Oral Administration of Bovine Blood Peptide Generated No Adverse

Effect on Healthy Rats

Zhenghao Bao

, Yuhao Zhang

, Ao Wang

, He Huang

, Yuexin Xu

, Hongpeng He

1, * f

Aqin Wang

3,* g

and Jun Yu

3,* h

Key Laboratory of Industrial Microbiology, Ministry of Education and Tianjin City, State Key Laboratory of Food

Nutrition and Safety, College of Biotechnology, Tianjin University of Science and Technology, 300457, Tianjin, China

Department of Pathology, Mentougou Hospital in Beijing, 102300, Beijing, China

Hangzhou BIBAU Biotechnology Co. Ltd, 310016, Hangzhou, China

Keywords: IDA, Rat, Bovine Blood Peptide, Heme Iron, Iron Supplement.

Abstract: IDA (iron deficiency anemia) is a disease with high incidence in many countries. Inorganic iron supplements,

typically ferrous sulfate, are widely utilized in the prevention and treatment of IDA. However, the

bioavailability of inorganic iron is only 2% to 20%. In addition, the gastrointestinal side effects occur

frequently. Heme iron which is isolated from animal blood is a promising choice for IDA patients. Before

extensive utilization of heme iron in clinic, safety evaluation is indispensable. In this study, with untreated

rats and ferrous sulfate-treated rats as controls, bovine blood peptide which carries heme iron was applied to

normal female rats. After 30 days of gavage feeding, no significant difference in body weight and organ

coefficient was observed. Serological examination revealed that oral administration of bovine blood peptide

did not disrupt iron metabolism nor caused adverse effects on liver and kidney functions. Pathological HE

staining of gastrointestinal tract showed that bovine blood peptide induced much less inflammatory irritation

than ferrous sulfate. These results suggest that bovine blood peptide is a kind of safe and reliable organic iron

supplement for IDA patients.

1 INTRODUCTION

Iron is one of the trace elements needed by the human

body. Due to some congenital or acquired factors, the

amount of stored iron in the body is too low to support

the synthesis of functional iron (hemoglobin, etc.)

and consequently resulted in iron deficiency anemia

(IDA) (Lin 2013). The pathogenic factors of IDA

include excessive iron loss, iron utilization disorder,

iron uptake deficiency, iron malabsorption, iron

transport disorder (Chen 2013). As reported, about

one billion people worldwide have some form of iron

deficiency, and more than half of them have

developed IDA (WHO 2001). The WHO report in

2011 mentioned that the prevalence of IDA in

children in China was 7.8% (WHO 2011). The

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prevalence of IDA is more than 20% in women

(Zhang 2010). Generally, Infants, growing children

and women of childbearing age are vulnerable to IDA

(Liu 2012, Dalhøj 1991). Iron deficiency not only

leads to decreased red blood cells but also weaken the

activity of iron-containing enzyme in cells and

triggers clinical symptoms of IDA (Ge 2013).

Supplementation of iron increases the level of

hemoglobin (Hb). At present, there are two types of

iron supplements for IDA treatment (Ulas 2010). Oral

ferrous sulfate is the main drug used in clinical

treatment of IDA because of its low price and

significant effect. But side effects, such as irritation

of the gastrointestinal tract, constipation, or diarrhea

cannot be ignored. In addition, it has low

bioavailability (He 1995). Studies have shown that

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Bao, Z., Zhang, Y., Wang, A., Huang, H., Xu, Y., He, H., Wang, A. and Yu, J.

Oral Administration of Bovine Blood Peptide Generated No Adverse Effect on Healthy Rats.

DOI: 10.5220/0011368100003444

In Proceedings of the 2nd Conference on Artiﬁcial Intelligence and Healthcare (CAIH 2021), pages 313-317

ISBN: 978-989-758-594-4

313

excessive iron intake can cause changes in intestinal

microbiota and oxidative stress injury in the body

(Alexeev 2017, Zhang 2017).

The most abundant protein in blood is

hemoglobin which can be purified and processed to

produce heme iron supplements. Heme iron is better

than inorganic iron. Bovine blood peptide also

provides human body with useful peptides and amino

acids. Therefore, it is a potential substitute of ferrous

sulfate. The advantages of blood peptide and heme

iron have been extensively reported while the safety

and reliability remain to be further clarified.

This study aimed to evaluate the safety of bovine

blood peptide in normal female rats. Results of the

present study will provide useful information for the

clinical application of bovine blood peptide in IDA

treatment.

2 MATERIALS AND METHODS

2.1 Animals and Reagents

Forty-eight female SD rats with body weight between

60-80g (purchased from SiPeiFu, Beijing, China)

were randomly divided into 6 groups, namely control

group, bovine blood peptide low-dose group (2

mg/kg/d), bovine blood peptide medium dose group

(20 mg/kg/d), bovine blood peptide high-dose group

(100 mg/kg/d), ferrous sulfate low-dose group(100

mg/kg/d) and ferrous sulfate high-dose group(200

mg/kg/d). Bovine blood peptide powder and ferrous

sulfate tablets were provided by BIBAU, Hangzhou,

China. After adaptive feeding for 5 days, rats were

continuously feed with different iron supplements as

indicated by gavage for 30 days.

2.2 Measurement of Blood

After gavage administration, rats were sacrificed and

blood was collected for the measurement of liver and

kidney functions with biochemical or ELISA kits

(from Jiancheng, Nanjing, China) on an automatic

biochemical analyzer.

2.3 Pathological Examination

Stomach tissues were collected, embedded with

paraffin and then sliced for HE staining to examine

tissue damage and the infiltration of inflammatory

cells.

2.4 Statistical Analysis

All data were expressed as mean ± standard

deviation. Prism 8.0 was used for statistical analysis.

One-way analysis of variance (ANOVA) with P <

0.05 was used to consider the difference statistically

significant.

3 RESULTS

3.1 Body Weight of Rats Was Not

Affected

The body weight of rats after 30 days of iron

supplementation was shown in Fig. 1. At the end of

the experiment, the body weight of rats in each group

was about 200g, and there was no significant

difference among different groups (P > 0.05). The

results showed that bovine blood peptide did not

affect the normal growth of rats, which was consistent

with the results of traditional iron supplementation

agent ferrous sulfate.

Figure 1: Rat body weight post 30-day feeding.

3.2 Liver and Kidney Organ

Coefficients and Functions Were

Unaffected

To evaluate the safety of bovine blood peptide, size

of liver and kidney was measured. As shown in Table

1, compared with the blank control, liver and kidney

organ coefficients of various experimental groups

were not significantly different (P > 0.05). By

detecting ALT, bilirubin and Glo (Figure 2), it was

found that there was no significant difference in liver

function in each group supplemented with iron (P >

0.05). The results showed that the functions of liver

and kidney were not affected by bovine blood peptide

or ferrous sulfate.

CAIH 2021 - Conference on Artiﬁcial Intelligence and Healthcare

314

Table 1: Organ coefficient of rats fed with iron supplements.

Group Or

an coefficient

(

/100

)

, mean ± SD

liver kidney

Blank control group 4.67±0.47 0.57±0.06

Blood peptide low dose group 4.92±0.25 0.69±0.03

Blood peptide medium dose group 4.57±0.33 0.71±0.08

Blood peptide high dose group 4.76±0.41 0.58±0.05

Ferrous sulfate low dose group 4.87±0.27 0.51±0.08

Ferrous sulfate high dose group 4.75±0.14 0.46±0.01

A. Alanine aminotransferase, B. total bilirubin, C. globulins

Figure 2: Rat liver function.

3.3 Glycolipid Metabolism Were Not

Affected by Iron Supplements

Bovine blood peptide used in this study was a mixture

of peptide in different length prepared from blood

protein digestion. It was previously reported that

some peptide might affect blood glucose. To test

whether the blood glucose and lipid of normal rats

altered after ingestion of bovine blood peptide, rat

sera were examined. There was no significant

difference in serum glucose, total cholesterol and

triglyceride levels among groups supplemented with

iron and the control group (Figure. 3) (P > 0.05). The

results showed that bovine blood peptide did not

affect the glycolipid metabolism of normal rats after

intragastric administration.

Oral Administration of Bovine Blood Peptide Generated No Adverse Effect on Healthy Rats

315

A. glucose, B. total cholesterol, C. triglyceride

Figure 3: Rat Glucose and Lipid Metabolism Index

3.4 Iron Metabolism Was Not Affected

by Iron Supplements

To test whether bovine blood peptide would increase

the iron concentration in serum in normal rats, serum

iron was determined. As show in Figure 4, there was

no difference in serum iron concentration among

different groups (P > 0.05), and there was no

significant dose-response relationship. The results

indicate that the supplementation of blood peptide

iron or ferrous sulfate did not significantly change the

status of iron metabolism in healthy rats fed normal

diet.

Figure 4: The serum iron content of rats in different groups.

3.5 Bovine Blood Peptide Caused Less

Gastrointestinal Side Effects than

Ferrous Sulfate

A major disadvantage of ferrous sulfate is severe

gastrointestinal side effects after taken orally. To

determine the effects of iron supplements on

gastrointestinal tract, HE staining was performed.

Compared with the normal control group, the

number and distribution of inflammatory cells in the

low dose and medium dose blood peptide groups were

not obvious (Figure 5). In the high-dose group, the

mild inflammatory reaction was mainly in the mucosa

layer, and there was no increase of inflammatory cells

in the submucosa. Whereas gastric tissues from low

and high dose ferrous sulfate groups displayed

infiltration of neutrophils and lymphocytes in the

mucous and the submucosa layers. In ferrous sulfate

high dose group, interstitial edema and vascular

dilatation and congestion were observed. These results

demonstrated that bovine blood peptide is less

irritating to the gastrointestinal tract than ferrous

sulfate.

Figure 5: Changes in the stomach tissue in rats (HE

staining, 100×): (A) blank control; (B) blood peptide low

dose; (C) blood peptide medium dose; (D) blood peptide

high dose; (E) ferrous sulfate low dose; (F) ferrous sulfate

high.

4 CONCLUSIONS

In this study, the body weight, liver and kidney

function, liver and kidney size, glucose and lipid

metabolism, and iron metabolism of normal rats were

not significantly affected by the supplementation of

bovine blood peptide or ferrous sulfate. Different

from ferrous sulfate, bovine blood peptide didn’t lead

to obvious inflammatory change in gastrointestinal

mucosa. Histological examination demonstrated that

compared with ferrous sulfate, bovine blood peptide

is a type of safer iron supplement for the treatment of

IDA and can avoid the gastrointestinal irritation

caused by inorganic iron supplements.

CONFLICT OF INTEREST

We have no conflicts of interest to disclose.

CAIH 2021 - Conference on Artiﬁcial Intelligence and Healthcare

316

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