Traditional Chinese Medicine To Treat Oligoasthenozoospermia By Regulating Cellular Autophagy Ⅱ

3. The Role of Autophagy in Sperm Maturation

To acquire fertilization capability, elongated spermatids must undergo a maturation process, which occurs primarily in the epididymis. First, sperm are transported from the seminiferous tubules through the rete testis into the caput epididymis, propelled by flagellar motion and smooth muscle contraction, and gradually gain motility and fertilization capacity in the cauda epididymis [64].

During ejaculation and female reproductive tract transit, sperm undergo a series of biochemical modulations, including phosphorylation, alkalinization, and membrane hyperpolarization, facilitated by cystic fibrosis transmembrane conductance regulators (CFTR) and protein kinase A (PKA), enabling penetration of the oocyte cumulus and zona pellucida [65].

One crucial factor in this process is reactive oxygen species (ROS). While excessive ROS can be detrimental to sperm function, physiological levels of ROS are essential for capacitation and acrosomal reaction. Therefore, the balance between ROS and autophagy is vital for sperm maturation.

ROS modulate autophagy through transcriptional and post-transcriptional pathways, including:

ROS–FOXO3–LC3/BNIP3 axis

ROS–Nrf2–p62–autophagy

ROS–HIF1–BNIP3/NIX pathway

ROS–TIGAR–autophagy [66]

Conversely, autophagy regulates intracellular ROS via chaperone-mediated autophagy (CMA), mitotic signaling, and p62 cargo pathways [66].

Recent studies have shown that in leucine-treated zebrafish, autophagy modulation through the PI3K/Akt pathway can impair autophagosome-lysosome fusion, thereby affecting sperm motility [67].

 

Click The Pic To Get More About TCM Herb For Improving Sperm Motility

 

 

4. Progress in Traditional Chinese Medicine (TCM) for Regulating Autophagy in the Treatment of Oligoasthenozoospermia (OAS)

4.1 TCM Theory and Pathogenesis of OAS

In TCM theory, asthenozoospermia (reduced motility) is often attributed to "yang deficiency unable to transform qi", while oligospermia (low sperm count) is associated with "yin deficiency failing to form essence". Hence, yang invigorating (to warm and activate qi) and yin nourishing (to promote essence formation) therapies are commonly employed to restore sperm vitality and quantity [68–70].

This traditional understanding aligns with modern findings that autophagy plays a dual role in spermatogenesis and sperm maturation. Thus, TCM may act at both macro (yin-yang regulation) and micro (autophagy pathway modulation) levels to enhance male fertility.

4.2 Mechanistic Insights: TCM and Autophagy Regulation

Recent research has linked TCM's therapeutic effects on OAS to several molecular mechanisms, including:

Epigenetic regulation (DNA methylation, histone modification, non-coding RNAs) [71–73]

Mitochondrial apoptosis and oxidative stress

Autophagy-related signaling, such as:

PI3K/Akt/mTOR

AMPK/ULK1

MEK5/ERK5

PINK1/Parkin

Nrf2/p62 axis [74–79]

These findings highlight TCM's multi-component, multi-target, and multi-pathway mechanisms in treating complex diseases, including OAS.

4.3 Classic TCM Formulas

Wuzi Yanzong Pill has been shown to:

Upregulate LC3, LAMP-2, and Atg5 in Sertoli cells

Improve autophagy balance via AKT/mTORC1–mTORC2 modulation

Enhance sperm concentration, motility, and testicular histology in aging or damaged models [80–82]

Guilu Erxian Gel and TCM-medicated serum:

Activate the Keap1/Nrf2 pathway

Inhibit oxidative stress-induced autophagy

Improve spermatogonial cell viability by modulating LC3B, p62, and mTOR expression [83]

 

4.4 TCM-Based Prescriptions

Jingui Yulin Formula:

Regulates the PINK1/Parkin pathway

Reduces abnormal autophagy and improves mitochondrial membrane potential and ATP levels in sperm [85]

Qixiong Formula:

Modulates PI3K/Akt/mTOR signaling

Decreases LC3-II/LC3-I ratio, Beclin-1, and increases p62 expression, enhancing sperm motility [86]

Sancai Lianmei Granules:

Activates Nrf2/HO-1

Reduces oxidative stress and autophagic vesicle formation, improving spermatogenesis in diabetic models [87]

4.5 TCM Monomers and Active Ingredients

Jiangsu oyster polysaccharide, celery seed oil, purslane extract, cordycepin, ginseng saponins, and Ganoderma lucidum spore powder have all been shown to:

Regulate autophagy markers (LC3, Beclin-1, Atg5, Atg7)

Modulate oxidative stress parameters (SOD, MDA, CAT, GSH)

Improve sperm quality and testicular structure by targeting various pathways including Nrf2/HO-1, PI3K/Akt, LKB1/AMPK/ULK1, and PTEN/E3 [88–97]

 

Table 1. Mechanisms of Traditional Chinese Medicine (TCM) Compound or Monomer Acting on Oligoasthenozoospermia via Regulation of Autophagy

TCM Formula / Monomer	Research Model	Mechanism of Action
Wuzi Yanzong Pill [80]	Cyclophosphamide-induced OAS in male SD rats	↑ Autophagy, ↑ LC3, ↑ LAMP-2
Wuzi Yanzong Pill [81]	Naturally aging male C57BL/6 mice	↑ LC3B, ↑ Atg5, ↓ p62, ↓ p-AKT, ↓ p-mTORC1, ↑ mTORC2
Wuzi Yanzong Pill-containing serum [82]	TM4 Sertoli cells under oxidative stress	↓ LC3, ↓ Beclin-1
Guilu Erxian Gel-containing serum [83]	GC-1 spermatogonia treated with H₂O₂ and siRNA	↓ Autophagy via ↑ Keap1/Nrf2 pathway, ↑ p62, ↑ p-mTOR, ↓ LC3B-II/LC3B-I
Wuzi Yanzong Pill [84]	SD rat model of reproductive aging	Not specified (inferred involvement of autophagy)
Jingui Yulin Formula [85]	SD rats with OAS induced by ornidazole	↓ Autophagy via ↓ Pink1/Parkin pathway, ↓ LC3B-II/LC3B-I, ↓ Beclin-1, ↑ TOM20, ↑ p62
Qixiong Formula [86]	SD rats with OAS induced by ornidazole	↓ Autophagy via ↓ Beclin-1, ↓ LC3-II/LC3B-I, ↑ p62, ↑ p-PI3K, ↑ p-AKT, ↑ p-mTOR
Sancai Lianmei Granule [87]	C57BL/6 mice with diabetes-induced testicular damage	Activates Nrf2/HO-1 pathway, ↓ LC3, ↓ p62, ↓ Beclin-1
Yijing Formula-containing serum [88]	Oxidative stress-induced TM3 Leydig cells	↓ LC3B, ↓ Beclin-1, ↑ p-PI3K/p-AKT, ↑ Bcl-2, ↑ SOD, ↑ T-AOC
Qiangjing Tablet [89]	OAS rat model induced by metronidazole	↑ PHB, ↑ Beclin-1, ↑ LC3-II, ↑ ROS, ↑ LKB1, ↑ AMPKα, ↑ ULK1
Huangjing Zanyu Capsule [90]	OAS rat model induced by cyclophosphamide and GC-2 cell line model	↓ Autophagy via ↑ Akt/mTOR, ↓ LC3, ↓ Beclin-1, ↑ p-AKT, ↑ p-mTOR
Jiangsu Oyster Polysaccharide [91]	BALB/c mice and TM4 cells under oxidative stress	↓ LC3B-II/LC3B-I, ↓ Beclin-1, ↑ SOD, ↑ GSH-Px, ↓ ROS
Apigenin [92]	SD rats with arsenic-induced testicular damage	↓ LC3A, ↓ LC3B, ↓ MAPK-14, ↑ SOD, ↑ CAT, ↑ GSH-Px, ↑ GSH
Portulaca oleracea extract + Chicory water extract [93]	SD rats with dexamethasone-induced testicular dysfunction	↓ LC3B-II/LC3B-I, ↓ Atg5, ↓ Atg7, ↑ p62, ↑ p-mTOR
Cardamonin [94]	Diabetic rats with hyperglycemia-induced testicular dysfunction	↑ Nrf2 via p62-dependent autophagy, ↓ Keap1
Hesperidin [95]	SD rats with colistin-induced reproductive toxicity	↑ LC3B-II/LC3B-I, ↓ MDA, ↑ SOD, ↑ CAT, ↑ GSH, ↑ GSH-Px
Cordycepin [96]	Naturally aging rats with testicular dysfunction	↑ AR, ↑ FSHR, ↑ LHR, ↑ CREB-1, ↑ Nectin-2, ↑ Inhibin-α, ↑ mTOR, ↓ SIRT1
Ginsenoside from Polygonatum [97]	High-fat-diet-induced testicular injury in C57BL/6 mice	↑ ZO-1, ↑ Occludin, ↑ N-cadherin, ↑ E-cadherin, ↑ β-catenin, ↓ LC3B, ↑ p-AKT, ↑ p-mTOR
Ganoderma lucidum spore powder [74]	STZ-induced diabetic testicular injury in SD rats	↑ LC3B-II/LC3B-I, ↑ PTEN, ↑ E3, ↓ apoptosis

 

5. Summary and Perspectives on Traditional Chinese Medicine Regulation of Autophagy in the Treatment of Oligoasthenozoospermia (OAS)

In summary, autophagy plays a bidirectional regulatory role throughout the processes of spermatogenesis and sperm maturation. Traditional Chinese Medicine (TCM) may exert synergistic therapeutic effects on OAS by modulating autophagy-related signaling pathways, including PI3K/Akt/mTOR, AMPK, and PINK1/Parkin, as well as key autophagy markers.

Therefore, autophagy and its associated molecular markers and signaling pathways may represent promising therapeutic targets for OAS. However, current research still faces several limitations:

 

(1) Unclear Identification of Active Compounds and Targets

Due to the complex multi-component nature of TCM, it remains unclear which specific herbs or chemical constituents within classic formulas or self-made prescriptions are responsible for targeting autophagy in OAS. Moreover, most existing studies focus on compound formulas, self-prepared prescriptions, or individual monomers, with limited investigation into single-herb effects, which may hinder the full exploitation of TCM's unique advantages.

✅ Future direction: Leverage systems pharmacology, bioinformatics, high-throughput mass spectrometry, and multi-omics technologies to identify bioactive components and elucidate their mechanisms of action in regulating autophagy for OAS treatment.

 

(2) Lack of High-Quality Clinical Evidence

Most studies to date have been conducted at the in vitro and animal model levels. There is a pressing need for well-designed, large-scale clinical trials to provide high-level evidence for the efficacy and safety of TCM in regulating autophagy for the treatment of OAS.

✅ Future direction: Conduct randomized controlled trials (RCTs) and multi-center clinical studies to support the clinical translation of TCM therapies.

 

(3) Limited Scope of Autophagy Types Studied

Current research primarily focuses on macroautophagy, while other forms such as microautophagy, chaperone-mediated autophagy (CMA), and selective autophagy (e.g., mitophagy, reticulophagy, ferroptosis-related autophagy) are rarely explored.

✅ Future direction: Expand investigations into diverse subtypes of autophagy and their roles in the pathogenesis and treatment of OAS.

 

(4) Lack of Syndrome-Differentiated Animal Models in TCM Research

There is a lack of syndrome-based animal models that reflect TCM diagnostic categories (e.g., kidney yang deficiency, damp-heat, qi stagnation). Moreover, research has yet to fully integrate syndrome differentiation (辨证论治) theory with autophagy-related mechanisms.

✅ Future direction: Develop TCM pattern-based OAS models, and integrate syndrome differentiation with autophagy biology to guide targeted TCM interventions.

 

Conclusion and Outlook

Overall, TCM presents a promising and holistic approach to treating OAS by modulating autophagy and restoring reproductive function. Future research should focus on identifying active compounds, validating clinical efficacy, exploring non-canonical autophagy pathways, and establishing syndrome-specific models, thereby expanding therapeutic strategies and improving the clinical outcomes for patients with OAS.