Effects of Long-term High-dose Induction of Doxycycline on Liver and Kidney Functions of Diannan Miniature Pigs

doi:10.13560/j.cnki.biotech.bull.1985.2024-0882

Abstract

Abstract:

Objective To explore the impact of continuous high-dose intake of doxycycline (Dox) in the Tet-On system on the health status of experimental pigs. Method Nine 6-month-old Diannan miniature pigs were selected as model animals, and the recommended dose groups of 3 (30 mg/kg) times and 5 (50 mg/kg) times and blank control group were set up. The experimental pigs in each group were intramuscularly injected with corresponding drugs or physiological saline for 30 d. During the experiment, blood was collected on the day 0, 10, 20, and 28 for blood routine and liver and kidney function index measurements. The experimental pigs’ livers and kidneys at 2 h after last injection were subjected to morphological observation, pathological examination and disease detection. Result Compared with the control group, Lym# and MPV decreased, while PLT increased in the 30 mg/kg dose group; while the changes in these indicators were more significant in the 50 mg/kg dose group, and RDW-SD significantly reduced, meanwhile ALT, MG, P and other indicators increased, and NA, CO2, CYSC and other indicators decreased, showing more obvious damages to liver and kidney function. Histopathological examination results showed that the livers and kidneys of the experimental pigs in the 30 mg/kg dose group and the 50 mg/kg dose group had varying degrees of lesions, and the liver inflammatory cells and red blood cell infiltration in the 50 mg/kg dose group were more severe. The kidneys showed more obvious atrophy and structural tissue damage. The pathogenic infection detection results excludes the interference of viral infection on the experimental results. Conclusion This study preliminarily proves that long-term use of high-dose Dox can cause significant damage to experimental pigs' liver and kidney functions, and the degree of damage is positively correlated with the dose of Dox. This study may provide references for the research of conditional induced expression in large gene editing animals.

Key words: doxycycline hydrochloride, Tet-On system, Diannan miniature pigs, liver and kidney injury

LOU Ya-nan, XIONG Jing-jing, LOU Ya-ling, WANG Fu-bin, CHENG Wen-jie, CHEN Xiao-jing, LI Jia-yu, PU Shao-xia, WEI Hong-jiang, ZHAO Hong-ye. Effects of Long-term High-dose Induction of Doxycycline on Liver and Kidney Functions of Diannan Miniature Pigs[J]. Biotechnology Bulletin, 2025, 41(4): 345-354.

Figures/Tables 11

Table 1 Grouping situation of experimental pigs

组别 Group	序号 Serial number	耳号 label	体重 Weight/kg	剂量 Dose/mL
对照 Control	1	2A016S3	17.00	17.00
	2	3072T1	14.80	27.80
	3	2793T1	29.00	29.00
30 mg/kg	1	3152PA3	12.00	7.20
	2	3045T1	22.80	13.68
	3	3057Q1	13.00	7.80
50 mg/kg	1	3149PA3	13.00	13.00
	2	2A051T1	27.80	27.80
	3	2A073P1	18.00	18.00

Table 2 Primer sequences for pathogenic infection detection

引物名称	引物序列
Primer name	Primer sequence（5'-3'）
PEDV-F	GCGTAGTTGAGATTGTTG
PEDV-R	CCTCTGTTGTTACTTGGA
TGEV-F	GATGGAGTTGTCTGGGTTGC
TGEV-R	GTGACCTTGAATTGTCCCTT
RV-F	TATTTCCGCAAGCRCCRCCATT
RV-R	ACCHGGTGGAAATACYGGT-CCT
MP-1-F	AGCCTAGCTACCGATTGGCT
MP-1-R	CTGGGTAACTGACTTTGCAA

Fig. 1 Experimental pig survival curve

Fig. 2 Effects of Dox on the blood physiological indicators of experimental pigsA: Experimental pig blood Lym# count. B: Experimental pig blood MPV. C: Experimental pig blood PLT count.D: Experimental pig blood RDW-SD. E: Experimental pig blood EOS # count. *P<0.05, **P<0.01, ##P<0.01

Fig. 3 Effects of Dox on related indicators of experimental pig liver functionA: Prealbumin (PA). B: Cholinesterase (PCHE). C: Ratio of albumin and globulin (A/G). D: Serum alanine aminotransferase (ALT). E: Aspartate aminotransferase (AST). F: Ratio of serum glutamic propyl to glutamic and grass content (L/S). *P<0.05, **P<0.01

Fig. 4 Effect of Dox on the related indicators of renal function in experimental pigsA: Cystatin C (CYSC); B: Creatinine (CREA). *P<0.05, #P<0.05

Fig. 5 Effects of Dox on the blood electrolyte related indicators of experimental pigsA: Magnesium ion (MG) . B: Inorganic phosphorus (P). C: Bicarbonate (CO2). D: Sodium ion (NA). *P<0.05, #P<0.05

Fig. 6 Effects of Dox on experimental pig liver and kidney organsA: Behind the left ear of experimental pigs in 30 mg/kg group. B: Experimental pig liver in 30 mg/kg group. C: 30 mg/kg experimental pig kidney. D: Cross-section of pig kidney in 30 mg/kg group. E: Behind the left ear of experimental pigs in 50 mg/kg group. F: 50 mg/kg experimental pig liver. G: Pig kidney in 50 mg/kg group. H: Kidney cross-section of experimental pigs in the 50 mg/kg group

Fig. 7 Histopathological examination of experimental pig's liver and kidneyA: Experimental pig liver in control group. B: Experimental pig liver in 30 mg/kg group. C: 50 mg/kg experimental pig liver. D: Control group experimental pig kidney. E: Pig kidney in 30 mg/kg group. F: Experimental pig kidney in 50 mg/kg group

Fig. 8 Detection of related diseases in experimental pigs in the 30 mg/kg group on the 28 th day after administrationA: Epidemic diarrhea (459 bp). B: Transmissible gastroenteritis (517 bp). C: Rotavirus (289 bp). D: Mycoplasma pneumonia (457 bp). The same below

Fig. 9 Detection of related diseases at the death of experimental pigs in the 30 mg/kg group

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