Preliminary Study on Construction Method of Type 1 Diabetes Model in Banna Miniature Pig

doi:10.12300/j.issn.1674-5817.2024.060

Abstract

Abstract:

Objective To construct a Type 1 diabetes model in miniature pigs and explore postoperative care strategies for effectively prolonging the survival time of the model pigs. Methods Seven Banna miniature pigs were selected for pancreatectomy. Glucose, vitamins, and antibiotics were administered for 3-5 days after surgery to aid recovery. Blood glucose and urine glucose levels were measured twice a day in the morning and evening to adjust insulin supplementation accordingly. The model pigs were observed daily and records were kept, including orexis, psychosis, weakness, skin ulcer, and feces and urine. Body weight was measured weekly until the death of the model animals. Based on the model pigs' condition, glucose injection and Ringer's lactate solution were administered to supplement nutrition and correct electrolyte imbalances. Results All seven Banna miniature pigs showed typical symptoms of diabetes: random blood glucose levels higher than 11.1 mmol/L after pancreatectomy, far exceeding the average blood glucose level of 6.0 mmol/L in normal pigs; positive urine glucose; and progressive weight loss. These features indicated the successful construction of Type 1 diabetes model. Additionally, Type 1 diabetic pigs that survived more than 8 weeks showed progressive hair loss and skin ulceration. Euthanasia was performed on model pigs when they were unable to stand or even eat independently, and pathological examination and HE staining were conducted on tissues collected from affected organs such as the liver, kidneys, and skin. Pathological sections revealed liver congestion, massive glycogen accumulation, ballooning degeneration of hepatocytes, and progressive liver fibrosis, along with glomerular congestion, vacuolar degeneration in renal tubular epithelial cells, proteinuria, dermal congestion, thinning of vascular walls, and varying degrees of parakeratosis and dyskeratosis in the liver, kidneys, and skin tissues due to prolonged hyperglycemia. The average survival time of the constructed Banna miniature pig diabetes model was 44 d, with a maximum survival time of 121 d. Conclusion Type 1 diabetes model can be constructed successfully in Banna miniature pigs through pancreatectomy. With meticulous postoperative care, a long-term Type 1 diabetes model with significant complications can be achieved, providing a stable large-animal model for Type 1 diabetes treatment strategies.

Key words: Banna miniature pig, Type 1 diabetes model, Pancreatectomy, Postoperative care, Complications of diabetes

CLC Number:

R587.1

LIU Kai,HU Yuqi,GENG Yatian,et al. Preliminary Study on Construction Method of Type 1 Diabetes Model in Banna Miniature Pig[J]. Laboratory Animal and Comparative Medicine, 2024, 44(4): 384-392. DOI: 10.12300/j.issn.1674-5817.2024.060.

Figures/Tables 6

Table 1 Scoring table for post-pancreatectomy status of Banna miniature pigs

观察指标 Observation indicators	评分 Score
观察指标 Observation indicators	0	1	2	3	4	5
饮食欲 Orexis	饲料无采食痕迹，拒食	饲料有采食痕迹，但未见明显减少	饲料有采食痕迹且减少约1/3	饲料有采食痕迹且减少约2/3	料槽无剩余饲料，加料时有采食意愿但不强烈	料槽无剩余饲料，且加料后采食意愿强烈
精神状态 Psychosis	精神沉郁，目光呆滞无神，对外界刺激无明显反应，不愿与人互动	精神较沉郁，对外界刺激有轻微反应，互动时反应迟钝	精神较沉郁，对外界刺激有一定的反应，互动时反应稍迟钝	精神较正常，对外界刺激有正常的反应，互动时反应速度正常	精神较亢奋，对外界刺激反应较激烈，互动时反应较灵敏	精神亢奋，对外界刺激敏感，互动时反应积极迅速
虚弱程度 Weakness	活动意愿强烈，趴卧较少，无需驱赶或引诱即可观察到站立活动	除采食等生理活动外，有一定的活动意愿，驱赶或引诱时可迅速站立，无需辅助站立	除采食等生理行为外，还具有一定的活动意愿，驱赶或引诱时可站立，无需辅助站立	较喜卧，除采食等生理需求外不愿站立活动，驱赶或引诱时可自主站立移动，但仍表现为不愿站立，辅助站立后可支撑并正常行走，后自行趴卧	喜卧，驱赶或引诱时不愿站立，辅助站立后可勉强支撑，后摔倒	喜卧，驱赶或引诱均不站立，辅助站立后立即摔倒，无法自主维持站立
皮肤破损情况 Skin ulcer	皮肤未见破损	皮肤破损面积约10%	皮肤破损面积约20%	皮肤破损面积约30%	皮肤破损面积约40%	皮肤破损面积约50%及以上
粪尿量 Feces and urine	笼具中未见粪便、尿液残留，且未观察到膀胱有尿液蓄积或包皮憩室充盈	笼具中见极少量粪便、尿液，且未观察到膀胱有尿液蓄积或包皮憩室充盈	笼具中见少量粪便、尿液，且未观察到膀胱有尿液蓄积或包皮憩室充盈	笼具中见一定量粪便、尿液，偶尔能观察到排便、排尿	笼具中见较多量粪便、尿液，经常能观察到排便、排尿	笼具中见大量粪便、尿液，总是能观察到排便、排尿

Table 1 Scoring table for post-pancreatectomy status of Banna miniature pigs

观察指标 Observation indicators	评分 Score
观察指标 Observation indicators	0	1	2	3	4	5
饮食欲 Orexis	饲料无采食痕迹，拒食	饲料有采食痕迹，但未见明显减少	饲料有采食痕迹且减少约1/3	饲料有采食痕迹且减少约2/3	料槽无剩余饲料，加料时有采食意愿但不强烈	料槽无剩余饲料，且加料后采食意愿强烈
精神状态 Psychosis	精神沉郁，目光呆滞无神，对外界刺激无明显反应，不愿与人互动	精神较沉郁，对外界刺激有轻微反应，互动时反应迟钝	精神较沉郁，对外界刺激有一定的反应，互动时反应稍迟钝	精神较正常，对外界刺激有正常的反应，互动时反应速度正常	精神较亢奋，对外界刺激反应较激烈，互动时反应较灵敏	精神亢奋，对外界刺激敏感，互动时反应积极迅速
虚弱程度 Weakness	活动意愿强烈，趴卧较少，无需驱赶或引诱即可观察到站立活动	除采食等生理活动外，有一定的活动意愿，驱赶或引诱时可迅速站立，无需辅助站立	除采食等生理行为外，还具有一定的活动意愿，驱赶或引诱时可站立，无需辅助站立	较喜卧，除采食等生理需求外不愿站立活动，驱赶或引诱时可自主站立移动，但仍表现为不愿站立，辅助站立后可支撑并正常行走，后自行趴卧	喜卧，驱赶或引诱时不愿站立，辅助站立后可勉强支撑，后摔倒	喜卧，驱赶或引诱均不站立，辅助站立后立即摔倒，无法自主维持站立
皮肤破损情况 Skin ulcer	皮肤未见破损	皮肤破损面积约10%	皮肤破损面积约20%	皮肤破损面积约30%	皮肤破损面积约40%	皮肤破损面积约50%及以上
粪尿量 Feces and urine	笼具中未见粪便、尿液残留，且未观察到膀胱有尿液蓄积或包皮憩室充盈	笼具中见极少量粪便、尿液，且未观察到膀胱有尿液蓄积或包皮憩室充盈	笼具中见少量粪便、尿液，且未观察到膀胱有尿液蓄积或包皮憩室充盈	笼具中见一定量粪便、尿液，偶尔能观察到排便、排尿	笼具中见较多量粪便、尿液，经常能观察到排便、排尿	笼具中见大量粪便、尿液，总是能观察到排便、排尿

Figure 1 Results of blood glucose (A) and urine glucose (B) in Banna miniature pigs after pancreatectomyNote：Pig 1 and Pig 2 refer to the post-pancreatectomy model pigs numbered 1 and 2, respectively. Each point represents one measurement. At 7 weeks and 11 weeks, abnormal conditions were observed in the experimental animals, and supportive treatment was provided. It was difficult to collect blood samples that met the requirements of the measurement during this time, but monitoring continued once they improved. The results showed that the blood glucose concentration in the model pigs after pancreatectomy was far beyond the normal range for most of the time (the orange fine dashed line represents the mean blood glucose concentration of pigs in control group at 6.0 mmol/L, while the red thick sparse line represents the threshold for random blood glucose concentration at 11.1 mmol/L). This suggested that the diabetic model was successfully constructed.

Figure 2 Survival time of Banna miniature pigs after pancreatectomyNote：A, Survival time of 7 Banna miniature pigs after pancreatectomy, with the numbers in the figure indicating the specific survival days; B, Survival curves of two groups of Banna miniature pigs during post-pancreatectomy operative care, comparing those that received vitamin supplementation with those that did not.

Figure 3 Appearance changes of Banna miniature pigs at different times after pancreatectomyNote：A, One week post-operation for pig 1; B, Four weeks post-operation for pig 1; C, Fifteen weeks post-operation for pig 1.

Figure 4 Changes in body weight and status scores of Banna miniature pigs after pancreatectomyNote： A, Changes in body weight; B, Scores of orexis; C, Scores of skin ulcer; D, Scores of feces and urine; E, Scores of weakness; F, Scores of psychosis. Figure A shows model pig 1 and 2, while figure B-F show model pig 1. The red line represents the trend line of each indicator.

Figure 5 Gross examination and HE staining results (×400) of liver, kidney, and skin tissues in Banna miniature pigs after pancreatectomyNote：Pig 1 and 2 were diabetic Banna miniature pigs with the longest survival time after pancreatectomy. Pigs in the control group were not subjected to operation or any treatment. R, Red blood cells; F, Fibrosis; G, Glycogen; Pu, Proteinuria; Pk, Parakeratosis.

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