Identification of Steroid Alkaloids in Resveratrol and Study on Their Promoting Lysosomal Activity
The dysfunction of lysosomal biosynthesis is closely related to neurodegenerative diseases. Lysosomes, as the main organelles responsible for degradation in cells, help to clear damaged and aging organelles and misfolded proteins in cells. They are crucial for maintaining the homeostasis of the nervous system and are closely related to neurodegenerative diseases. Resveratrol is a perennial herbaceous plant of the genus Resveratrol in the family Liliaceae. There are about 40 species of plants in the genus Resveratrol worldwide, with 13 species and 1 variety recorded in China and 4 species in Yunnan. In Ge Hong’s book “Emergency Prescriptions for Elbow Reserve” (1955 edition), it is recorded that resveratrol can treat “stroke patients who are unconscious and have clenched teeth”; The Chinese Pharmacopoeia of Traditional Chinese Medicine also refers to the use of Chinese medicine resveratrol for the treatment of stroke aphasia, headache, jaundice, scabies, malaria, dysentery, as well as for insecticidal and emetic purposes. The efficacy of resveratrol is related to neurodegenerative diseases. So far, steroidal alkaloids in Veratrum are considered to be the active substance basis of plants in the genus Veratrum. More than 200 species have been isolated, which can be divided into four types of skeletons: solanidine, vera ramine, jervine, and cevanine (see Figure 1). Most of them have C-27 steroid skeletons, among which solanidine and vera ramine belong to the cholestane alkaloids. The A, B, and C rings are all six membered rings, and the D ring is a five membered ring. According to the different nitrogen atom formation, E/F forms an indole ring with N at the C-16 position, which is a solanidine alkaloid. Veraramine alkaloids mostly do not have an E ring, and the F ring is a pyridine ring; Jervine and cevanine belong to the class of isocholestane alkaloids, characterized by a c-nor-homo [14 (13 → 12) – abeo] ring system. A. The B and D rings are hexagonal rings, while the C ring is a pentagonal ring. The cevanine alkaloids form a new hexagonal ring E by combining the N atom of the C-17 and F rings. The difference between the skeleton of jervine and cevine alkaloids lies in the presence of a covalent bond between C-17 and the nitrogen atom. Current studies have shown that jervine alkaloids have strong analgesic activity; Veratramine type alkaloids have a wide range of activities and are 5-HT receptor antagonists. They participate in the regulation of AP-1 target DNA sequences and have the ability to block sodium channels. Veratramine type alkaloids also have strong in vivo and in vitro platelet aggregation inhibitory activity, inhibiting arterial thrombosis, and have been extensively studied in anti-tumor studies; Cyclopamine is the first Hedgehog (Hh) signaling pathway inhibitor that inhibits tumor growth caused by abnormal activation of this pathway. In summary, there have been no reports on the relationship between such alkaloids and neurodegenerative diseases to date.
The efficacy inspiration of resveratrol focuses on “stroke aphasia”, which is related to neurodegenerative diseases. Taking this as an opportunity, we chose the medicinal plant source of resveratrol – hairy leaf resveratrol – as the object of research. We studied the steroidal alkaloids in methanol extracts of the roots and rhizomes of Atractylodes macrocephala, with the aim of isolating chemical components that promote lysosome production from the roots and rhizomes of Atractylodes macrocephala, elucidating the active substance basis that affects lysosome function in Atractylodes macrocephala, and providing lead compounds for neurodegenerative diseases.

Italian scholar Ballabio’s research group found that lysosome generation is mainly regulated at the transcriptional level by transcription factors TFEB and TFE3 of the MiTF/TFE family, which are expressed in various tissues. Under normal cellular conditions, TFEB is phosphorylated and binds to 14-3-3 protein, dispersing throughout the cytoplasm. However, under external stimuli such as hunger or impaired lysosomal function, or when cells are subjected to stress, TFEB undergoes dephosphorylation and enters the nucleus, regulating the expression of downstream genes and widely participating in various physiological and pathological activities. The phosphorylation of TFEB is also regulated by various protein molecules both in vivo and in vitro. The TFEB/CLEAR network serves as a regulatory center to promote lysosome generation and related lysosome degradation functions (macromolecular degradation, autophagy lysosome pathway, mitosis, and lipophagy, etc.), playing a crucial role in improving neurodegenerative diseases. At present, literature reports on small molecule compounds that regulate TFEB nuclear entry include giant diterpenes HEP14 and HEP15, saponins XVII from Polygonatum sibiricum, and curcumin derivatives; No reports have been found on the promotion of lysosome formation by steroid alkaloids. Therefore, we are searching for lead compounds with new structural types that promote lysosomal biogenesis and exploring their mechanisms of action, providing new ideas for the development of new therapeutic drugs for neurodegenerative diseases.
Starting from the guiding principles of traditional Chinese medicine theory, this study focuses on the treatment of “stroke aphasia” with resveratrol, and speculates that it is related to neurodegenerative diseases. Taking this as an opportunity, research on the chemical composition of hairy leaf resveratrol is carried out, and the active substance basis of resveratrol is deeply explored to explore whether it can promote lysosome generation and provide lead compounds for the treatment of neurodegenerative diseases. In this experiment, 8 steroid alkaloids were isolated from the leaves of the Chinese parasol tree. Compounds 1, 2, 3, 6, and 7 can all promote lysosome formation, with compound 6 showing the best effect. We found that compound 6 can promote the transcription factor TFEB into the nucleus. In the future, we will verify the lysosome function and study the mechanism of action of the screened compounds.