	2.53±0.50	3.51±0.83	4.33±1.16	4.00±0.99	3.64±1.31	5.10±0.97	9.84±2.50
损伤后12 h	9.20±1.23	13.87±4.51	16.75±3.32	7.98±1.83	12.52±2.45	5.52±1.69	12.49±3.47
损伤后16 h	13.71±4.14	16.57±5.46	13.38±1.80	18.33±5.13	15.56±7.90	21.37±8.30	16.64±1.88
损伤后20 h	15.48±6.27	19.75±2.31	25.71±9.46	23.23±5.03	16.91±6.89	23.27±9.53	29.57±11.61
损伤后24 h	30.18±10.82	21.52±2.89	28.41±7.07	33.54±2.31	13.45±3.37	32.02±3.16	33.49±4.46

组别	死后0 h	0 ℃		16 ℃		26 ℃
组别	死后0 h	死后12 h	死后24 h	死后12 h	死后24 h	死后12 h	死后24 h
损伤后0 h	1.00±0.25	0.76±0.20	1.50±0.31	2.20±0.50	1.52±0.15	1.87±0.35	0.72±0.15
损伤后4 h	1.97±0.46	2.76±0.68	1.58±0.18	2.21±0.36	1.79±0.42	2.31±0.41	7.02±2.08
损伤后8 h	2.53±0.50	3.51±0.83	4.33±1.16	4.00±0.99	3.64±1.31	5.10±0.97	9.84±2.50
损伤后12 h	9.20±1.23	13.87±4.51	16.75±3.32	7.98±1.83	12.52±2.45	5.52±1.69	12.49±3.47
损伤后16 h	13.71±4.14	16.57±5.46	13.38±1.80	18.33±5.13	15.56±7.90	21.37±8.30	16.64±1.88
损伤后20 h	15.48±6.27	19.75±2.31	25.71±9.46	23.23±5.03	16.91±6.89	23.27±9.53	29.57±11.61
损伤后24 h	30.18±10.82	21.52±2.89	28.41±7.07	33.54±2.31	13.45±3.37	32.02±3.16	33.49±4.46

[1]	Yong ZENG, Dong-hua ZOU, Ying FAN, Qing XU, Lu-yang TAO, Yi-jiu CHEN, Zheng-dong LI. Research Progress and Forensic Application of Human Vascular Finite Element Modeling and Biomechanics [J]. Journal of Forensic Medicine, 2023, 39(5): 471-477.
[2]	Yu-xin SUN, Xiao-juan GONG, Xiu-li HAO, Yu-xin TIAN, Yi-ming CHEN, Bao ZHANG, Chun-xia YAN. Screening of Genes Co-Associated with Sudden Infant Death Syndrome and Infectious Sudden Death in Infancy and Bioinformatics Analysis of Their Regulatory Networks [J]. Journal of Forensic Medicine, 2023, 39(5): 433-440.
[3]	Yu YANG, Fan-zhang LEI, Yu-you DONG, Jian-long MA, Qi-qiang SHI, Xue-song YE. Retrospective Analysis of Death Cases of Oral Diphenidol Hydrochloride Poisoning [J]. Journal of Forensic Medicine, 2023, 39(4): 393-398.
[4]	Qing-qing XIANG, Li-fang CHEN, Qin SU, Yu-kun DU, Pei-yan LIANG, Xiao-dong KANG, He SHI, Qu-yi XU, Jian ZHAO, Chao LIU, Xiao-hui CHEN. Research Progress on Microbial Community Succession in the Postmortem Interval Estimation [J]. Journal of Forensic Medicine, 2023, 39(4): 399-405.
[5]	Qin SU, Qian-ling CHEN, Wei-bin WU, Qing-qing XIANG, Cheng-liang YANG, Dong-fang QIAO, Zhi-gang LI. Metabonomics Analysis of Brain Stem Tissue in Rats with Primary Brain Stem Injury Caused Death [J]. Journal of Forensic Medicine, 2023, 39(4): 373-381.
[6]	Xu-dong ZHANG, Yao-ru JIANG, Xin-rui LIANG, Tian TIAN, Qian-qian JIN, Xiao-hong ZHANG, Jie CAO, Qiu-xiang DU, Jun-hong SUN. Postmortem Interval Estimation Using Protein Chip Technology Combined with Multivariate Analysis Methods [J]. Journal of Forensic Medicine, 2023, 39(2): 115-120.
[7]	Wu LONG, Peng-fei QU, Lin MA, Rui WANG, Yan-mei XI, Yu-hua LI, Sheng-jie NIE, Ting DUAN, Jin-liang DU, Xue TANG, Jing-feng ZHAO, Pu-ping LEI, Yue-bing WANG. Cytotoxicity of 4 Wild Mushrooms in a Case of Yunnan Sudden Unexplained Death [J]. Journal of Forensic Medicine, 2023, 39(2): 121-128.
[8]	Jun-wei GAO, Yang LU, Yan-jun LI, Dong-hua ZOU, Guang-long HE, Yan-bin WANG. Survey on the Construction Status of Forensic Virtual Autopsy Laboratory and the Applicability of Laboratory Accreditation [J]. Journal of Forensic Medicine, 2023, 39(2): 186-192.
[9]	Fang-Fang LIU, Hui WU, Wei WANG, Ying XIE. Changes of 1，5-AG in Vitreous Humor of Rabbit Cadavers with Hyperglycemic Metabolism [J]. Journal of Forensic Medicine, 2023, 39(1): 13-17.
[10]	Zhi-ling TIAN, Ruo-lin WANG, Jian-hua ZHANG, Ping HUANG, Zhi-qiang QIN, Zheng-dong LI, He-wen DONG, Dong-hua ZOU, Mao-wen WANG, Zhuo LI, Lei WAN, Xiao-tian YU, Ning-guo LIU. Comparison of CT Values between Thrombus and Postmortem Clot Based on Cadaveric Pulmonary Angiography [J]. Journal of Forensic Medicine, 2023, 39(1): 7-12.
[11]	Jia-yi WU, You-jia YU, Kai LI, Xin YIN, Han-ting FAN, Rong LI, Zhi-wei ZHANG, Wei TANG, Hui-jie HUANG, Feng CHEN. Report on Cardiac Gross Pathologic Measurements of Sudden Cardiac Death in Adults [J]. Journal of Forensic Medicine, 2023, 39(1): 1-6.
[12]	Hui WU, Fang-fang LIU, Jun-da WU, Ying XIE. Research Progress on Estimation of Postmortem Interval Based on Ocular Tissues Structure [J]. Journal of Forensic Medicine, 2023, 39(1): 50-56.
[13]	Tian-pu WU, Jian-long MA, Xin-biao LIAO, Dong-chuan ZHANG, Kai-jun MA, Yan-geng YU, Long CHEN. Research Progress on Molecular Changes in Pulmonary Hypoxia and Cause of Death Identification in Mechanical Asphyxia [J]. Journal of Forensic Medicine, 2023, 39(1): 57-65.
[14]	Yuan-bo KANG, Si-fan WANG, Ting-jun CHEN, Ya-dong GUO, Chang-quan ZHANG. Research Progress of Exosomes and Their Forensic Significance [J]. Journal of Forensic Medicine, 2022, 38(6): 754-762.
[15]	Yu-qing JIA, Tian-qi WANG, Rui ZHAO, Bao-li ZHU, Zhi-peng CAO. Effects of Postmortem Hemolysis and Ultrafiltration on Creatinine Detection Results [J]. Journal of Forensic Medicine, 2022, 38(6): 697-701.