是的，在Windows中，您可以通过以下方式执行此操作：

import ctypes
from ctypes import wintypes

path = r"C:\temp\test.txt"

# -----------------------------------------------------------------------------
# generic strings and constants
# -----------------------------------------------------------------------------

ntdll = ctypes.WinDLL('ntdll')
kernel32 = ctypes.WinDLL('kernel32', use_last_error=True)

NTSTATUS = wintypes.LONG

INVALID_HANDLE_VALUE = wintypes.HANDLE(-1).value
FILE_READ_ATTRIBUTES = 0x80
FILE_SHARE_READ = 1
OPEN_EXISTING = 3
FILE_FLAG_BACKUP_SEMANTICS = 0x02000000

FILE_INFORMATION_CLASS = wintypes.ULONG
FileProcessIdsUsingFileInformation = 47

LPSECURITY_ATTRIBUTES = wintypes.LPVOID
ULONG_PTR = wintypes.WPARAM

# -----------------------------------------------------------------------------
# create handle on concerned file with dwDesiredAccess == FILE_READ_ATTRIBUTES
# -----------------------------------------------------------------------------

kernel32.CreateFileW.restype = wintypes.HANDLE
kernel32.CreateFileW.argtypes = (
    wintypes.LPCWSTR,      # In     lpFileName
    wintypes.DWORD,        # In     dwDesiredAccess
    wintypes.DWORD,        # In     dwShareMode
    LPSECURITY_ATTRIBUTES,  # In_opt lpSecurityAttributes
    wintypes.DWORD,        # In     dwCreationDisposition
    wintypes.DWORD,        # In     dwFlagsAndAttributes
    wintypes.HANDLE)       # In_opt hTemplateFile
hFile = kernel32.CreateFileW(
    path, FILE_READ_ATTRIBUTES, FILE_SHARE_READ, None, OPEN_EXISTING,
    FILE_FLAG_BACKUP_SEMANTICS, None)
if hFile == INVALID_HANDLE_VALUE:
    raise ctypes.WinError(ctypes.get_last_error())

# -----------------------------------------------------------------------------
# prepare data types for system call
# -----------------------------------------------------------------------------

class IO_STATUS_BLOCK(ctypes.Structure):
    class _STATUS(ctypes.Union):
        _fields_ = (('Status', NTSTATUS),
                    ('Pointer', wintypes.LPVOID))
    _anonymous_ = '_Status',
    _fields_ = (('_Status', _STATUS),
                ('Information', ULONG_PTR))

iosb = IO_STATUS_BLOCK()

class FILE_PROCESS_IDS_USING_FILE_INFORMATION(ctypes.Structure):
    _fields_ = (('NumberOfProcessIdsInList', wintypes.LARGE_INTEGER),
                ('ProcessIdList', wintypes.LARGE_INTEGER * 64))

info = FILE_PROCESS_IDS_USING_FILE_INFORMATION()

PIO_STATUS_BLOCK = ctypes.POINTER(IO_STATUS_BLOCK)
ntdll.NtQueryInformationFile.restype = NTSTATUS
ntdll.NtQueryInformationFile.argtypes = (
    wintypes.HANDLE,        # In  FileHandle
    PIO_STATUS_BLOCK,       # Out IoStatusBlock
    wintypes.LPVOID,        # Out FileInformation
    wintypes.ULONG,         # In  Length
    FILE_INFORMATION_CLASS)  # In  FileInformationClass

# -----------------------------------------------------------------------------
# system call to retrieve list of PIDs currently using the file
# -----------------------------------------------------------------------------
status = ntdll.NtQueryInformationFile(hFile, ctypes.byref(iosb),
                                      ctypes.byref(info),
                                      ctypes.sizeof(info),
                                      FileProcessIdsUsingFileInformation)
pidList = info.ProcessIdList[0:info.NumberOfProcessIdsInList]
print(pidList)