INTRODUCTION
Morus of the family Moraceae is a small plant genus having 19 species worldwide [1]. Economically important species are Morus alba (white mulberry), Morus indica (MI, Indian mulberry), Morus nigra (MN, black mulberry), and Morus rubra (red mulberry). China has 11 Morus species of which 5 are endemic and 1 is introduced [2].
Morus alba L., comprising M. alba var. alba and M. alba var. multicaulis, is a fast-growing monoecious and deciduous shrub or medium-sized tree without buttress [2−5]. Young plants produce multiple stems via coppicing. The bark is brownish–gray with vertical fissures, lenticels, and white or cream-colored latex. Leaves of M. alba (MA) are glossy green, alternately arranged, cordate at the base, and acuminate at the apex. Leaf margins are serrated, leaf petioles are long and slender, and leaf blades vary from unlobed to almost palmate. Fruits are drupes or sorosis that are white when young, turning reddish when mature, and black when ripe [2−5]. Leaves, twigs, and maturing fruits of MA are shown in Figure 1.
The whole plant, leaf, fruit, twig, and root of MA have medicinal values. Chemical constituents comprise steroids, tannins, phytosterols, glycosides, alkaloids, carbohydrates, proteins, and amino acids, as well as saponins, triterpenes, phenols, flavonoids, benzofurans, anthocyanins, polysaccharides, anthraquinones, and glycosides [5−7]. Pharmacological properties include antioxidant, antimicrobial, anti-inflammatory, anti-diabetic, hypolipidemic, anti-obesity, anti-atherosclerotic, neuroprotective, hepatoprotective, tyrosinase inhibitory, and cardioprotective [5−7]. The beneficial effects of MA leave against cardiometabolic risks have been reviewed [8]. The chemical constituents, medicinal properties, clinical trials, and patents of twigs of MA (Ramulus Mori) have recently been reviewed [9].
In this review, the clinical studies of Morus species (mostly MA) to date are briefly described. These studies are categorized as anti-diabetic properties (2007−2022) and other pharmacological properties (2001−2021) in chronological order. Relevant to the findings of these studies are mention of compounds, their classes, and bioactivities.
CLINICAL STUDIES
Anti-diabetic properties
There are 23 clinical studies on anti-diabetic properties of Morus species (Table 1). Five studies were undertaken in Japan; three studies each in the USA, Korea, China, and India; and two studies each in Iran and the UK. Single studies were conducted in Thailand and Poland. All studies were on MA except one study on MN. Plant parts of MA clinically tested were mostly leaves with fruits and twigs lesser studied.
OTHER PHARMACOLOGICAL PROPERTIESS
There are nine clinical studies on other pharmacological properties of Morus species (Table 2). Four studies were undertaken in Thailand, two studies in China, and single studies were conducted in Japan, India, and Brazil. One study was tested on MN and MI each. Clinical studies on other pharmacological properties of Morus species include hypolipidemic (3), cognitive enhancement (2), coronary heart disease (CHD) attenuation (2), anti-obesity (1), and climacteric improvement (1).
 | Figure 1. Leaves (left), twigs (middle), and maturing fruits (right) of Morus alba. [Click here to view] |
BIOACTIVE COMPOUNDS
Anti-diabetic
Compounds in MA leaves, fruits, and twigs with anti-diabetic activities include 1-deoxynojirimycin (DNJ), quercetin, dihydroquercetin kaempferol, rutin, chlorogenic acid chalcomoracin, morachalcone, and isobavachalcone [6,42,43]. Steppogenin-4’-O-β-D-glucoside and mulberroside A from the root bark of MA significantly reduced the fasting blood glucose level in alloxan-induced diabetic mice [44]. Rutin and quercetin-3-O-β-D-glucoside, two anti-diabetic flavonoids from the fruit of MA, improved glucose uptake in 3T3-L1 adipocytes [45]. The mechanism involved Akt-mediated insulin signaling pathway or AMP-activated protein kinase activation. A high-purity polysaccharide from mulberry leaf extract (MLE) (99.8% purity) exhibited anti-diabetic effects in streptozotocin-induced diabetic rats with effects equivalent to glibenclamide (GBC), an anti-diabetic drug [46].
 | Table 1. Clinical studies on anti-diabetic properties of Morus species. [Click here to view] |
DNJ, an alkaloid from MA leaves and twigs, is a potent α-glucosidase inhibitor and is effective in suppressing high blood glucose levels in human subjects, thus preventing T2D [4,9]. In diabetic mice, DNJ significantly decreased serum glucose and insulin levels, improved serum lipid contents, and reversed insulin resistance [47]. DNJ prevents the secretion of insulin and thus lowers fasting and post-prandial blood glucose levels associated with T2D [48]. Out of 21 clinical studies on anti-diabetic activities, 12 studies have been attributed to DNJ (Table 1).
 | Table 2. Clinical studies on other pharmacological properties of Morus species. [Click here to view] |
Hypolipidemic
Mulberroside A was a stilbenoid isolated and purified from the ethanol root extract of MA while oxyresveratrol was produced from enzymatic conversion. Both compounds exhibited hypolipidemic effects in rats on a high-cholesterol diet. Rats orally treated with mulberroside A and oxyresveratrol significantly decreased serum lipids, coronary artery risk index, and atherogenic index [49]. From the leaf of MA, a benzofuran derivative, a flavonoid, and an alkaloid displayed potent lipolytic activity in 3T3-L1 cells with values from 15.4% to 21.2% [43]. Rutin and quercetin-3-O-β-D-glucoside, two anti-diabetic flavonoids from the fruit of MA, reduced lipid accumulation in adipocytes [45].
Anti-obesity
The ability of MLE to suppress obesity and reduce visceral adipose tissues has been attributed to polyphenols such as quercetin, kaempferol, rutin, caffeic acid, and chlorogenic acid [6]. From MLE, 2’,7-dihydroxy-4’-methoxy-8-prenylflavan (flavan), isobavachalcone, and morachalcone B (chalcones) inhibited 3T3-L1 preadipocytes with IC50 values of 37, 43, and 48 μM, respectively [50]. A pectic polysaccharide from MFE, named JS-MP-1, displayed an anti-obesity effect by inhibiting pre-adipocytes via reducing fat cells and adipose tissue [51].
Cognitive enhancement
Mulberrofuran G (2.13 and 9.72 μM) and albanol B (2.47 and 1.39 μM) from the root bark of MA possessed strong acetylcholinesterase and butyrylcholinesterase inhibitory activities, respectively [52]. These activities showed their ability to treat cognitive dysfunction associated with Alzheimer’s disease (AD). Among four moracins isolated from the root of MA, the inhibition of BACE1, a beta-secretase enzyme in AD, moracin S was the strongest [53].
CHD attenuation
CHD is a common cardiovascular disease that causes human disability and death [54]. Among the various symptoms of CHD are angina pectoris, blood stasis syndrome, and atherosclerosis [38,39]. The underlying mechanisms of CHD are associated with inflammatory stress responses [55]. Compounds in Morus species that possess anti-inflammatory properties include DNJ and oxyresveratrol. DNJ, the main component in MA alkaloid tablets, possessed anti-inflammatory properties [56]. The alkaloid checked inflammation via regulation of mitogen-activated kinase signaling. DNJ markedly down-regulated interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) cytokine levels in lipopolysaccharide (LPS)-induced RAW 264.7 and bone marrow-derived macrophage cells. Oxyresveratrol, a stilbenoid, is another ingredient of MA that exerts anti-inflammatory activity via inhibition of leukocyte migration, and involvement of mitogen-activated ERK (MEK)/extracellular signal-regulated kinase (ERK) signaling [57]. Oxyresveratrol from MA also inhibited LPS-induced translocation of nuclear kappa B and cyclooxygenase-2 activity in RAW 264.7 cells [58,59]. The anti-inflammatory activity of oxyresveratrol has also been reported in RAW 264.7 cells, Jurkat leukemic T cells, and C28/I2 chondrocytes [60]. Quercetin 3-(6-malonylglucoside), a flavonol from MLE, attenuated atherosclerosis in low-density lipoprotein (LDL) receptor-deficient mice [61]. Among 36 compounds from the twig of MA, albanin D and 3-methyl-1-phenyl-1,3-butadiene exhibited the strongest anti-inflammatory activity of 4.1 and 2.2 μM by inhibiting NO production in RAW 264.7 cells [62]. The potent anti-inflammatory activity of prenylated flavonoids from the root of MA and MN has been reported [63]. Noteworthy is kuwanon C with an IC50 value of 1.7 μM. Albanol, an arylbenzofuran derivative from the root bark of MA, had the strongest anti-inflammatory effects towards RAW 264.7 cells, followed by sanggenon B and sanggenon D [64].
CONCLUSION
Objectives of clinical studies on MA include the effective dosage, duration, timing, and administration. Materials used include mulberry tea, MLE powder, and confections containing enriched compounds such as DNJ, and standardized extract, e.g., reducose. Subjects are children, middle-aged adults, and elderly people, including people with CHD, impaired glucose tolerance (IGT), Pompe disease (PD), dyslipidemia, and climacteric symptoms. Some clinical studies on MA are designed to compare diabetic patients, people with dyslipidemia, and healthy or nondiabetic volunteers. Diabetic patients include post-prandial pre-diabetic, and borderline subjects. Placebo groups serve as controls. Further research is needed on the anti-diabetic mechanisms of mulberry leaves at the molecular level, which may involve multiple pathways. While most clinical trials have shown that mulberry leaves regulate blood glucose and lipid metabolism, research focusing on the safety of mulberry leaves is lacking. Studies are therefore needed to understand the distribution, absorption, metabolism, and excretion of mulberry compounds.
AUTHOR CONTRIBUTIONS
The author made substantial contributions to the conception and design, acquisition of data, or analysis and interpretation of data; took part in drafting the article or revising it critically for important intellectual content; agreed to submit to the current journal; gave final approval of the version to be published; and agree to be accountable for all aspects of the work. The author is eligible to be an author as per the International Committee of Medical Journal Editors (ICMJEs) requirements/guidelines.
FINANCIAL SUPPORT
Assoc. Prof. Eric W. C. Chan, the Lead and Sole Author, acknowledges that the funds for the publication of this review in the Journal of Applied Pharmaceutical Science (JAPS) as article processing charges (APCs) are provided by UCSI University. The author is grateful for the World’s Top 2% Scientist Research Grant, awarded by CERVIE (Grant Code: T2S-2023/004).
CONFLICTS OF INTEREST
The authors report no financial or any other conflicts of interest in this work.
ETHICAL APPROVALS
This study does not involve experiments on animals or human subjects.
DATA AVAILABILITY
All data generated and analyzed are included in this research article.
USE OF ARTIFICIAL INTELLIGENCE (AI)-ASSISTED TECHNOLOGY
The authors declares that they have not used artificial intelligence (AI)-tools for writing and editing of the manuscript, and no images were manipulated using AI.
PUBLISHER’S NOTE
All claims expressed in this article are solely those of the authors and do not necessarily represent those of the publisher, the editors and the reviewers. This journal remains neutral with regard to jurisdictional claims in published institutional affiliation.
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