Clinical effect of spleen aminopeptide on improving liver function damage and immune function in children with infant hepatitis syndrome

Abstract

BACKGROUND

Infant hepatitis syndrome (IHS) is a clinical syndrome in infants less than one year of age with generalized skin jaundice, abnormal liver function, and hepatomegaly due to various etiologies such as infection.

AIM

To investigate the effect of IHS patients, after treatment with arsphenamine-based peptides, on patients' liver function damage and immune function.

METHODS

Of 110 patients with IHS treated in our hospital from January 2019 to January 2021 were grouped according to the randomized residual grouping method, with 5 cases in each group shed due to transfer, etc. Ultimately, 50 cases remained in each group. The control group was treated with reduced glutathione, and the treatment group was treated with sesquiterpene peptide based on the control group. Observe and compare the differences in indicators after treatment.

RESULTS

The comparison of serum total bilirubin, direct bilirubin, and serum alanine transferase after treatment was significantly different and lower in the treatment group than in the control group (P < 0.05). The comparison of CD4⁺, CD3⁺, CD4⁺/CD8⁺ after treatment was significantly different and higher in the treatment group than in the control group, and the comparison was statistically significant (P < 0.05). The complication of the two groups showed that the rash, cough and sputum, elevated platelets, and gastrointestinal reactions in the treatment group were significantly lower than those in the control group, and the differences were statistically significant by test (P < 0.05).

CONCLUSION

The comparative study of IHS treated with arsphenamine combined with reduced glutathione is more effective.

Key Words: Infant hepatitis syndrome, Splenamin, Reduced glutathione, Liver function, Immune function, Complication

Core Tip: In this study, infant hepatitis syndrome (IHS) is a clinical syndrome in infants less than one year of age with generalized skin jaundice, abnormal liver function, and hepatomegaly due to various etiologies such as infection. The comparative study of IHS was better treated with splenamine combined with reduced glutathione, which effectively improved patients' liver function and immune level, reduced adverse reactions, and provided some reference value for clinical treatment of IHS.

INTRODUCTION

Infant hepatitis syndrome (IHS) is a clinical syndrome in infants less than one year of age with generalized skin jaundice, abnormal liver function, and hepatomegaly due to various etiologies such as infection[1]. Most of the jaundice appears within the first 12 months of life. It is accompanied by darkening of the urine and lightening of the stool, usually with hepatomegaly, and about 50% of patients also have enlarged spleen and liver function abnormalities, such as increased serum transaminases in laboratory tests. At the same time, children with IHS have inflammatory changes in the liver that cause metabolic disorders in the body, resulting in reduced protein synthesis and impaired immune function, which can seriously affect the average growth and development of infants and cause hepatic malnutrition, which can be complicated by multiple organ and systemic disorders, including secondary infections, rickets, and intracranial hemorrhage[2,3].

Splenaminopeptide is an immunomodulator, the main component of which is derived from peptides and nucleotides extracted from fresh pig spleen and is used in the treatment of cellular immune hypofunction, immunodeficiency and autoimmune dysfunctional diseases such as recurrent respiratory infections, bronchitis, pneumonia, asthma, severe herpes zoster, etc. It is also used in patients with malignant tumors after radiotherapy, chemotherapy, and surgery to improve the patients' autoimmunity[4]. Splenaminopeptide can inhibit the secretion of IL-4 by Th2 cells, which can enhance immunity against viruses by relieving the inhibition of lymphocytes by IL-4 and the inhibition of the ability of macrophages to phagocytose viruses[5]. By acting on three links: immune messaging, lymphocyte activation, and receptor regulation, splenaminopeptide has sufficient time to repeatedly engage with recipient lymphocytes in vivo, improving the ratio of CD4+ CD8+ cells, resulting in a significant increase in the number of lymphocytes and enhancing the cellular immune function of patients[6]. It can be seen that IHS is one of the serious diseases that threaten the health and quality of life of infants and children, and how to effectively prevent and treat this disease is a topic worthy of our in-depth study. However, whether splenamine-based peptides improve the liver function damage and immune status of patients with IHS has not been clinically reported. However, there are no clinical reports on whether splenamine improves liver function impairment and immune status in IHS patients. Based on this, we have explored the effect of splenic on liver function damage and immune function of IHS patients and analyzed the clinical efficacy. The research results are reported as follows.

MATERIALS AND METHODS

General information

The actual sample size included in this study was the 110 children who were treated in our hospital from January 2019 to January 2021, divided into control and treatment groups according to the random remainder grouping method, and 5 cases were shed in each group due to transfer, new crown epidemic, lost visits, etc. Finally, 50 cases were left in each group. Diagnostic criteria: all patients met the diagnostic criteria for IHS according to the diagnostic criteria for pediatric diseases[7]: Moderate to severe yellowing of the skin over the body, abnormal liver function, lightening of stool color and deepening of urine color; palpable right subcostal liver > 2.1 cm, serum total bilirubin (TBIL) > 171 μmol/L; direct bilirubin (DBIL) < 86.5 μmol/L. Those with serum glutathione aminotransferase (ALT) > 40 U/L, liver and spleen ultrasound: showed enlarged liver, spleen may not be enlarged. In both groups, general information such as gender and age did not affect this test (Table 1).

Table 1 Comparison of general data between the two groups (n, mean ± SD).

Group	Average age (d)	The average course of disease (d)	Patient weight (kg)	First symptoms
				Jaundice	Pneumonia	Diarrhea
Control group (50)	41.78 ± 4.32	11.34 ± 3.25	3.34 ± 0.25	36	8	6
Therapy group (50)	41.62 ± 4.66	11.31 ± 3.64	3.31 ± 0.64	35	10	5
χ2/t	0.169	0.043	-0.309	0.327
P value	0.866	0.965	0.758	0.849

Inclusion and exclusion criteria

Inclusion criteria: (1) Patients with yellow skin on the face, obscure color, increasingly aggravated, abdominal fullness, enlarged liver and spleen with hard texture, short yellow urine, constipated stool, lighter color, dark red lips or tongue with petechiae, yellow coating; (2) age within one year, gender not limited, those with disease duration beyond two weeks, all cases have not used similar Chinese medicine treatment after the onset; and (3) the selected patients were treated with splenamine-based peptide for the first time. Exclusion criteria: (1) Patients with combined infectious diseases of the respiratory tract, gastrointestinal tract, and other systems, congenital diseases, central diseases, hemorrhagic diseases, etc.; (2) patients with other causes of jaundice and liver function impairment (such as congenital biliary atresia, endocrine hepatic glycogen accumulation disease, etc.); and (3) cases with an allergy to drug components such as drugs used in this study or those with incomplete clinical data.

Methods

The control group was treatment was initiated with reduced glutathione (0.3 to 0.6 g), uniformly mixed with glucose (5%) in 50 or 100 mL and administered intravenously, one time/d, for two weeks. After two weeks, it was changed to oral dose, 30 mg/kg/d, three times/d, and continued for two weeks) for liver protection; in the treatment group, based on the control group, the oral lyophilized powder of splenamine-tide (trade name: Fucoto, produced by Zhejiang Fengan Biopharmaceutical Co.). After two weeks, it can be taken orally at home with the medicine, and it should be sealed and stored in a dark place at 2-8 ℃, and it is forbidden to take medicine when its properties change. Children in both groups were treated for four weeks. All children had Liver function tests and imaging before and after treatment.

Follow-up and observation indexes

During the treatment, we observed daily changes in heart rate, heart rhythm, respiration, pulse, weight, urine volume, jaundice, stool color, etc. We measured blood, urine, stool routine, serum TBIL, DBIL, and ALT every week and recorded the efficacy of each course of treatment in detail every day; the Beckman CytoFLEX flow cytometer detected CD4+, CD3+, CD4+/CD8+. complete regression of yellow marks, good general condition, normal liver and spleen shrinkage, normal liver function. Improvement: Jaundice was significantly reduced, the liver and spleen shrank more than before, and the liver function of some of the indicators returned to normal. Ineffective: jaundice did not subside, and there was no change in biochemical indexes. Effective rate = (cure + improvement) - 100%.

Statistical methods

The data of this study were counted using Excel, and two physicians were available for review. The selected data were normally distributed, and the data had first, and corresponding authors entered into the computer system and proofread before using the statistical software SPSS 25.0 for relevant calculations. The measurement data were expressed as mean ± SD by independent samples t-test, and the count data was expressed as a percentage (%) or integer by χ² test, with statistical P < 0.05 indicating statistically significant differences.

RESULTS

Comparison of general data

The comparison of general data such as mean age mean disease duration, mean weight of patients, and patients' first symptoms in the two groups was not statistically significantly different by t-test and chi-square test (P > 0.05) (Table 1).

Comparison of liver function levels

The liver function level of the two groups of patients before treatment was not significantly different (P > 0.05). At the same time, the serum total bilirubin, direct bilirubin, and serum alanine transferase were significantly different after treatment. The therapy group is lower than the Control group, which is statistically significant (P < 0.05) (Table 2).

Table 2 Comparison of liver function levels between the two groups (mean ± SD).

Group	TBIL (μmol/L)		DBIL (μmol/L)		Serum alanine transferase (U/L)
	Before therapy	After treatment	Before therapy	After treatment	Before therapy	After treatment
Control group (50)	172.34 ± 32.18	75.78 ± 9.32	50.34 ± 12.25	32.51 ± 9.82	79.93 ± 19.27	69.85 ± 7.04
Therapy group (50)	173.32 ± 29.63	53.62 ± 8.66	49.36 ± 13.64	16.87 ± 3.81	73.94 ± 21.23	43.37 ± 8.30
t	-0.158	12.317	0.378	10.499	1.477	17.204
P value	0.874	0.000	0.706	0.000	0.143	0.000

DBIL: Direct bilirubin; TBIL: Total bilirubin.

Comparison of immunity levels

There was no statistically significant difference between the two groups of patients in the before-therapy immune level (P > 0.05), while after treatment, CD4+, CD3+, and CD4+/CD8+ were significantly different, and the therapy group was higher than the Control group, which was statistically significant (P < 0.05) (Table 3).

Table 3 Comparison of the immune level of the two groups of patients (mean ± SD).

Group	CD4+ (%)		CD3+ (%)		CD4+/CD8+ (%)
	Before therapy	After treatment	Before therapy	After treatment	Before therapy	After treatment
Control group (50)	32.78 ± 8.32	41.34 ± 10.25	52.51 ± 10.32	62.78 ± 10.32	0.94 ± 0.25	1.67 ± 0.12
Therapy group (50)	32.62 ± 8.66	48.26 ± 10.64	51.05 ± 10.11	73.62 ± 10.66	0.96 ± 0.24	2.49 ± 0.37
t	0.094	-3.312	0.715	-5.166	-0.408	-14.907
P value	0.925	0.000	0.477	0.000	0.684	0.000

Comparison of complications

The effective rate of 98.00% of the therapy group was significantly higher than that of 76.00% of the control group, and the difference was statistically significant (P < 0.05). The complication of the two groups of patients showed that the rash, cough, sputum, elevated platelets, and gastrointestinal reactions of the therapy group were significantly lower than those of the control group, and the difference was statistically significant (P < 0.05) (Table 4).

Table 4 Comparison of clinical efficacy and complications between the two groups [n (%)].

Group	Cure	Improved	Ineffective	Effective	Skin rash	Cough and sputum	Elevated platelets		Gastrointestinal reaction
Control group (50)	24 (48.00)	14 (28.00)	12 (22.00)	38 (76.00)	2 (4.00)	2 (4.00)		1 (2.00)		3 (6.00)
Therapy group (50)	14 (28.00)	35 (70.00)	1 (2.00)	49 (98.00)	0	1 (2.00)		0		1 (2.00)
χ2				10.698						4.000
P value				0.001						0.046

DISCUSSION

The pathogenesis of IHS is complex, and in infancy, the development and progression of liver lesions are associated with interactions between various types of cells in the liver[8]. Hepatocytes, blood sinusoidal endothelial cells, bile duct epithelial cells, blastocytes, and lipid storage cells of the liver can interact and interfere with each other, either directly or through some cytokines[9]. Any damage to these cells can disrupt the normal function of other cells, leading to liver dysfunction and causing apoptosis or death of hepatocytes, resulting in different pathologies and regressions. IHS varies with various etiologies[10]. In viral infections, most of the hepatocytes are either directly or immunologically damaged by the virus; many hepatocytes undergo lesions, necrosis, and apoptosis[11]. Bacterial infections are primarily seen in sepsis and urinary tract infections, which damage hepatocytes. IHS is caused by various metabolic disorders, in which hepatocytes are usually damaged by abnormal toxic metabolic intermediates[12]. IHS, caused by intrahepatic bile duct development disorders, first causes biliary stasis in the liver, affecting the nutrient metabolism of hepatocytes and producing lesions[13]. The treatment of common IHS can be based on the condition of intravenous glycopyrrolate, glutathione, vitamin injection, and other drugs to protect the liver and bile to reduce yellowing. Those with scleral skin yellowing are given oral Injasmine oral solution, and phenobarbital can be applied orally, which improves and increases the enzyme activity and promotes bile excretion[14]. At the same time, antiviral therapy was given to those who tested positive for pathogenesis, and antiviral agents such as adenosine, ganciclovir intravenous, and interferon intramuscular injection were used for antiviral treatment[15].

In this study, the comparison of serum total bilirubin, direct bilirubin, and serum alanine transferase after treatment of IHS with arsphenamine-combined with reduced glutathione was significantly different and lower in the treatment group than in the control group, indicating that IHS was better treated with arsphenamine-combined with reduced glutathione, which effectively improved the liver function of the patients. Glutathione is a triglyceride containing Y- amide bond and sulfhydryl group, composed of glutamic acid, cysteine, and glycine, and the sulfhydryl group on cysteine is the active group of glutathione, which binds to free radicals and scavenges them from the body through oxidative dehydrogenation[16]. Combined with heavy metals, iodoacetic acid, and mustard gas, it converts harmful toxins of the organism into harmless substances. It excretes them from the body, which has a detoxifying effect, and glutathione improves the organism's immunity, making glutathione have a significant hepatoprotective effect[17]. The oral lyophilized powder of splenamine-based peptides can effectively improve the immune function of the body and enhance the antiviral ability of the body, which plays a synergistic role in treating IHS.

In this study, the differences in CD4+, CD3+, and CD4+/CD8+ after treatment were significant and higher in the treatment group than in the control group, indicating that IHS is better treated with arsphenamine-combined with reduced glutathione, which effectively improves the immune level of patients. T lymphocytes are divided into subpopulations according to their phenotypes and functions, namely, killer T cells, suppressor T cells, late metaplastic reactive T cells and inducible helper T cells[18]. The former two are CD8+ subpopulations, and the latter two are CD4+ subpopulations, which induce and regulate each other, forming a T cell regulatory network that exerts its killing effect on target cells and its positive and negative feedback regulation of immune response processes, such as anti-infection and anti-tumor immunity[19]. IHS causes dysfunctional T-cell subpopulations in the organism. The CD4+/CD8+ cell ratio decreases, and the immune regulatory network consisting of various immune cells and cytokines is imbalanced, forming a pathological vicious circle that leads to the development and progression of liver disease[20]. It has been shown that HBV can invade the peripheral blood lymphocytes of patients and affect the normal functioning of cellular functions. The lack of IL-2 in the body directly affects the normal functioning of killer T cells[21]. The low antiviral ability of the body makes HBV infect the body for a long time, on the other hand, it aggravates the autoimmune reaction in the body and aggravates the disease, while splenamine-based oral lyophilized powder has the effect of regulating immunity and inhibiting viral replication, and has better efficacy in treating chronic hepatitis B. It is easy to take and has few adverse effects, which provides a new way and method for the treatment of chronic hepatitis[22].

Although this study has some novelty, there are also shortcomings. The clinical efficacy of tegretolide-reduced glutathione treatment on IHS is significant, but the specific mechanism has not been studied in depth for a long time. The cases collected from the same hospital could have been more representative, and the exclusion and inclusion were subjective, which may lead to biased results.

CONCLUSION

In conclusion, the comparative study of IHS was better treated with splenamine combined with reduced glutathione, which effectively improved patients' liver function and immune level, reduced adverse reactions, and provided some reference value for clinical treatment of IHS.

ACKNOWLEDGEMENTS

The authors thanks to the help from institutions and families, also the cooperation of all participants and their families.