The aim of this study was to determine the blood routine and biochemical indices of Alagille syndrome (ALGS) and to explore the changes of the indicators of neonatal intrahepatic cholestasis caused by ALGS. This study was conducted on data of thirty-two neonatal intrahepatic cholestasis caused by ALGS treated from June 2017 to January 2020 and were randomly selected as a study group (ALGS group). Thirty-six children with unexplained cholestasis in the same period were selected as a control group. The incidences of cardiovascular malformations, special faces, vertebral changes and ocular abnormalities in the ALGS group were greater than those in the control group before and after treatment (P<0.05). There was no significant difference in WBC, RBC and PLT count and absolute value of neutrophil (Neu) between two groups before and after treatment (P>0.05). The absolute value and percentage of Neu and the ratio of Neu to lymphn ocytes (LYM) in the ALGS group were raised compared to the control group before and after treatment (P<0.05). There was no significant difference in the levels of GOT, TG, HDL, and Alb between the two groups at before and after treatment (P>0.05). The levels of γ-GT, DBil, TBA, GPT, TC, LDL, TP and GLB in the ALGS group were raised compared to the control group at before and after treatment (P<0.05). The various clinical manifestations, haematological and biochemical changes are of great significance over time in judging neonatal intrahepatic cholestasis caused by ALGS.
Short Communication · Volume 56, 25 March 2025 · Open Access
Haematological and Biochemical Indices of Neonatal Intrahepatic Cholestasis Caused by Alagille Syndrome
Affiliations & Notes ⌄
- 1Medical Office, West Hospital, Wuhan Children’s Hospital, Wuhan 430000, Hubei Province, China. * Corresponding author: yuanxilan326@163.com
Article Info ⌄
- Journal:
- Pakistan Journal of Zoology
- Published:
- 25 March 2025
From the article
- From the 162 neonatal intrahepatic cholestasis caused by ALGS treated, 68 patients were included in the study.
- For clinical manifestations, cholestasis was found in both groups, and the incidences of acute cardiovascular disease, special face, vertebral changes and ocular abnormalities in the ALGS group were raised than those in the control group after the treatment (P0.05).
- The Neu and the ratio of Neu to Lym were raised in the ALGS group than those in the control group (P0.05).
Selected from the article's own text.
Abstract
Materials and methods
Thirty-two cases of neonatal intrahepatic cholestasis caused by ALGS treated in Wuhan Children’s Hospital from June 2017 to January 2020 were randomly selected as the study group (ALGS group). All patients who signed the informed consent form, met the diagnosis and treatment criteria of neonatal intrahepatic cholestasis newborns or infants, andwith clinical manifestations of intrahepatic cholestasis were included in the study. Those with a biliary atresia detected by B-ultrasound or nuclear magnetic resonance, with toxic liver disease caused by breast milk jaundice, congenital choledochal cyst, liver tumor and other organs of toxic drugs were excluded. Thirty-six children with cholestasis of unknown origin in the same period were selected as the control group. Blood samples were collected before and after treatment. The serum was separated by centrifugation.
Blood parameters white blood cell (WBC), red blood cell (RBC), platelet (PLT), lymphocyte (Lym), Neu/Lym ratio and monocyte (MO) were analyzed by automatic blood routine analyzer (Wuhan Kelda Medical Technology Co., Ltd., model: RT-7200).
The blood serum was analyzed by automatic biochemical analyzer (Beijing Tailin Oriental Trading Co., Ltd., model: 7600) for measuring γ-glutamyl transpeptidase (γ-GT), conjugated bilirubin (DBil), total bile acid (TBA), glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT), total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL), high density lipoprotein (HDL), total protein (TP), albumin (Alb), globulin (GLB).
SPSS22.0 software package was used to analyze the statistical data. The counting data were all tested by χ²test. Analysis of variance was used to compare the measurement data between groups, and t-test was used to compare the measurement data within groups. P<0.05 was considered statistically significant.
Results and discussion
From the 162 neonatal intrahepatic cholestasis caused by ALGS treated, 68 patients were included in the study. The gender distribution was 26 female and 42 male patients aged between 0.85 and 1.60 years, with a M(SD) age of 1.23(0.41) years. Supplementary Table I shows that there are no significant differences between the two groups.
For clinical manifestations, cholestasis was found in both groups, and the incidences of acute cardiovascular disease, special face, vertebral changes and ocular abnormalities in the ALGS group were raised than those in the control group after the treatment (P<0.05) (Table I). There were statistically significant differences in cardiovascular malformation, special face, vertebral body change and ocular abnormality among groups after the treatment.
Table II shows the hematological parameters and biochemical indexes of the two groups before and after treatment. There was no significant difference over time in WBC, RBC, PLT count and MO count between the two groups (P>0.05). The Neu and the ratio of Neu to Lym were raised in the ALGS group than those in the control group (P<0.05). Moreover, there was no significant difference over time in the levels of GOT, TG, HDL and Alb between two groups (P>0.05). The levels of γ-GT, DBil, TBA, GPT, TC, LDL, TP and GLB in the ALGS group were raised over time than those in the control group (P<0.05).
Table I. Clinical manifestations related indices between groups after treatment.
| Index | Control group (n=36) | ALGS group (n=32) | χ² | P |
| Manifestation of cholestasis | 36 | 32 | - | 1.000 |
| Cardiovascular malformation | 6(16.67) | 24(75.00) | 12.285 | <0.001 |
| Special face | 15(41.67) | 25(78.13) | 5.347 | 0.021 |
| Vertebral body change | 3(8.33) | 17(53.13) | 7.243 | 0.007 |
| Ocular abnormality | 3(8.33) | 17(53.13) | 7.243 | 0.007 |
| Kidney injury | 6(16.67) | 4(12.50) | 0.129 | 0.720 |
| Acidosis | 21(58.33) | 14(43.75) | 0.744 | 0.388 |
| Underdevelopment | 30(83.33) | 24(75.00) | 0.345 | 0.557 |
For details of group, see Table I.
Hepatocytes are a kind of polar cells, which can be divided into hepatic sinusoid surface and bile tubule surface. There is an important molecular pathway regulating bile secretion and excretion on the hepatic sinusoid surface of the hepatocyte membrane, which can pump bilirubin and other bile components into the hepatocytes . In the study of animal cholestasis, some scholars have found that the expression of bile transporter in specific hepatocytes is decreased or missing in animal cholestasis model, and these abnormalities damage the function of bile transporter protein and lead to the decrease of bile flow. and then develop into cholestasis . Cholestatic liver disease has a wide range of etiology, and the diagnosis requires detailed medical history, comprehensive clinical examination and laboratory testing. The etiology of cholestasis is gradually clear, and the proportion of genetic metabolic diseases such as ALGS is gradually increasing. It is often characterized by neonatal cholestasis, and many systems such as heart, bone, eye, face and kidney can be involved, which threatens the health of patients .
Patients with neonatal intrahepatic cholestasis caused by ALGS often have typical deformities such as butterfly vertebrae, saddle nose, pointed chin, sunken eyes etc. In addition, there are many kinds of eye dysfunction, among which the posterior corneal embryonic ring is the
Table II. Haematological and biochemical indices of the two groups before and after treatment.
| Indicator | Control group (n=36) | ALGS group (n=32) | P | ||
| Before treatment | After treatment | Before treatment | After treatment | ||
| Haematological parameters | |||||
| WBC (×109/L) | 11.89±1.74 | 11.81±1.87 | 11.76±1.66 | 12.23±0.18 | 0.208 |
| RBC (×1012/L) | 4.32±0.41 | 4.14±0.48 | 4.21±0.52 | 3.94±0.42 | 0.183 |
| PLT (×109/L) | 420±118.2 | 419.13±120.13 | 423±116.6 | 379.68±94.85 | 0.260 |
| Absolute value of Neu (×109/L) | 2.22±0.61 | 2.13±0.78 | 2.18±0.69 | 5.16±0.32 | <0.001 |
| Percentage of Neu (%) | 22.16±4.98 | 20.31±5.64 | 21.27±5.59 | 41.62±0.25 | <0.001 |
| Absolute value of Lym (×109/L) | 8.44±1.87 | 8.39±1.92 | 8.41±1.55 | 5.46±0.20 | <0.001 |
| Lym percentage (%) | 69.49±7.45 | 68.78±7.86 | 70.71±6.86 | 46.37±0.15 | <0.001 |
| Neu/Lym ratio | 0.23±0.12 | 0.24±0.03 | 0.23±0.16 | 0.97±0.06 | <0.001 |
| Absolute value of MO (×109/L) | 1.02±0.44 | 1.03±0.43 | 1.03±0.41 | 1.06±0.43 | 0.838 |
| Biochemical indices | |||||
| γ-GT (U/L) | 140.12±59.2 | 138.00±66.08 | 141.32±61.1 | 440.25±56.84 | <0.001 |
| DBil (μmol/L) | 73.44±28.16 | 70.00±34.96 | 72.28±33.26 | 116.88±54.82 | 0.009 |
| TBA (μmol/L) | 94.29±39.55 | 95.70±40.60 | 94.41±45.33 | 135.32±15.53 | <0.001 |
| GPT (U/L) | 156.42±80.22 | 161.75±79.15 | 159.11±78.71 | 234.29±47.87 | 0.001 |
| GOT (U/L) | 183.36±58.11 | 179.50±61.96 | 180.11±60.73 | 146.82±52.84 | 0.089 |
| TC (mmol/L) | 4.11±1.21 | 4.56±1.17 | 4.33±1.62 | 7.88±1.47 | <0.001 |
| TRIG (mmol/L) | 1.32±1.23 | 1.54±1.07 | 1.44±1.09 | 2.43±1.79 | 0.115 |
| LDL (mmol/L) | 2.66±0.89 | 2.53±0.91 | 2.60±1.01 | 4.25±1.09 | <0.001 |
| HDL (mmol/L) | 0.89±0.51 | 0.95±0.46 | 0.92±0.64 | 1.57±1.04 | 0.054 |
| TP (g/L) | 52.12±4.48 | 55.95±5.41 | 54.36±5.16 | 68.78±2.13 | <0.001 |
| Alb (g/L) | 40.18±4.15 | 42.83±3.77 | 41.62±3.89 | 44.56±2.31 | 0.072 |
| GLB (g/L) | 12.16±3.49 | 13.12±3.14 | 13.41±3.89 | 24.74±2.03 | <0.001 |
WBC, white blood cell; RBC, red blood cell; PLT, platelet; Neu, neutrophil; Lym, lymphocyte; MO, monocyte; γ-GT, γ-glutamyl transpeptidase; DBil, conjugated bilirubin; TBA, total bile acid; GPT, glutamic pyruvic transaminase; GOT, glutamic oxaloacetic transaminase; TC, total cholesterol; TG, triglyceride; LDL, low density lipoprotein; HDL, high density lipoprotein; TP, total protein; Alb, albumin; GLB, globulin. Control group: children with unexplained cholestasis; ALGS group: children with neonatal intrahepatic cholestasis caused by Alagille syndrome.
most common eye dysfunction, and the incidence rate can be as high as 95% (Andersson et al., 2017; . This study found that the incidences of cardiovascular malformation, special face, vertebral body changes and eye abnormalities in neonatal intrahepatic cholestasis caused by ALGS were 74.19%, 77.42%, 51.61% and 51.61%, respectively, which were raised than those in neonatal intrahepatic cholestasis caused by Citrin deficiency. It is suggested that the above manifestations can be used as important manifestations in the diagnosis of ALGS.
For patients whose clinical manifestations are not obvious, blood routine and laboratory biochemical tests will also provide critical clues for the cholestasis diagnosis . In this study, neutrophils, lymphocytes, and their ratio play critical role in the diagnosis of intrahepatic cholestasis caused by ALGS. Immune cells are a group of cells that interact with each other. Lymphocytes and neutrophils are both important cellular components of the body’s immune response function. Among them, neutrophils are the most important components of leukocytes in the blood. They are rich in lysosomes, can release a variety of inflammatory factors, and playcritical role in the body’s non-specific immunity. It is found that when intrahepatic cholestasis occurs, the proportion of lymphocyte subsets is out of balance and the immune function is significantly decreased, thus aggravating the patient’s condition . The levels of neutrophils and lymphocytes play critical role in reflecting the progression of liver disease. Among biochemical indices, γ-GT is most expressed in the liver during the embryonic stage, and is an important index to reflect the severity of the disease . The normal secretion and fluidity of bile mainly depends on the ATP-dependent pumps and secretory proteins of hepatocytes and bile duct epithelial cells. However, Jag1 gene mutations in patients with ALGS can cause functional impairment of small bile ducts, which can easily lead to cholestasis, while cholestasis and poisoning can damage hepatocytes, lead to damage to liver repair ability, and then aggravate cholestasis and enter a vicious circle . TP and Alb which reflect the state of liver disease and nutritional status of the body will be significantly decreased of the body is malnourished . The results show that biochemical indexes such as γ-GT, conjugated bilirubin, TBA, TP and GLB play critical role over time in distinguishing intrahepatic cholestasis caused by ALGS.
Declarations
Acknowledgments
Thanks are due to the members from Wuhan Children’s Hospital. They collected samples, obtained data, and provided theoretical guidance.
Ethical approval
The study was carried out in compliance with guidelines issued by ethical review board committee of Wuhan Children’s Hospital, China. The official letter would be available on fair request to corresponding author.
IRB approval
This study was approved by the Advanced Studies Research Board of Wuhan Children’s Hospital, Hubei Province, China.
Ethical statement
Ethics Committee approval was obtained from the Institutional Ethics Committee of “Wuhan Children’s Hospital” to the commencement of the study.
Supplementary material
There is supplementary material associated with this article. Access the material online at: https://dx.doi.org/10.17582/journal.pjz/20240114052214
Statement of conflict of interest
The authors have declared no conflict of interest.
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